- ￥100 - 1000
- Xybio
- 上海
- P2504
- 2025年07月13日
9 年钻石会员
企业认证
相关产品推荐更多 >
万千商家帮你免费找货
0 人在求购买到急需产品
- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20
- 保质期：
2年
- 英文名：
Plasmid#79873;pAX01-dCas9
- 库存：
100
- 供应商：
上海烜雅生物科技有限公司
- 规格：
干粉/液体
基本信息
名称：pJMP1枯草编辑质粒
别称: Plasmid#79873;pAX01-dCas9
质粒属性
质粒简介
pJMP1质粒是一个枯草杆菌基因编辑质粒。
质粒图谱
质粒序列
LOCUS Exported 12080 bp ds-DNA circular SYN 14-MAY-2018
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..12080
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 1087..1278
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 1380..1520
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(1706..2294)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2465..3325)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3326..3430)
/gene="bla"
/note="AmpR promoter"
CDS complement(3541..3948)
/codon_start=1
/note="'lacA"
/translation="MKDYATVIDVKTASVEAVYQEDFYARTPAVTSHEYQQGKAYFIGA
RLEDQFQRDFYEGLITDLSLSPVFPVRHGKGVSVQARQDQDNDYIFVMNFTEEKQLVTF
DQSVKDIMTGDILSGDLTMEKYEVRIVVNTH"
terminator 4074..4108
/note="lambda t0 terminator"
/note="minimal transcription terminator from phage lambda
(Scholtissek and Grosse, 1987)"
CDS complement(4140..5264)
/codon_start=1
/note="bme xylR"
/translation="MNQKLILDEILKNSPVSRATLSEITGLNKSTVSSQVNTLLEKDFI
FEIGAGQSRGGRRPVMLVFNKNAGYSIGIDIGVDYLNGILTDLEGNIILEKTSDLSSSS
ASEVKEILFALIHGFVTHMPESPYGLVGIGICVPGLVDRHQQIIFMPNLNWNIKDLQFL
IESEFNVPVFVENEANAGAYGEKVFGMTKNYENIVYISINIGIGTGLVINNELYKGVQG
FSGEMGHMTIDFNGPKCSCGNRGCWELYASEKALLASLSKEEKNISRKEIVERANKNDV
EMLNALQNFGFYIGIGLTNILNTFDIEAVILRNHIIESHPIVLNTIKNEVSSRVHSHLD
NKCELLPSSLGKNAPALGAVSIVIDSFLSVTPIS"
CDS 5580..9686
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/note="dCas9"
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the D10A mutation in the RuvC
catalytic domain and the H840A mutation in the HNH
catalytic domain"
/translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNL-SDAILLS-DILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLS-DYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
terminator 9710..9781
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 9797..9824
/note="T7Te terminator"
/note="phage T7 early transcription terminator"
promoter complement(9899..10003)
/gene="bla"
/note="AmpR promoter"
CDS 10382..11119
/codon_start=1
/note="Erm"
/translation="MNKNIKYSQNFLTSEKVLNQIIKQLNLKETDTVYEIGTGKGHLTT
KLAKISKQVTSIELDSHLFNLSSEKLKLNTRVTLIHQDILQFQFPNKQRYKIVGSIPYH
LSTQIIKKVVFESHASDIYLIVEEGFYKRTLDIHRTLGLLLHTQVSIQQLLKLPAECFH
PKPKVNSVLIKLTRHTTDVPDKYWKLYTYFVSKWVNREYRQLFTKNQFHQAMKHAKVNN
LSTVTYEQVLSIFNSYLLFNGRK"
terminator 11289..11375
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 11394..11421
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 11501..11535
/note="lambda t0 terminator"
/note="minimal transcription terminator from phage lambda
(Scholtissek and Grosse, 1987)"
CDS complement(11559..12074)
/codon_start=1
/note="lacA'"
/translation="MSKLEKTHVTKAKFMLHGGDYNPDQWLDRPDILADDIKLMKLSHT
NTFSVGIFAWSALEPEEGVYQFEWLDDIFERIHSIGGRVILATPSGARPAWLSQTYPEV
LRVNASRVKQLHGGRHNHCLTSKVYREKTRHINRLLAERYGHHPALLMWHISNEYGGDC
HCESMRPL"
ORIGIN
1 atcctgatat tgtctgcatt tgcgccggag caaaccaaaa acctggtgag acacgccttg
61 aattagtaga aaagaacttg aagattttca aaggcatcgt tagtgaagtc atggcgagcg
121 gatttgacgg cattttctta gtcgcgacgc gaggctggat ggccttcccc attatgattc
181 ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag
241 atgacgacca tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga
301 tcactggacc gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt
361 tggcatggat tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg
421 catggagccg ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac
481 cactccaaga attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc
541 ttggcagaac atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg
601 cagcgttggg tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct
661 aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg
721 aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt
781 ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat
841 ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg
901 ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt
961 accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat
1021 cctctctcgt ttcatcggta tcattacccc catgaacaga aattccccct tacacggagg
1081 catcaagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat cagaagccag
1141 acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc agacatctgt
1201 gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg tttcggtgat
1261 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
1321 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
1381 gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat
1441 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
1501 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
1561 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
1621 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
1681 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
1741 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
1801 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
1861 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
1921 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
1981 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
2041 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
2101 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
2161 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
2221 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
2281 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
2341 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
2401 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
2461 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
2521 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
2581 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
2641 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
2701 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
2761 gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
2821 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
2881 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
2941 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
3001 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
3061 gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag
3121 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
3181 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
3241 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
3301 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
3361 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
3421 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
3481 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaatgatggg
3541 ctaatgtgtg tttacgacaa ttctcacttc atacttttcc atcgtcaggt cgcctgacaa
3601 tatgtctcct gtcattatgt ccttcacact ctgatcaaac gtgaccagct gtttttcttc
3661 cgtgaaattc atgacaaaaa tataatcatt gtcctgatcc tgcctcgctt gtacggagac
3721 gccttttccg tgccgaaccg gaaaaactgg agagagagac aggtctgtga tcagaccctc
3781 atagaaatca cgctgaaatt gatcctccaa acgcgcgccg ataaaatacg ccttgccctg
3841 ctgatactca tggcttgtga ccgctggcgt gcgcgcataa aaatcttctt gatacaccgc
3901 ttccactgaa gctgtcttta catcaatcac ggttgcataa tccttcattt catatatttg
3961 gctgcggtag ctgacagcgt ttcgatcctt cggatacagg gtgtccgttt caagaggctc
4021 aactccaaat atagcttgaa atcgatatct ctgcagtcgc gatgattaat taattcagaa
4081 cgctcggttg ccgccgggcg ttttttatgc agcaatggca agaacgtccc ggggagctcc
4141 taacttatag gggtaacact taaaaaagaa tcaataacga tagaaaccgc tcctaaagca
4201 ggtgcatttt ttcctaacga agaaggcaat agttcacatt tattgtctaa atgagaatgg
4261 actctagaag aaacttcgtt tttaatcgta tttaaaacaa tgggatgaga ttcaattata
4321 tgatttctca agataacagc ttctatatca aatgtattaa ggatattggt taatccaatt
4381 ccgatataaa agccaaagtt ttgaagtgca tttaacattt ctacatcatt tttatttgcg
4441 cgttccacaa tctcttttcg agaaatattc ttttcttctt tagagagcga agccagtaac
4501 gctttttcag aagcatataa ttcccaacag cctcgatttc cacagctgca tttgggtcca
4561 ttaaaatcta tcgtcatatg acccatttcc ccagaaaaac cctgaacacc tttatacaat
4621 tcgttgttaa taacaagtcc agttccaatt ccgatattaa tactgatgta aacgatgttt
4681 tcatagtttt ttgtcatacc aaatactttt tcaccgtatg ctcctgcatt agcttcattt
4741 tcaacaaaaa ccggaacatt aaactcactc tcaattaaaa actgcaaatc tttgatattc
4801 caatttaagt taggcatgaa aataatttgc tgatgacgat ctacaaggcc tggaacacaa
4861 attcctattc cgactagacc ataaggggac tcaggcatat gggttacaaa accatgaata
4921 agtgcaaata aaatctcttt tacttcacta gcggaagaac tagacaagtc agaagtcttc
4981 tcgagaataa tatttccttc taagtcggtt agaattccgt taagatagtc gactcctata
5041 tcaataccaa tcgagtagcc tgcattctta ttaaaaacaa gcattacagg tcttctgccg
5101 cctctagatt gccctgcccc aatttcaaaa ataaaatctt tttcaagcag tgtatttact
5161 tgagaggaga cagtagactt gtttaatcct gtaatctcag agagagttgc cctggagaca
5221 ggggagttct tcaaaatttc atctaatatt aatttttgat tcattttttt tactaaagct
5281 tgatctgcaa tttgaataat aaccactcct ttgtttatcc accgaactaa gttggtgttt
5341 tttgaagctt gaattagata tttaaaagta tcatatctaa tattataact aaattttcta
5401 aaaaaaacat tgaaataaac atttattttg tatatgatga gataaagtta gtttattgga
5461 taaacaaact aactcaatta agatagttga tggataaact tgttcactta aatcaaaggg
5521 ggaaatgaca aatggtccaa actagtgata tctaaaaatc aaagggggaa atgggatcca
5581 tggataagaa atactcaata ggcttagcta tcggcacaaa tagcgtcgga tgggcggtga
5641 tcactgatga atataaggtt ccgtctaaaa agttcaaggt tctgggaaat acagaccgcc
5701 acagtatcaa aaaaaatctt ataggggctc ttttatttga cagtggagag acagcggaag
5761 cgactcgtct caaacggaca gctcgtagaa ggtatacacg tcggaagaat cgtatttgtt
5821 atctacagga gattttttca aatgagatgg cgaaagtaga tgatagtttc tttcatcgac
5881 ttgaagagtc ttttttggtg gaagaagaca agaagcatga acgtcatcct atttttggaa
5941 atatagtaga tgaagttgct tatcatgaga aatatccaac tatctatcat ctgcgaaaaa
6001 aattggtaga ttctactgat aaagcggatt tgcgcttaat ctatttggcc ttagcgcata
6061 tgattaagtt tcgtggtcat tttttgattg agggagattt aaatcctgat aatagtgatg
6121 tggacaaact atttatccag ttggtacaaa cctacaatca attatttgaa gaaaacccta
6181 ttaacgcaag tggagtagat gctaaagcga ttctttctgc acgattgagt aaatcaagac
6241 gattagaaaa tctcattgct cagctccccg gtgagaagaa aaatggctta tttgggaatc
6301 tcattgcttt gtcattgggt ttgaccccta attttaaatc aaattttgat ttggcagaag
6361 atgctaaatt acagctttca aaagatactt acgatgatga tttagataat ttattggcgc
6421 aaattggaga tcaatatgct gatttgtttt tggcagctaa gaatttatca gatgctattt
6481 tactttcaga tatcctaaga gtaaatactg aaataactaa ggctccccta tcagcttcaa
6541 tgattaaacg ctacgatgaa catcatcaag acttgactct tttaaaagct ttagttcgac
6601 aacaacttcc agaaaagtat aaagaaatct tttttgatca atcaaaaaac ggatatgcag
6661 gttatattga tgggggagct agccaagaag aattttataa atttatcaaa ccaattttag
6721 aaaaaatgga tggtactgag gaattattgg tgaaactaaa tcgtgaagat ttgctgcgca
6781 agcaacggac ctttgacaac ggctctattc cccatcaaat tcacttgggt gagctgcatg
6841 ctattttgag aagacaagaa gacttttatc catttttaaa agacaatcgt gagaagattg
6901 aaaaaatctt gacttttcga attccttatt atgttggtcc attggcgcgt ggcaatagtc
6961 gttttgcatg gatgactcgg aagtctgaag aaacaattac cccatggaat tttgaagaag
7021 ttgtcgataa aggtgcttca gctcaatcat ttattgaacg catgacaaac tttgataaaa
7081 atcttccaaa tgaaaaagta ctaccaaaac atagtttgct ttatgagtat tttacggttt
7141 ataacgaatt gacaaaggtc aaatatgtta ctgaaggaat gcgaaaacca gcatttcttt
7201 caggtgaaca gaagaaagcc attgttgatt tactcttcaa aacaaatcga aaagtaaccg
7261 ttaagcaatt aaaagaagat tatttcaaaa aaatagaatg ttttgatagt gttgaaattt
7321 caggagttga agatagattt aatgcttcat taggtaccta ccatgatttg ctaaaaatta
7381 ttaaagataa agattttttg gataatgaag aaaatgaaga tatcttagag gatattgttt
7441 taacattgac cttatttgaa gatagggaga tgattgagga aagacttaaa acatatgctc
7501 acctctttga tgataaggtg atgaaacagc ttaaacgtcg ccgttatact ggttggggac
7561 gtttgtctcg aaaattgatt aatggtatta gggataagca atctggcaaa acaatattag
7621 attttttgaa atcagatggt tttgccaatc gcaattttat gcagctgatc catgatgata
7681 gtttgacatt taaagaagac attcaaaaag cacaagtgtc tggacaaggc gatagtttac
7741 atgaacatat tgcaaattta gctggtagcc ctgctattaa aaaaggtatt ttacagactg
7801 taaaagttgt tgatgaattg gtcaaagtaa tggggcggca taagccagaa aatatcgtta
7861 ttgaaatggc acgtgaaaat cagacaactc aaaagggcca gaaaaattcg cgagagcgta
7921 tgaaacgaat cgaagaaggt atcaaagaat taggaagtca gattcttaaa gagcatcctg
7981 ttgaaaatac tcaattgcaa aatgaaaagc tctatctcta ttatctccaa aatggaagag
8041 acatgtatgt ggaccaagaa ttagatatta atcgtttaag tgattatgat gtcgatgcca
8101 ttgttccaca aagtttcctt aaagacgatt caatagacaa taaggtctta acgcgttctg
8161 ataaaaatcg tggtaaatcg gataacgttc caagtgaaga agtagtcaaa aagatgaaaa
8221 actattggag acaacttcta aacgccaagt taatcactca acgtaagttt gataatttaa
8281 cgaaagctga acgtggaggt ttgagtgaac ttgataaagc tggttttatc aaacgccaat
8341 tggttgaaac tcgccaaatc actaagcatg tggcacaaat tttggatagt cgcatgaata
8401 ctaaatacga tgaaaatgat aaacttattc gagaggttaa agtgattacc ttaaaatcta
8461 aattagtttc tgacttccga aaagatttcc aattctataa agtacgtgag attaacaatt
8521 accatcatgc ccatgatgcg tatctaaatg ccgtcgttgg aactgctttg attaagaaat
8581 atccaaaact tgaatcggag tttgtctatg gtgattataa agtttatgat gttcgtaaaa
8641 tgattgctaa gtctgagcaa gaaataggca aagcaaccgc aaaatatttc ttttactcta
8701 atatcatgaa cttcttcaaa acagaaatta cacttgcaaa tggagagatt cgcaaacgcc
8761 ctctaatcga aactaatggg gaaactggag aaattgtctg ggataaaggg cgagattttg
8821 ccacagtgcg caaagtattg tccatgcccc aagtcaatat tgtcaagaaa acagaagtac
8881 agacaggcgg attctccaag gagtcaattt taccaaaaag aaattcggac aagcttattg
8941 ctcgtaaaaa agactgggat ccaaaaaaat atggtggttt tgatagtcca acggtagctt
9001 attcagtcct agtggttgct aaggtggaaa aagggaaatc gaagaagtta aaatccgtta
9061 aagagttact agggatcaca attatggaaa gaagttcctt tgaaaaaaat ccgattgact
9121 ttttagaagc taaaggatat aaggaagtta aaaaagactt aatcattaaa ctacctaaat
9181 atagtctttt tgagttagaa aacggtcgta aacggatgct ggctagtgcc ggagaattac
9241 aaaaaggaaa tgagctggct ctgccaagca aatatgtgaa ttttttatat ttagctagtc
9301 attatgaaaa gttgaagggt agtccagaag ataacgaaca aaaacaattg tttgtggagc
9361 agcataagca ttatttagat gagattattg agcaaatcag tgaattttct aagcgtgtta
9421 ttttagcaga tgccaattta gataaagttc ttagtgcata taacaaacat agagacaaac
9481 caatacgtga acaagcagaa aatattattc atttatttac gttgacgaat cttggagctc
9541 ccgctgcttt taaatatttt gatacaacaa ttgatcgtaa acgatatacg tctacaaaag
9601 aagttttaga tgccactctt atccatcaat ccatcactgg tctttatgaa acacgcattg
9661 atttgagtca gctaggaggt gactaactcg agtaaggatc tccaggcatc aaataaaacg
9721 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
9781 ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata cctagggata
9841 tattccgctt cctcgctcac tgactcggtc tcggatccga gctcccgcgg gcggccgcac
9901 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
9961 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
10021 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
10081 tcacgaggcc ctttcgtctt caagaattga tcctctagca caaaagaaaa acgaaatgat
10141 acaccaatca gtgcaaaaaa agatataatg ggagataaga cggttcgtgt tcgtgctgac
10201 ttgcaccata tcataaaaat cgaaacagca aagaatggcg gaaacgtaaa agaagttatg
10261 gaaataagac ttagaagcaa acttaagagt gtgttgatag tgcagtatct taaaattttg
10321 tataatagga attgaagtta aattagatgc taaaaatttg taattaagaa ggagtgatta
10381 catgaacaaa aatataaaat attctcaaaa ctttttaacg agtgaaaaag tactcaacca
10441 aataataaaa caattgaatt taaaagaaac cgataccgtt tacgaaattg gaacaggtaa
10501 agggcattta acgacgaaac tggctaaaat aagtaaacag gtaacgtcta ttgaattaga
10561 cagtcatcta ttcaacttat cgtcagaaaa attaaaactg aatactcgtg tcactttaat
10621 tcaccaagat attctacagt ttcaattccc taacaaacag aggtataaaa ttgttgggag
10681 tattccttac catttaagca cacaaattat taaaaaagtg gtttttgaaa gccatgcgtc
10741 tgacatctat ctgattgttg aagaaggatt ctacaagcgt accttggata ttcaccgaac
10801 actagggttg ctcttgcaca ctcaagtctc gattcagcaa ttgcttaagc tgccagcgga
10861 atgctttcat cctaaaccaa aagtaaacag tgtcttaata aaacttaccc gccataccac
10921 agatgttcca gataaatatt ggaagctata tacgtacttt gtttcaaaat gggtcaatcg
10981 agaatatcgt caactgttta ctaaaaatca gtttcatcaa gcaatgaaac acgccaaagt
11041 aaacaattta agtaccgtta cttatgagca agtattgtct atttttaata gttatctatt
11101 atttaacggg aggaaataat tctatgagtc gcttttgtaa atttggaaag ttacacgtta
11161 ctaaagggaa tgtagataaa ttattaggta tactactgac agcttccaag gagctaaaga
11221 ggtccctaga ctctagaccc ggggatctct gcagtgagat ctggtaatga ctctctagct
11281 tgaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
11341 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gctctagcta agcagaaggc
11401 catcctgacg gatggccttt ttgcgtttct acaaactctt gttaactcta gagctgcctg
11461 ccgcgtttcg gtgatgaaga tcttcccgat gattaattaa ttcagaacgc tcggttgccg
11521 ccgggcgttt tttatgcagc aatggcaaga acgttgctct agagcggccg catcgattca
11581 cagtggcaat ctcccccgta ttcgtttgaa atgtgccaca ttaacagcgc cgggtgatgt
11641 ccgtatcgtt ctgctaataa gcggttgatg tgccgtgttt tttctcggta gactttagat
11701 gtgaggcagt ggttgtgcct tccgccgtgc agctgtttga cgcgggaggc attgacgcgc
11761 aaaacttccg gataggtttg cgacagccag gccggacggg ctccgctcgg cgttgctaat
11821 atgacccggc cgcctatact gtgaatccgc tcaaaaatat catccagcca ttcaaattga
11881 tatacgccct cctccggctc aagtgcgctc catgcaaaaa tgccgacaga aaacgtattc
11941 gtatgagaaa gcttcatcag tttgatatcg tcagctaaaa tatcgggccg atccagccac
12001 tgatcggggt tgtagtctcc cccatggagc ataaattttg cttttgttac gtgcgttttt
12061 tcaagctttg acatcacccc
//
质粒菌株产品操作说明书
一、扩增流程
收到产品后，请先根据产品管壁标签来判断产品形式，并在扩增前准确查找该质粒菌株的抗性、感受态和培养温度。
1、质粒干粉（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
①收到质粒干粉后请先5000rpm离心1min，再加入20μl ddH2O去离子水溶解质粒；
②取1支100μl 感受态于冰上解冻10min，加入2μl质粒，再冰浴30min后，42℃热激60s，不要搅动，再冰浴2min；（从第二步开始均要在超净工作台中无菌操作）
③加入900μl无抗的LB液体培养基，180rpm震荡37℃培养45min；
④6000rpm离心5min，仅留100μl上清液重悬细菌沉淀，并涂布至目标质粒抗性的LB平板上；（可使用本平台的平板涂布专用玻璃珠进行涂布，可以比传统涂布方法获得更多转化子）
⑤将平板正向培养1h，再倒置37℃培养14h。如果要求是30度则培养20h；
（菌落过多则将质粒稀释后再转化。没有菌落则加入10μl质粒转化。另不要直接转表达感受态，要先转克隆感受态，重提质粒后再导入表达感受态）
⑥挑取单菌落至LB液体培养基中，加入对应抗生素，220rpm震荡培养14h，根据实验需要和质粒提取试剂盒说明书提取质粒。
2、甘油菌种（冰袋运输，存于-80℃，保质期90天，请务必划线挑单克隆培养）
四区划线后挑单菌落培养，酵母菌需要先液体复苏再四区划线，再挑单菌落液体培养。
3、穿刺菌种（冰袋运输，存于4℃，保质期7天）
穿刺接种，液体培养后四区划线，再挑单菌落液体培养。
4、菌落平板（冰袋运输，存于4℃，保质期7天）
直接挑取单菌落至液体培养基中。
5、液体质粒（冰袋运输，存于-20℃，保质期90天）
单独提取的液体质粒收到后可直接使用。
6、滤纸质粒（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
收到货后将滤纸画圈部分剪下放入EP管中，加100ul无菌水将滤纸浸湿并浸泡5min，吸取5ul质粒转化，离心全涂。
二、转化图片
名称：pJMP1枯草编辑质粒
别称: Plasmid#79873;pAX01-dCas9
| 启动子: | xylA |
|---|---|
| 原核抗性: | Amp |
| 筛选标记: | Ery |
| 克隆菌株: | DH5a |
| 培养条件: | 37度 |
| 表达宿主: | 枯草杆菌 |
质粒属性
| 载体宿主： | 枯草杆菌 |
|---|---|
| 载体用途： | 基因敲除 |
| 基因种属： | |
| 基因类型： | |
| 原核抗性： | Amp |
| 筛选标记： | |
| 荧光蛋白： |
pJMP1质粒是一个枯草杆菌基因编辑质粒。
质粒图谱
质粒序列
LOCUS Exported 12080 bp ds-DNA circular SYN 14-MAY-2018
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..12080
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 1087..1278
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 1380..1520
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(1706..2294)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2465..3325)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3326..3430)
/gene="bla"
/note="AmpR promoter"
CDS complement(3541..3948)
/codon_start=1
/note="'lacA"
/translation="MKDYATVIDVKTASVEAVYQEDFYARTPAVTSHEYQQGKAYFIGA
RLEDQFQRDFYEGLITDLSLSPVFPVRHGKGVSVQARQDQDNDYIFVMNFTEEKQLVTF
DQSVKDIMTGDILSGDLTMEKYEVRIVVNTH"
terminator 4074..4108
/note="lambda t0 terminator"
/note="minimal transcription terminator from phage lambda
(Scholtissek and Grosse, 1987)"
CDS complement(4140..5264)
/codon_start=1
/note="bme xylR"
/translation="MNQKLILDEILKNSPVSRATLSEITGLNKSTVSSQVNTLLEKDFI
FEIGAGQSRGGRRPVMLVFNKNAGYSIGIDIGVDYLNGILTDLEGNIILEKTSDLSSSS
ASEVKEILFALIHGFVTHMPESPYGLVGIGICVPGLVDRHQQIIFMPNLNWNIKDLQFL
IESEFNVPVFVENEANAGAYGEKVFGMTKNYENIVYISINIGIGTGLVINNELYKGVQG
FSGEMGHMTIDFNGPKCSCGNRGCWELYASEKALLASLSKEEKNISRKEIVERANKNDV
EMLNALQNFGFYIGIGLTNILNTFDIEAVILRNHIIESHPIVLNTIKNEVSSRVHSHLD
NKCELLPSSLGKNAPALGAVSIVIDSFLSVTPIS"
CDS 5580..9686
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/note="dCas9"
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the D10A mutation in the RuvC
catalytic domain and the H840A mutation in the HNH
catalytic domain"
/translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNL-SDAILLS-DILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLS-DYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
terminator 9710..9781
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 9797..9824
/note="T7Te terminator"
/note="phage T7 early transcription terminator"
promoter complement(9899..10003)
/gene="bla"
/note="AmpR promoter"
CDS 10382..11119
/codon_start=1
/note="Erm"
/translation="MNKNIKYSQNFLTSEKVLNQIIKQLNLKETDTVYEIGTGKGHLTT
KLAKISKQVTSIELDSHLFNLSSEKLKLNTRVTLIHQDILQFQFPNKQRYKIVGSIPYH
LSTQIIKKVVFESHASDIYLIVEEGFYKRTLDIHRTLGLLLHTQVSIQQLLKLPAECFH
PKPKVNSVLIKLTRHTTDVPDKYWKLYTYFVSKWVNREYRQLFTKNQFHQAMKHAKVNN
LSTVTYEQVLSIFNSYLLFNGRK"
terminator 11289..11375
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 11394..11421
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 11501..11535
/note="lambda t0 terminator"
/note="minimal transcription terminator from phage lambda
(Scholtissek and Grosse, 1987)"
CDS complement(11559..12074)
/codon_start=1
/note="lacA'"
/translation="MSKLEKTHVTKAKFMLHGGDYNPDQWLDRPDILADDIKLMKLSHT
NTFSVGIFAWSALEPEEGVYQFEWLDDIFERIHSIGGRVILATPSGARPAWLSQTYPEV
LRVNASRVKQLHGGRHNHCLTSKVYREKTRHINRLLAERYGHHPALLMWHISNEYGGDC
HCESMRPL"
ORIGIN
1 atcctgatat tgtctgcatt tgcgccggag caaaccaaaa acctggtgag acacgccttg
61 aattagtaga aaagaacttg aagattttca aaggcatcgt tagtgaagtc atggcgagcg
121 gatttgacgg cattttctta gtcgcgacgc gaggctggat ggccttcccc attatgattc
181 ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag
241 atgacgacca tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga
301 tcactggacc gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt
361 tggcatggat tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg
421 catggagccg ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac
481 cactccaaga attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc
541 ttggcagaac atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg
601 cagcgttggg tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct
661 aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg
721 aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt
781 ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat
841 ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg
901 ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt
961 accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat
1021 cctctctcgt ttcatcggta tcattacccc catgaacaga aattccccct tacacggagg
1081 catcaagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat cagaagccag
1141 acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc agacatctgt
1201 gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg tttcggtgat
1261 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
1321 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
1381 gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat
1441 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
1501 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
1561 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
1621 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
1681 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
1741 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
1801 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
1861 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
1921 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
1981 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
2041 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
2101 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
2161 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
2221 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
2281 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
2341 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
2401 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
2461 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
2521 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
2581 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
2641 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
2701 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
2761 gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
2821 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
2881 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
2941 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
3001 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
3061 gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag
3121 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
3181 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
3241 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
3301 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
3361 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
3421 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
3481 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaatgatggg
3541 ctaatgtgtg tttacgacaa ttctcacttc atacttttcc atcgtcaggt cgcctgacaa
3601 tatgtctcct gtcattatgt ccttcacact ctgatcaaac gtgaccagct gtttttcttc
3661 cgtgaaattc atgacaaaaa tataatcatt gtcctgatcc tgcctcgctt gtacggagac
3721 gccttttccg tgccgaaccg gaaaaactgg agagagagac aggtctgtga tcagaccctc
3781 atagaaatca cgctgaaatt gatcctccaa acgcgcgccg ataaaatacg ccttgccctg
3841 ctgatactca tggcttgtga ccgctggcgt gcgcgcataa aaatcttctt gatacaccgc
3901 ttccactgaa gctgtcttta catcaatcac ggttgcataa tccttcattt catatatttg
3961 gctgcggtag ctgacagcgt ttcgatcctt cggatacagg gtgtccgttt caagaggctc
4021 aactccaaat atagcttgaa atcgatatct ctgcagtcgc gatgattaat taattcagaa
4081 cgctcggttg ccgccgggcg ttttttatgc agcaatggca agaacgtccc ggggagctcc
4141 taacttatag gggtaacact taaaaaagaa tcaataacga tagaaaccgc tcctaaagca
4201 ggtgcatttt ttcctaacga agaaggcaat agttcacatt tattgtctaa atgagaatgg
4261 actctagaag aaacttcgtt tttaatcgta tttaaaacaa tgggatgaga ttcaattata
4321 tgatttctca agataacagc ttctatatca aatgtattaa ggatattggt taatccaatt
4381 ccgatataaa agccaaagtt ttgaagtgca tttaacattt ctacatcatt tttatttgcg
4441 cgttccacaa tctcttttcg agaaatattc ttttcttctt tagagagcga agccagtaac
4501 gctttttcag aagcatataa ttcccaacag cctcgatttc cacagctgca tttgggtcca
4561 ttaaaatcta tcgtcatatg acccatttcc ccagaaaaac cctgaacacc tttatacaat
4621 tcgttgttaa taacaagtcc agttccaatt ccgatattaa tactgatgta aacgatgttt
4681 tcatagtttt ttgtcatacc aaatactttt tcaccgtatg ctcctgcatt agcttcattt
4741 tcaacaaaaa ccggaacatt aaactcactc tcaattaaaa actgcaaatc tttgatattc
4801 caatttaagt taggcatgaa aataatttgc tgatgacgat ctacaaggcc tggaacacaa
4861 attcctattc cgactagacc ataaggggac tcaggcatat gggttacaaa accatgaata
4921 agtgcaaata aaatctcttt tacttcacta gcggaagaac tagacaagtc agaagtcttc
4981 tcgagaataa tatttccttc taagtcggtt agaattccgt taagatagtc gactcctata
5041 tcaataccaa tcgagtagcc tgcattctta ttaaaaacaa gcattacagg tcttctgccg
5101 cctctagatt gccctgcccc aatttcaaaa ataaaatctt tttcaagcag tgtatttact
5161 tgagaggaga cagtagactt gtttaatcct gtaatctcag agagagttgc cctggagaca
5221 ggggagttct tcaaaatttc atctaatatt aatttttgat tcattttttt tactaaagct
5281 tgatctgcaa tttgaataat aaccactcct ttgtttatcc accgaactaa gttggtgttt
5341 tttgaagctt gaattagata tttaaaagta tcatatctaa tattataact aaattttcta
5401 aaaaaaacat tgaaataaac atttattttg tatatgatga gataaagtta gtttattgga
5461 taaacaaact aactcaatta agatagttga tggataaact tgttcactta aatcaaaggg
5521 ggaaatgaca aatggtccaa actagtgata tctaaaaatc aaagggggaa atgggatcca
5581 tggataagaa atactcaata ggcttagcta tcggcacaaa tagcgtcgga tgggcggtga
5641 tcactgatga atataaggtt ccgtctaaaa agttcaaggt tctgggaaat acagaccgcc
5701 acagtatcaa aaaaaatctt ataggggctc ttttatttga cagtggagag acagcggaag
5761 cgactcgtct caaacggaca gctcgtagaa ggtatacacg tcggaagaat cgtatttgtt
5821 atctacagga gattttttca aatgagatgg cgaaagtaga tgatagtttc tttcatcgac
5881 ttgaagagtc ttttttggtg gaagaagaca agaagcatga acgtcatcct atttttggaa
5941 atatagtaga tgaagttgct tatcatgaga aatatccaac tatctatcat ctgcgaaaaa
6001 aattggtaga ttctactgat aaagcggatt tgcgcttaat ctatttggcc ttagcgcata
6061 tgattaagtt tcgtggtcat tttttgattg agggagattt aaatcctgat aatagtgatg
6121 tggacaaact atttatccag ttggtacaaa cctacaatca attatttgaa gaaaacccta
6181 ttaacgcaag tggagtagat gctaaagcga ttctttctgc acgattgagt aaatcaagac
6241 gattagaaaa tctcattgct cagctccccg gtgagaagaa aaatggctta tttgggaatc
6301 tcattgcttt gtcattgggt ttgaccccta attttaaatc aaattttgat ttggcagaag
6361 atgctaaatt acagctttca aaagatactt acgatgatga tttagataat ttattggcgc
6421 aaattggaga tcaatatgct gatttgtttt tggcagctaa gaatttatca gatgctattt
6481 tactttcaga tatcctaaga gtaaatactg aaataactaa ggctccccta tcagcttcaa
6541 tgattaaacg ctacgatgaa catcatcaag acttgactct tttaaaagct ttagttcgac
6601 aacaacttcc agaaaagtat aaagaaatct tttttgatca atcaaaaaac ggatatgcag
6661 gttatattga tgggggagct agccaagaag aattttataa atttatcaaa ccaattttag
6721 aaaaaatgga tggtactgag gaattattgg tgaaactaaa tcgtgaagat ttgctgcgca
6781 agcaacggac ctttgacaac ggctctattc cccatcaaat tcacttgggt gagctgcatg
6841 ctattttgag aagacaagaa gacttttatc catttttaaa agacaatcgt gagaagattg
6901 aaaaaatctt gacttttcga attccttatt atgttggtcc attggcgcgt ggcaatagtc
6961 gttttgcatg gatgactcgg aagtctgaag aaacaattac cccatggaat tttgaagaag
7021 ttgtcgataa aggtgcttca gctcaatcat ttattgaacg catgacaaac tttgataaaa
7081 atcttccaaa tgaaaaagta ctaccaaaac atagtttgct ttatgagtat tttacggttt
7141 ataacgaatt gacaaaggtc aaatatgtta ctgaaggaat gcgaaaacca gcatttcttt
7201 caggtgaaca gaagaaagcc attgttgatt tactcttcaa aacaaatcga aaagtaaccg
7261 ttaagcaatt aaaagaagat tatttcaaaa aaatagaatg ttttgatagt gttgaaattt
7321 caggagttga agatagattt aatgcttcat taggtaccta ccatgatttg ctaaaaatta
7381 ttaaagataa agattttttg gataatgaag aaaatgaaga tatcttagag gatattgttt
7441 taacattgac cttatttgaa gatagggaga tgattgagga aagacttaaa acatatgctc
7501 acctctttga tgataaggtg atgaaacagc ttaaacgtcg ccgttatact ggttggggac
7561 gtttgtctcg aaaattgatt aatggtatta gggataagca atctggcaaa acaatattag
7621 attttttgaa atcagatggt tttgccaatc gcaattttat gcagctgatc catgatgata
7681 gtttgacatt taaagaagac attcaaaaag cacaagtgtc tggacaaggc gatagtttac
7741 atgaacatat tgcaaattta gctggtagcc ctgctattaa aaaaggtatt ttacagactg
7801 taaaagttgt tgatgaattg gtcaaagtaa tggggcggca taagccagaa aatatcgtta
7861 ttgaaatggc acgtgaaaat cagacaactc aaaagggcca gaaaaattcg cgagagcgta
7921 tgaaacgaat cgaagaaggt atcaaagaat taggaagtca gattcttaaa gagcatcctg
7981 ttgaaaatac tcaattgcaa aatgaaaagc tctatctcta ttatctccaa aatggaagag
8041 acatgtatgt ggaccaagaa ttagatatta atcgtttaag tgattatgat gtcgatgcca
8101 ttgttccaca aagtttcctt aaagacgatt caatagacaa taaggtctta acgcgttctg
8161 ataaaaatcg tggtaaatcg gataacgttc caagtgaaga agtagtcaaa aagatgaaaa
8221 actattggag acaacttcta aacgccaagt taatcactca acgtaagttt gataatttaa
8281 cgaaagctga acgtggaggt ttgagtgaac ttgataaagc tggttttatc aaacgccaat
8341 tggttgaaac tcgccaaatc actaagcatg tggcacaaat tttggatagt cgcatgaata
8401 ctaaatacga tgaaaatgat aaacttattc gagaggttaa agtgattacc ttaaaatcta
8461 aattagtttc tgacttccga aaagatttcc aattctataa agtacgtgag attaacaatt
8521 accatcatgc ccatgatgcg tatctaaatg ccgtcgttgg aactgctttg attaagaaat
8581 atccaaaact tgaatcggag tttgtctatg gtgattataa agtttatgat gttcgtaaaa
8641 tgattgctaa gtctgagcaa gaaataggca aagcaaccgc aaaatatttc ttttactcta
8701 atatcatgaa cttcttcaaa acagaaatta cacttgcaaa tggagagatt cgcaaacgcc
8761 ctctaatcga aactaatggg gaaactggag aaattgtctg ggataaaggg cgagattttg
8821 ccacagtgcg caaagtattg tccatgcccc aagtcaatat tgtcaagaaa acagaagtac
8881 agacaggcgg attctccaag gagtcaattt taccaaaaag aaattcggac aagcttattg
8941 ctcgtaaaaa agactgggat ccaaaaaaat atggtggttt tgatagtcca acggtagctt
9001 attcagtcct agtggttgct aaggtggaaa aagggaaatc gaagaagtta aaatccgtta
9061 aagagttact agggatcaca attatggaaa gaagttcctt tgaaaaaaat ccgattgact
9121 ttttagaagc taaaggatat aaggaagtta aaaaagactt aatcattaaa ctacctaaat
9181 atagtctttt tgagttagaa aacggtcgta aacggatgct ggctagtgcc ggagaattac
9241 aaaaaggaaa tgagctggct ctgccaagca aatatgtgaa ttttttatat ttagctagtc
9301 attatgaaaa gttgaagggt agtccagaag ataacgaaca aaaacaattg tttgtggagc
9361 agcataagca ttatttagat gagattattg agcaaatcag tgaattttct aagcgtgtta
9421 ttttagcaga tgccaattta gataaagttc ttagtgcata taacaaacat agagacaaac
9481 caatacgtga acaagcagaa aatattattc atttatttac gttgacgaat cttggagctc
9541 ccgctgcttt taaatatttt gatacaacaa ttgatcgtaa acgatatacg tctacaaaag
9601 aagttttaga tgccactctt atccatcaat ccatcactgg tctttatgaa acacgcattg
9661 atttgagtca gctaggaggt gactaactcg agtaaggatc tccaggcatc aaataaaacg
9721 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
9781 ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata cctagggata
9841 tattccgctt cctcgctcac tgactcggtc tcggatccga gctcccgcgg gcggccgcac
9901 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
9961 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
10021 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
10081 tcacgaggcc ctttcgtctt caagaattga tcctctagca caaaagaaaa acgaaatgat
10141 acaccaatca gtgcaaaaaa agatataatg ggagataaga cggttcgtgt tcgtgctgac
10201 ttgcaccata tcataaaaat cgaaacagca aagaatggcg gaaacgtaaa agaagttatg
10261 gaaataagac ttagaagcaa acttaagagt gtgttgatag tgcagtatct taaaattttg
10321 tataatagga attgaagtta aattagatgc taaaaatttg taattaagaa ggagtgatta
10381 catgaacaaa aatataaaat attctcaaaa ctttttaacg agtgaaaaag tactcaacca
10441 aataataaaa caattgaatt taaaagaaac cgataccgtt tacgaaattg gaacaggtaa
10501 agggcattta acgacgaaac tggctaaaat aagtaaacag gtaacgtcta ttgaattaga
10561 cagtcatcta ttcaacttat cgtcagaaaa attaaaactg aatactcgtg tcactttaat
10621 tcaccaagat attctacagt ttcaattccc taacaaacag aggtataaaa ttgttgggag
10681 tattccttac catttaagca cacaaattat taaaaaagtg gtttttgaaa gccatgcgtc
10741 tgacatctat ctgattgttg aagaaggatt ctacaagcgt accttggata ttcaccgaac
10801 actagggttg ctcttgcaca ctcaagtctc gattcagcaa ttgcttaagc tgccagcgga
10861 atgctttcat cctaaaccaa aagtaaacag tgtcttaata aaacttaccc gccataccac
10921 agatgttcca gataaatatt ggaagctata tacgtacttt gtttcaaaat gggtcaatcg
10981 agaatatcgt caactgttta ctaaaaatca gtttcatcaa gcaatgaaac acgccaaagt
11041 aaacaattta agtaccgtta cttatgagca agtattgtct atttttaata gttatctatt
11101 atttaacggg aggaaataat tctatgagtc gcttttgtaa atttggaaag ttacacgtta
11161 ctaaagggaa tgtagataaa ttattaggta tactactgac agcttccaag gagctaaaga
11221 ggtccctaga ctctagaccc ggggatctct gcagtgagat ctggtaatga ctctctagct
11281 tgaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
11341 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gctctagcta agcagaaggc
11401 catcctgacg gatggccttt ttgcgtttct acaaactctt gttaactcta gagctgcctg
11461 ccgcgtttcg gtgatgaaga tcttcccgat gattaattaa ttcagaacgc tcggttgccg
11521 ccgggcgttt tttatgcagc aatggcaaga acgttgctct agagcggccg catcgattca
11581 cagtggcaat ctcccccgta ttcgtttgaa atgtgccaca ttaacagcgc cgggtgatgt
11641 ccgtatcgtt ctgctaataa gcggttgatg tgccgtgttt tttctcggta gactttagat
11701 gtgaggcagt ggttgtgcct tccgccgtgc agctgtttga cgcgggaggc attgacgcgc
11761 aaaacttccg gataggtttg cgacagccag gccggacggg ctccgctcgg cgttgctaat
11821 atgacccggc cgcctatact gtgaatccgc tcaaaaatat catccagcca ttcaaattga
11881 tatacgccct cctccggctc aagtgcgctc catgcaaaaa tgccgacaga aaacgtattc
11941 gtatgagaaa gcttcatcag tttgatatcg tcagctaaaa tatcgggccg atccagccac
12001 tgatcggggt tgtagtctcc cccatggagc ataaattttg cttttgttac gtgcgttttt
12061 tcaagctttg acatcacccc
//
质粒菌株产品操作说明书
一、扩增流程
收到产品后，请先根据产品管壁标签来判断产品形式，并在扩增前准确查找该质粒菌株的抗性、感受态和培养温度。
1、质粒干粉（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
①收到质粒干粉后请先5000rpm离心1min，再加入20μl ddH2O去离子水溶解质粒；
②取1支100μl 感受态于冰上解冻10min，加入2μl质粒，再冰浴30min后，42℃热激60s，不要搅动，再冰浴2min；（从第二步开始均要在超净工作台中无菌操作）
③加入900μl无抗的LB液体培养基，180rpm震荡37℃培养45min；
④6000rpm离心5min，仅留100μl上清液重悬细菌沉淀，并涂布至目标质粒抗性的LB平板上；（可使用本平台的平板涂布专用玻璃珠进行涂布，可以比传统涂布方法获得更多转化子）
⑤将平板正向培养1h，再倒置37℃培养14h。如果要求是30度则培养20h；
（菌落过多则将质粒稀释后再转化。没有菌落则加入10μl质粒转化。另不要直接转表达感受态，要先转克隆感受态，重提质粒后再导入表达感受态）
⑥挑取单菌落至LB液体培养基中，加入对应抗生素，220rpm震荡培养14h，根据实验需要和质粒提取试剂盒说明书提取质粒。
2、甘油菌种（冰袋运输，存于-80℃，保质期90天，请务必划线挑单克隆培养）
四区划线后挑单菌落培养，酵母菌需要先液体复苏再四区划线，再挑单菌落液体培养。
3、穿刺菌种（冰袋运输，存于4℃，保质期7天）
穿刺接种，液体培养后四区划线，再挑单菌落液体培养。
4、菌落平板（冰袋运输，存于4℃，保质期7天）
直接挑取单菌落至液体培养基中。
5、液体质粒（冰袋运输，存于-20℃，保质期90天）
单独提取的液体质粒收到后可直接使用。
6、滤纸质粒（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
收到货后将滤纸画圈部分剪下放入EP管中，加100ul无菌水将滤纸浸湿并浸泡5min，吸取5ul质粒转化，离心全涂。
二、转化图片
| P3352/pCMV-SPORT6-PGAM5(1同义突变1点突变）人源基因质粒 |
| P3353/pCMV-SPORT6-POMP人源基因质粒 |
| P3354/pCMV-SPORT6-G2E3人源基因质粒 |
| P3355/pCMV-SPORT6-PUS3（2点突变）人源基因质粒 |
| P3356/pCMV-SPORT6-RTCB（1同义突变）人源基因质粒 |
| P3357/pCMV-SPORT6-HNRNPC人源基因质粒 |
| P3358/pCMV-SPORT6-ATP5F1C人源基因质粒 |
| P3359/pCMV-SPORT6-APH1A-C706A人源基因质粒 |
| P3360/pCMV-SPORT6-FAM50B人源基因质粒 |
| P3361/pCMV-SPORT6-ATG13人源基因质粒 |
风险提示：丁香通仅作为第三方平台，为商家信息发布提供平台空间。用户咨询产品时请注意保护个人信息及财产安全，合理判断，谨慎选购商品，商家和用户对交易行为负责。对于医疗器械类产品，请先查证核实企业经营资质和医疗器械产品注册证情况。
文献和实验相关实验
Marburg遗传背景清楚，是研究枯草芽孢杆菌的标准菌株；AS1作为胞内表达的备用菌株，常用于表达外源蛋白。而胞外蛋白酶缺陷菌株WB800N能有效解决由于枯草芽孢杆菌丰富的蛋白酶导致的蛋白表达产物降解问题。 2 给力的表达载体 大部分的枯草芽孢杆菌不含内源性质粒，并且一般不能识别大肠杆菌中的启动子。最初的枯草芽孢杆菌载体源于金黄色葡萄球菌，其结构不稳定并易丢失。现在普遍使用的质粒载体是由枯草芽孢杆菌隐性质粒与大肠杆菌质粒构成的穿梭载体，可在大肠杆菌中完成基因片段的连接克隆
1、质粒的概念 质粒（Plasmid）是一种存在于细菌、酵母等微生物细胞中的小型环状双链 DNA 分子，能够独立于宿主染色体进行自我复制，并通常携带对宿主生存非必需但具有特定功能的基因（如抗生素抗性基因）。质粒是分子生物学和基因工程中最重要的工具之一。 质粒根据其用途可以分为多种不同的类型，如用于保存序列信息的克隆质粒（如：pUC 系列质粒），用于表达蛋白的质粒（如哺乳动物细胞表达所用的 pcDNA3.1 系列质粒），用于基因编辑的质粒（如哺乳动物细胞基因编辑所用的 espCas9-2A
基因编辑再次升级！领域大牛刘如谦 Cell 发文开发新工具，可安全高效进行体内基因编辑
选获得 v3.4 BE-eVLPs 系统。 最后，研究人员发现，不同包装质粒间的化学计量比也会影响 BE-eVLP 系统的编辑效果。为了调节这种化学计量学，他们改变了 gag-3xNES-ABE8e 与转染 VLP 的野生型 MMLV gag-pro-pol 质粒的比例。结果发现 gag-pro-pol 质粒与 gag-3XNES-ABE8e 质粒的摩尔比例为 1:3 时获得最佳的碱基编辑效率，并命名为第四代 BE-eVLPs 系统（v4 BE-eVLPs）。 随后的实验也证明，v
pJMP1枯草编辑质粒
￥100 - 1000
