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- 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：
NM_000132.3;AHF;F8B;F8C;HEMA;FVIII;D
- 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：pCDNA4-F8人源基因质粒
别称: NM_000132.3;AHF;F8B;F8C;HEMA;FVIII;D
启动子：CMV
复制子: pUC
原核抗性: Amp
筛选标记：Zeo
克隆菌株: DH5a
培养条件: 37度
质粒属性
载体宿主：哺乳细胞
载体用途：蛋白表达
基因种属：人
基因类型：ORF
原核抗性：Amp
真核抗性：Zeo
荧光蛋白：
质粒简介
pCDNA3.0-HA-p53质粒由上海烜雅生物科技有限公司提供
质粒图谱
质粒序列
LOCUS Exported 12086 bp ds-DNA circular SYN 21-JAN-2018
FEATURES Location/Qualifiers
source 1..12086
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 928..7983
/codon_start=1
/label=F8
/translation="MQIELSTCFFLCLLRFCFSATRRYYLGAVELSWDYMQSDLGELPV
DARFPPRVPKSFPFNTSVVYKKTLFVEFTDHLFNIAKPRPPWMGLLGPTIQAEVYDTVV
ITLKNMASHPVSLHAVGVSYWKASEGAEYDDQTSQREKEDDKVFPGGSHTYVWQVLKEN
GPM-ASDPLCLTYSYLSHVDLVKDLNSGLIGALLVCREGSLAKEKTQTLHKFILLFAVFD
EGKSWHSETKNSLMQDRDAASARAWPKMHTVNGYVNRSLPGLIGCHRKSVYWHVIGMGT
TPEVHSIFLEGHTFLVRNHRQASLEISPITFLTAQTLLMDLGQFLLFCHISSHQHDGME
AYVKVDSCPEEPQLRMKNNEEAEDYDDDLTDSEMDVVRFDDDNSPSFIQIRSVAKKHPK
TWVHYIAAEEEDWDYAPLVLAPDDRSYKSQYLNNGPQRIGRKYKKVRFMAYTDETFKTR
EAIQHESGILGPLLYGEVGDTLLIIFKNQASRPYNIYPHGITDVRPLYSRRLPKGVKHL
KDFPILPGEIFKYKWTVTVEDGPTKSDPRCLTRYYSSFVNMERDLASGLIGPLLICYKE
SVDQRGNQIMSDKRNVILFSVFDENRSWYLTENIQRFLPNPAGVQLEDPEFQASNIMHS
INGYVFDSLQLSVCLHEVAYWYILSIGAQTDFLSVFFSGYTFKHKMVYEDTLTLFPFSG
ETVFMSMENPGLWILGCHNSDFRNRGMTALLKVSSCDKNTGDYYEDSYEDISAYLLSKN
NAIEPRSFSQNSRHPSTRQKQFNATTIPENDIEKTDPWFAHRTPMPKIQNVSSSDLLML
LRQSPTPHGLSL-SDLQEAKYETFSDDPSPGAIDSNNSLSEMTHFRPQLHHSGDMVFTPE
SGLQLRLNEKLGTTAATELKKLDFKVSSTSNNLISTIPSDNLAAGTDNTSSLGPPSMPV
HYDSQLDTTLFGKKSSPLTESGGPLSLSEENNDSKLLESGLMNSQESSWGKNVSSTESG
RLFKGKRAHGPALLTKDNALFKVSISLLKTNKTSNNSATNRKTHIDGPSLLIENSPSVW
QNILESDTEFKKVTPLIHDRMLMDKNATALRLNHMSNKTTSSKNMEMVQQKKEGPIPPD
AQNPDMSFFKMLFLPESARWIQRTHGKNSLNSGQGPSPKQLVSLGPEKSVEGQNFLSEK
NKVVVGKGEFTKDVGLKEMVFPSSRNLFLTNLDNLHENNTHNQEKKIQEEIEKKETLIQ
ENVVLPQIHTVTGTKNFMKNLFLLSTRQNVEGSYDGAYAPVLQDFRSLNDSTNRTKKHT
AHFSKKGEEENLEGLGNQTKQIVEKYACTTRISPNTSQQNFVTQRSKRALKQFRLPLEE
TELEKRIIVDDTSTQWSKNMKHLTPSTLTQIDYNEKEKGAITQSPL-SDCLTRSHSIPQA
NRSPLPIAKVSSFPSIRPIYLTRVLFQDNSSHLPAASYRKKDSGVQESSHFLQGAKKNN
LSLAILTLEMTGDQREVGSLGTSATNSVTYKKVENTVLPKPDLPKTSGKVELLPKVHIY
QKDLFPTETSNGSPGHLDLVEGSLLQGTEGAIKWNEANRPGKVPFLRVATESSAKTPSK
LLDPLAWDNHYGTQIPKEEWKSQEKSPEKTAFKKKDTILSLNACESNHAIAAINEGQNK
PEIEVTWAKQGRTERLCSQNPPVLKRHQREITRTTLQSDQEEIDYDDTISVEMKKEDFD
IYDEDENQSPRSFQKKTRHYFIAAVERLWDYGMSSSPHVLRNRAQSGSVPQFKKVVFQE
FTDGSFTQPLYRGELNEHLGLLGPYIRAEVEDNIMVTFRNQASRPYSFYSSLISYEEDQ
RQGAEPRKNFVKPNETKTYFWKVQHHMAPTKDEFDCKAWAYFSDVDLEKDVHSGLIGPL
LVCHTNTLNPAHGRQVTVQEFALFFTIFDETKSWYFTENMERNCRAPCNIQMEDPTFKE
NYRFHAINGYIMDTLPGLVMAQDQRIRWYLLSMGSNENIHSIHFSGHVFTVRKKEEYKM
ALYNLYPGVFETVEMLPSKAGIWRVECLIGEHLHAGMSTLFLVYSNKCQTPLGMASGHI
RDFQITASGQYGQWAPKLARLHYSGSINAWSTKEPFSWIKVDLLAPMIIHGIKTQGARQ
KFSSLYISQFIIMYSLDGKKWQTYRGNSTGTLMVFFGNVDSSGIKHNIFNPPIIARYIR
LHPTHYSIRSTLRMELMGCDLNSCSMPLGMESKAISDAQITASSYFTNMFATWSPSKAR
LHLQGRSNAWRPQVNNPKEWLQVDFQKTMKVTGVTTQGVKSLLTSMYVKEFLISSSQDG
HQWTLFFQNGKVKVFQGNQDSFTPVVNSLDPPLLTRYLRIHPQSWVHQIALRMEVLGCE
AQDLY"
polyA_signal 8099..8323
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 8369..8797
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 8811..9141
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 9189..9236
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 9255..9629
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=Zeo
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 9759..9880
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(10331..10919)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(11090..11950)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(11951..12055)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggaacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
901 taagcttcgc gacgtacggc cgccaccatg caaatagagc tctccacctg cttctttctg
961 tgccttttgc gattctgctt tagtgccacc agaagatact acctgggtgc agtggaactg
1021 tcatgggact atatgcaaag tgatctcggt gagctgcctg tggacgcaag atttcctcct
1081 agagtgccaa aatcttttcc attcaacacc tcagtcgtgt acaaaaagac tctgtttgta
1141 gaattcacgg atcacctttt caacatcgct aagccaaggc caccctggat gggtctgcta
1201 ggtcctacca tccaggctga ggtttatgat acagtggtca ttacacttaa gaacatggct
1261 tcccatcctg tcagtcttca tgctgttggt gtatcctact ggaaagcttc tgagggagct
1321 gaatatgatg atcagaccag tcaaagggag aaagaagatg ataaagtctt ccctggtgga
1381 agccatacat atgtctggca ggtcctgaaa gagaatggtc caatggcctc tgacccactg
1441 tgccttacct actcatatct ttctcatgtg gacctggtaa aagacttgaa ttcaggcctc
1501 attggagccc tactagtatg tagagaaggg agtctggcca aggaaaagac acagaccttg
1561 cacaaattta tactactttt tgctgtattt gatgaaggga aaagttggca ctcagaaaca
1621 aagaactcct tgatgcagga tagggatgct gcatctgctc gggcctggcc taaaatgcac
1681 acagtcaatg gttatgtaaa caggtctctg ccaggtctga ttggatgcca caggaaatca
1741 gtctattggc atgtgattgg aatgggcacc actcctgaag tgcactcaat attcctcgaa
1801 ggtcacacat ttcttgtgag gaaccatcgc caggcgtcct tggaaatctc gccaataact
1861 ttccttactg ctcaaacact cttgatggac cttggacagt ttctactgtt ttgtcatatc
1921 tcttcccacc aacatgatgg catggaagct tatgtcaaag tagacagctg tccagaggaa
1981 ccccaactac gaatgaaaaa taatgaagaa gcggaagact atgatgatga tcttactgat
2041 tctgaaatgg atgtggtcag gtttgatgat gacaactctc cttcctttat ccaaattcgc
2101 tcagttgcca agaagcatcc taaaacttgg gtacattaca ttgctgctga agaggaggac
2161 tgggactatg ctcccttagt cctcgccccc gatgacagaa gttataaaag tcaatatttg
2221 aacaatggcc ctcagcggat tggtaggaag tacaaaaaag tccgatttat ggcatacaca
2281 gatgaaacct ttaagactcg tgaagctatt cagcatgaat caggaatctt gggaccttta
2341 ctttatgggg aagttggaga cacactgttg attatattta agaatcaagc aagcagacca
2401 tataacatct accctcacgg aatcactgat gtccgtcctt tgtattcaag gagattacca
2461 aaaggtgtaa aacatttgaa ggattttcca attctgccag gagaaatatt caaatataaa
2521 tggacagtga ctgtagaaga tgggccaact aaatcagatc ctcggtgcct gacccgctat
2581 tactctagtt tcgttaatat ggagagagat ctagcttcag gactcattgg ccctctcctc
2641 atctgctaca aagaatctgt agatcaaaga ggaaaccaga taatgtcaga caagaggaat
2701 gtcatcctgt tttctgtatt tgatgagaac cgaagctggt acctcacaga gaatatacaa
2761 cgctttctcc ccaatccagc tggagtgcag cttgaggatc cagagttcca agcctccaac
2821 atcatgcaca gcatcaatgg ctatgttttt gatagtttgc agttgtcagt ttgtttgcat
2881 gaggtggcat actggtacat tctaagcatt ggagcacaga ctgacttcct ttctgtcttc
2941 ttctctggat ataccttcaa acacaaaatg gtctatgaag acacactcac cctattccca
3001 ttctcaggag aaactgtctt catgtcgatg gaaaacccag gtctatggat tctggggtgc
3061 cacaactcag actttcggaa cagaggcatg accgccttac tgaaggtttc tagttgtgac
3121 aagaacactg gtgattatta cgaggacagt tatgaagata tttcagcata cttgctgagt
3181 aaaaacaatg ccattgaacc aagaagcttc tcccagaatt caagacaccc tagcactagg
3241 caaaagcaat ttaatgccac cacaattcca gaaaatgaca tagagaagac tgacccttgg
3301 tttgcacaca gaacacctat gcctaaaata caaaatgtct cctctagtga tttgttgatg
3361 ctcttgcgac agagtcctac tccacatggg ctatccttat ctgatctcca agaagccaaa
3421 tatgagactt tttctgatga tccatcacct ggagcaatag acagtaataa cagcctgtct
3481 gaaatgacac acttcaggcc acagctccat cacagtgggg acatggtatt tacccctgag
3541 tcaggcctcc aattaagatt aaatgagaaa ctggggacaa ctgcagcaac agagttgaag
3601 aaacttgatt tcaaagtttc tagtacatca aataatctga tttcaacaat tccatcagac
3661 aatttggcag caggtactga taatacaagt tccttaggac ccccaagtat gccagttcat
3721 tatgatagtc aattagatac cactctattt ggcaaaaagt catctcccct tactgagtct
3781 ggtggacctc tgagcttgag tgaagaaaat aatgattcaa agttgttaga atcaggttta
3841 atgaatagcc aagaaagttc atggggaaaa aatgtatcgt caacagagag tggtaggtta
3901 tttaaaggga aaagagctca tggacctgct ttgttgacta aagataatgc cttattcaaa
3961 gttagcatct ctttgttaaa gacaaacaaa acttccaata attcagcaac taatagaaag
4021 actcacattg atggcccatc attattaatt gagaatagtc catcagtctg gcaaaatata
4081 ttagaaagtg acactgagtt taaaaaagtg acacctttga ttcatgacag aatgcttatg
4141 gacaaaaatg ctacagcttt gaggctaaat catatgtcaa ataaaactac ttcatcaaaa
4201 aacatggaaa tggtccaaca gaaaaaagag ggccccattc caccagatgc acaaaatcca
4261 gatatgtcgt tctttaagat gctattcttg ccagaatcag caaggtggat acaaaggact
4321 catggaaaga actctctgaa ctctgggcaa ggccccagtc caaagcaatt agtatcctta
4381 ggaccagaaa aatctgtgga aggtcagaat ttcttgtctg agaaaaacaa agtggtagta
4441 ggaaagggtg aatttacaaa ggacgtagga ctcaaagaga tggtttttcc aagcagcaga
4501 aacctatttc ttactaactt ggataattta catgaaaata atacacacaa tcaagaaaaa
4561 aaaattcagg aagaaataga aaagaaggaa acattaatcc aagagaatgt agttttgcct
4621 cagatacata cagtgactgg cactaagaat ttcatgaaga accttttctt actgagcact
4681 aggcaaaatg tagaaggttc atatgacggg gcatatgctc cagtacttca agattttagg
4741 tcattaaatg attcaacaaa tagaacaaag aaacacacag ctcatttctc aaaaaaaggg
4801 gaggaagaaa acttggaagg cttgggaaat caaaccaagc aaattgtaga gaaatatgca
4861 tgcaccacaa ggatatctcc taatacaagc cagcagaatt ttgtcacgca acgtagtaag
4921 agagctttga aacaattcag actcccacta gaagaaacag aacttgaaaa aaggataatt
4981 gtggatgaca cctcaaccca gtggtccaaa aacatgaaac atttgacccc gagcaccctc
5041 acacagatag actacaatga gaaggagaaa ggggccatta ctcagtctcc cttatcagat
5101 tgccttacga ggagtcatag catccctcaa gcaaatagat ctccattacc cattgcaaag
5161 gtatcatcat ttccatctat tagacctata tatctgacca gggtcctatt ccaagacaac
5221 tcttctcatc ttccagcagc atcttataga aagaaagatt ctggggtcca agaaagcagt
5281 catttcttac aaggagccaa aaaaaataac ctttctttag ccattctaac cttggagatg
5341 actggtgatc aaagagaggt tggctccctg gggacaagtg ccacaaattc agtcacatac
5401 aagaaagttg agaacactgt tctcccgaaa ccagacttgc ccaaaacatc tggcaaagtt
5461 gaattgcttc caaaagttca catttatcag aaggacctat tccctacgga aactagcaat
5521 gggtctcctg gccatctgga tctcgtggaa gggagccttc ttcagggaac agagggagcg
5581 attaagtgga atgaagcaaa cagacctgga aaagttccct ttctgagagt agcaacagaa
5641 agctctgcaa agactccctc caagctattg gatcctcttg cttgggataa ccactatggt
5701 actcagatac caaaagaaga gtggaaatcc caagagaagt caccagaaaa aacagctttt
5761 aagaaaaagg ataccatttt gtccctgaac gcttgtgaaa gcaatcatgc aatagcagca
5821 ataaatgagg gacaaaataa gcccgaaata gaagtcacct gggcaaagca aggtaggact
5881 gaaaggctgt gctctcaaaa cccaccagtc ttgaaacgcc atcaacggga aataactcgt
5941 actactcttc agtcagatca agaggaaatt gactatgatg ataccatatc agttgaaatg
6001 aagaaggaag attttgacat ttatgatgag gatgaaaatc agagcccccg cagctttcaa
6061 aagaaaacac gacactattt tattgctgca gtggagaggc tctgggatta tgggatgagt
6121 agctccccac atgttctaag aaacagggct cagagtggca gtgtccctca gttcaagaaa
6181 gttgttttcc aggaatttac tgatggctcc tttactcagc ccttataccg tggagaacta
6241 aatgaacatt tgggactcct ggggccatat ataagagcag aagttgaaga taatatcatg
6301 gtaactttca gaaatcaggc ctctcgtccc tattccttct attctagcct tatttcttat
6361 gaggaagatc agaggcaagg agcagaacct agaaaaaact ttgtcaagcc taatgaaacc
6421 aaaacttact tttggaaagt gcaacatcat atggcaccca ctaaagatga gtttgactgc
6481 aaagcctggg cttatttctc tgatgttgac ctggaaaaag atgtgcactc aggcctgatt
6541 ggaccccttc tggtctgcca cactaacaca ctgaaccctg ctcatgggag acaagtgaca
6601 gtacaggaat ttgctctgtt tttcaccatc tttgatgaga ccaaaagctg gtacttcact
6661 gaaaatatgg aaagaaactg cagggctccc tgcaatatcc agatggaaga tcccactttt
6721 aaagagaatt atcgcttcca tgcaatcaat ggctacataa tggatacact acctggctta
6781 gtaatggctc aggatcaaag gattcgatgg tatctgctca gcatgggcag caatgaaaac
6841 atccattcta ttcatttcag tggacatgtg ttcactgtac gaaaaaaaga ggagtataaa
6901 atggcactgt acaatctcta tccaggtgtt tttgagacag tggaaatgtt accatccaaa
6961 gctggaattt ggcgggtgga atgccttatt ggcgagcatc tacatgctgg gatgagcaca
7021 ctttttctgg tgtacagcaa taagtgtcag actcccctgg gaatggcttc tggacacatt
7081 agagattttc agattacagc ttcaggacaa tatggacagt gggccccaaa gctggccaga
7141 cttcattatt ccggatcaat caatgcctgg agcaccaagg agcccttttc ttggatcaag
7201 gtggatctgt tggcaccaat gattattcac ggcatcaaga cccagggtgc ccgtcagaag
7261 ttctccagcc tctacatctc tcagtttatc atcatgtata gtcttgatgg gaagaagtgg
7321 cagacttatc gaggaaattc cactggaacc ttaatggtct tctttggcaa tgtggattca
7381 tctgggataa aacacaatat ttttaaccct ccaattattg ctcgatacat ccgtttgcac
7441 ccaactcatt atagcattcg cagcactctt cgcatggagt tgatgggctg tgatttaaat
7501 agttgcagca tgccattggg aatggagagt aaagcaatat cagatgcaca gattactgct
7561 tcatcctact ttaccaatat gtttgccacc tggtctcctt caaaagctcg acttcacctc
7621 caagggagga gtaatgcctg gagacctcag gtgaataatc caaaagagtg gctgcaagtg
7681 gacttccaga agacaatgaa agtcacagga gtaactactc agggagtaaa atctctgctt
7741 accagcatgt atgtgaagga gttcctcatc tccagcagtc aagatggcca tcagtggact
7801 ctcttttttc agaatggcaa agtaaaggtt tttcagggaa atcaagactc cttcacacct
7861 gtggtgaact ctctagaccc accgttactg actcgctacc ttcgaattca cccccagagt
7921 tgggtgcacc agattgccct gaggatggag gttctgggct gcgaggcaca ggacctctac
7981 tgagcggccg ctcgaggtca cccattcgaa caaaaactca tctcagaaga ggatctgaat
8041 atgcataccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact
8101 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
8161 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
8221 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
8281 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
8341 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
8401 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
8461 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
8521 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
8581 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
8641 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
8701 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta
8761 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
8821 tagggtgtgg aaagtcccca ggctccccag gcaggcagaa gtatgcaaag catgcatctc
8881 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
8941 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
9001 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
9061 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
9121 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
9181 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
9241 tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
9301 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
9361 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
9421 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
9481 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
9541 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
9601 tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg
9661 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
9721 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
9781 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
9841 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
9901 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
9961 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
10021 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
10081 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
10141 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
10201 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
10261 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
10321 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
10381 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
10441 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
10501 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
10561 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
10621 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
10681 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
10741 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
10801 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
10861 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
10921 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
10981 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
11041 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
11101 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
11161 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
11221 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
11281 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
11341 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
11401 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
11461 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
11521 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
11581 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
11641 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
11701 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
11761 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
11821 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
11881 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
11941 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
12001 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
12061 ttccccgaaa agtgccacct gacgtc
//
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：pCDNA4-F8人源基因质粒
别称: NM_000132.3;AHF;F8B;F8C;HEMA;FVIII;D
启动子：CMV
复制子: pUC
原核抗性: Amp
筛选标记：Zeo
克隆菌株: DH5a
培养条件: 37度
质粒属性
载体宿主：哺乳细胞
载体用途：蛋白表达
基因种属：人
基因类型：ORF
原核抗性：Amp
真核抗性：Zeo
荧光蛋白：
质粒简介
pCDNA3.0-HA-p53质粒由上海烜雅生物科技有限公司提供
质粒图谱
质粒序列
LOCUS Exported 12086 bp ds-DNA circular SYN 21-JAN-2018
FEATURES Location/Qualifiers
source 1..12086
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 928..7983
/codon_start=1
/label=F8
/translation="MQIELSTCFFLCLLRFCFSATRRYYLGAVELSWDYMQSDLGELPV
DARFPPRVPKSFPFNTSVVYKKTLFVEFTDHLFNIAKPRPPWMGLLGPTIQAEVYDTVV
ITLKNMASHPVSLHAVGVSYWKASEGAEYDDQTSQREKEDDKVFPGGSHTYVWQVLKEN
GPM-ASDPLCLTYSYLSHVDLVKDLNSGLIGALLVCREGSLAKEKTQTLHKFILLFAVFD
EGKSWHSETKNSLMQDRDAASARAWPKMHTVNGYVNRSLPGLIGCHRKSVYWHVIGMGT
TPEVHSIFLEGHTFLVRNHRQASLEISPITFLTAQTLLMDLGQFLLFCHISSHQHDGME
AYVKVDSCPEEPQLRMKNNEEAEDYDDDLTDSEMDVVRFDDDNSPSFIQIRSVAKKHPK
TWVHYIAAEEEDWDYAPLVLAPDDRSYKSQYLNNGPQRIGRKYKKVRFMAYTDETFKTR
EAIQHESGILGPLLYGEVGDTLLIIFKNQASRPYNIYPHGITDVRPLYSRRLPKGVKHL
KDFPILPGEIFKYKWTVTVEDGPTKSDPRCLTRYYSSFVNMERDLASGLIGPLLICYKE
SVDQRGNQIMSDKRNVILFSVFDENRSWYLTENIQRFLPNPAGVQLEDPEFQASNIMHS
INGYVFDSLQLSVCLHEVAYWYILSIGAQTDFLSVFFSGYTFKHKMVYEDTLTLFPFSG
ETVFMSMENPGLWILGCHNSDFRNRGMTALLKVSSCDKNTGDYYEDSYEDISAYLLSKN
NAIEPRSFSQNSRHPSTRQKQFNATTIPENDIEKTDPWFAHRTPMPKIQNVSSSDLLML
LRQSPTPHGLSL-SDLQEAKYETFSDDPSPGAIDSNNSLSEMTHFRPQLHHSGDMVFTPE
SGLQLRLNEKLGTTAATELKKLDFKVSSTSNNLISTIPSDNLAAGTDNTSSLGPPSMPV
HYDSQLDTTLFGKKSSPLTESGGPLSLSEENNDSKLLESGLMNSQESSWGKNVSSTESG
RLFKGKRAHGPALLTKDNALFKVSISLLKTNKTSNNSATNRKTHIDGPSLLIENSPSVW
QNILESDTEFKKVTPLIHDRMLMDKNATALRLNHMSNKTTSSKNMEMVQQKKEGPIPPD
AQNPDMSFFKMLFLPESARWIQRTHGKNSLNSGQGPSPKQLVSLGPEKSVEGQNFLSEK
NKVVVGKGEFTKDVGLKEMVFPSSRNLFLTNLDNLHENNTHNQEKKIQEEIEKKETLIQ
ENVVLPQIHTVTGTKNFMKNLFLLSTRQNVEGSYDGAYAPVLQDFRSLNDSTNRTKKHT
AHFSKKGEEENLEGLGNQTKQIVEKYACTTRISPNTSQQNFVTQRSKRALKQFRLPLEE
TELEKRIIVDDTSTQWSKNMKHLTPSTLTQIDYNEKEKGAITQSPL-SDCLTRSHSIPQA
NRSPLPIAKVSSFPSIRPIYLTRVLFQDNSSHLPAASYRKKDSGVQESSHFLQGAKKNN
LSLAILTLEMTGDQREVGSLGTSATNSVTYKKVENTVLPKPDLPKTSGKVELLPKVHIY
QKDLFPTETSNGSPGHLDLVEGSLLQGTEGAIKWNEANRPGKVPFLRVATESSAKTPSK
LLDPLAWDNHYGTQIPKEEWKSQEKSPEKTAFKKKDTILSLNACESNHAIAAINEGQNK
PEIEVTWAKQGRTERLCSQNPPVLKRHQREITRTTLQSDQEEIDYDDTISVEMKKEDFD
IYDEDENQSPRSFQKKTRHYFIAAVERLWDYGMSSSPHVLRNRAQSGSVPQFKKVVFQE
FTDGSFTQPLYRGELNEHLGLLGPYIRAEVEDNIMVTFRNQASRPYSFYSSLISYEEDQ
RQGAEPRKNFVKPNETKTYFWKVQHHMAPTKDEFDCKAWAYFSDVDLEKDVHSGLIGPL
LVCHTNTLNPAHGRQVTVQEFALFFTIFDETKSWYFTENMERNCRAPCNIQMEDPTFKE
NYRFHAINGYIMDTLPGLVMAQDQRIRWYLLSMGSNENIHSIHFSGHVFTVRKKEEYKM
ALYNLYPGVFETVEMLPSKAGIWRVECLIGEHLHAGMSTLFLVYSNKCQTPLGMASGHI
RDFQITASGQYGQWAPKLARLHYSGSINAWSTKEPFSWIKVDLLAPMIIHGIKTQGARQ
KFSSLYISQFIIMYSLDGKKWQTYRGNSTGTLMVFFGNVDSSGIKHNIFNPPIIARYIR
LHPTHYSIRSTLRMELMGCDLNSCSMPLGMESKAISDAQITASSYFTNMFATWSPSKAR
LHLQGRSNAWRPQVNNPKEWLQVDFQKTMKVTGVTTQGVKSLLTSMYVKEFLISSSQDG
HQWTLFFQNGKVKVFQGNQDSFTPVVNSLDPPLLTRYLRIHPQSWVHQIALRMEVLGCE
AQDLY"
polyA_signal 8099..8323
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 8369..8797
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 8811..9141
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 9189..9236
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 9255..9629
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=Zeo
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 9759..9880
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(10331..10919)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(11090..11950)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(11951..12055)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggaacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
901 taagcttcgc gacgtacggc cgccaccatg caaatagagc tctccacctg cttctttctg
961 tgccttttgc gattctgctt tagtgccacc agaagatact acctgggtgc agtggaactg
1021 tcatgggact atatgcaaag tgatctcggt gagctgcctg tggacgcaag atttcctcct
1081 agagtgccaa aatcttttcc attcaacacc tcagtcgtgt acaaaaagac tctgtttgta
1141 gaattcacgg atcacctttt caacatcgct aagccaaggc caccctggat gggtctgcta
1201 ggtcctacca tccaggctga ggtttatgat acagtggtca ttacacttaa gaacatggct
1261 tcccatcctg tcagtcttca tgctgttggt gtatcctact ggaaagcttc tgagggagct
1321 gaatatgatg atcagaccag tcaaagggag aaagaagatg ataaagtctt ccctggtgga
1381 agccatacat atgtctggca ggtcctgaaa gagaatggtc caatggcctc tgacccactg
1441 tgccttacct actcatatct ttctcatgtg gacctggtaa aagacttgaa ttcaggcctc
1501 attggagccc tactagtatg tagagaaggg agtctggcca aggaaaagac acagaccttg
1561 cacaaattta tactactttt tgctgtattt gatgaaggga aaagttggca ctcagaaaca
1621 aagaactcct tgatgcagga tagggatgct gcatctgctc gggcctggcc taaaatgcac
1681 acagtcaatg gttatgtaaa caggtctctg ccaggtctga ttggatgcca caggaaatca
1741 gtctattggc atgtgattgg aatgggcacc actcctgaag tgcactcaat attcctcgaa
1801 ggtcacacat ttcttgtgag gaaccatcgc caggcgtcct tggaaatctc gccaataact
1861 ttccttactg ctcaaacact cttgatggac cttggacagt ttctactgtt ttgtcatatc
1921 tcttcccacc aacatgatgg catggaagct tatgtcaaag tagacagctg tccagaggaa
1981 ccccaactac gaatgaaaaa taatgaagaa gcggaagact atgatgatga tcttactgat
2041 tctgaaatgg atgtggtcag gtttgatgat gacaactctc cttcctttat ccaaattcgc
2101 tcagttgcca agaagcatcc taaaacttgg gtacattaca ttgctgctga agaggaggac
2161 tgggactatg ctcccttagt cctcgccccc gatgacagaa gttataaaag tcaatatttg
2221 aacaatggcc ctcagcggat tggtaggaag tacaaaaaag tccgatttat ggcatacaca
2281 gatgaaacct ttaagactcg tgaagctatt cagcatgaat caggaatctt gggaccttta
2341 ctttatgggg aagttggaga cacactgttg attatattta agaatcaagc aagcagacca
2401 tataacatct accctcacgg aatcactgat gtccgtcctt tgtattcaag gagattacca
2461 aaaggtgtaa aacatttgaa ggattttcca attctgccag gagaaatatt caaatataaa
2521 tggacagtga ctgtagaaga tgggccaact aaatcagatc ctcggtgcct gacccgctat
2581 tactctagtt tcgttaatat ggagagagat ctagcttcag gactcattgg ccctctcctc
2641 atctgctaca aagaatctgt agatcaaaga ggaaaccaga taatgtcaga caagaggaat
2701 gtcatcctgt tttctgtatt tgatgagaac cgaagctggt acctcacaga gaatatacaa
2761 cgctttctcc ccaatccagc tggagtgcag cttgaggatc cagagttcca agcctccaac
2821 atcatgcaca gcatcaatgg ctatgttttt gatagtttgc agttgtcagt ttgtttgcat
2881 gaggtggcat actggtacat tctaagcatt ggagcacaga ctgacttcct ttctgtcttc
2941 ttctctggat ataccttcaa acacaaaatg gtctatgaag acacactcac cctattccca
3001 ttctcaggag aaactgtctt catgtcgatg gaaaacccag gtctatggat tctggggtgc
3061 cacaactcag actttcggaa cagaggcatg accgccttac tgaaggtttc tagttgtgac
3121 aagaacactg gtgattatta cgaggacagt tatgaagata tttcagcata cttgctgagt
3181 aaaaacaatg ccattgaacc aagaagcttc tcccagaatt caagacaccc tagcactagg
3241 caaaagcaat ttaatgccac cacaattcca gaaaatgaca tagagaagac tgacccttgg
3301 tttgcacaca gaacacctat gcctaaaata caaaatgtct cctctagtga tttgttgatg
3361 ctcttgcgac agagtcctac tccacatggg ctatccttat ctgatctcca agaagccaaa
3421 tatgagactt tttctgatga tccatcacct ggagcaatag acagtaataa cagcctgtct
3481 gaaatgacac acttcaggcc acagctccat cacagtgggg acatggtatt tacccctgag
3541 tcaggcctcc aattaagatt aaatgagaaa ctggggacaa ctgcagcaac agagttgaag
3601 aaacttgatt tcaaagtttc tagtacatca aataatctga tttcaacaat tccatcagac
3661 aatttggcag caggtactga taatacaagt tccttaggac ccccaagtat gccagttcat
3721 tatgatagtc aattagatac cactctattt ggcaaaaagt catctcccct tactgagtct
3781 ggtggacctc tgagcttgag tgaagaaaat aatgattcaa agttgttaga atcaggttta
3841 atgaatagcc aagaaagttc atggggaaaa aatgtatcgt caacagagag tggtaggtta
3901 tttaaaggga aaagagctca tggacctgct ttgttgacta aagataatgc cttattcaaa
3961 gttagcatct ctttgttaaa gacaaacaaa acttccaata attcagcaac taatagaaag
4021 actcacattg atggcccatc attattaatt gagaatagtc catcagtctg gcaaaatata
4081 ttagaaagtg acactgagtt taaaaaagtg acacctttga ttcatgacag aatgcttatg
4141 gacaaaaatg ctacagcttt gaggctaaat catatgtcaa ataaaactac ttcatcaaaa
4201 aacatggaaa tggtccaaca gaaaaaagag ggccccattc caccagatgc acaaaatcca
4261 gatatgtcgt tctttaagat gctattcttg ccagaatcag caaggtggat acaaaggact
4321 catggaaaga actctctgaa ctctgggcaa ggccccagtc caaagcaatt agtatcctta
4381 ggaccagaaa aatctgtgga aggtcagaat ttcttgtctg agaaaaacaa agtggtagta
4441 ggaaagggtg aatttacaaa ggacgtagga ctcaaagaga tggtttttcc aagcagcaga
4501 aacctatttc ttactaactt ggataattta catgaaaata atacacacaa tcaagaaaaa
4561 aaaattcagg aagaaataga aaagaaggaa acattaatcc aagagaatgt agttttgcct
4621 cagatacata cagtgactgg cactaagaat ttcatgaaga accttttctt actgagcact
4681 aggcaaaatg tagaaggttc atatgacggg gcatatgctc cagtacttca agattttagg
4741 tcattaaatg attcaacaaa tagaacaaag aaacacacag ctcatttctc aaaaaaaggg
4801 gaggaagaaa acttggaagg cttgggaaat caaaccaagc aaattgtaga gaaatatgca
4861 tgcaccacaa ggatatctcc taatacaagc cagcagaatt ttgtcacgca acgtagtaag
4921 agagctttga aacaattcag actcccacta gaagaaacag aacttgaaaa aaggataatt
4981 gtggatgaca cctcaaccca gtggtccaaa aacatgaaac atttgacccc gagcaccctc
5041 acacagatag actacaatga gaaggagaaa ggggccatta ctcagtctcc cttatcagat
5101 tgccttacga ggagtcatag catccctcaa gcaaatagat ctccattacc cattgcaaag
5161 gtatcatcat ttccatctat tagacctata tatctgacca gggtcctatt ccaagacaac
5221 tcttctcatc ttccagcagc atcttataga aagaaagatt ctggggtcca agaaagcagt
5281 catttcttac aaggagccaa aaaaaataac ctttctttag ccattctaac cttggagatg
5341 actggtgatc aaagagaggt tggctccctg gggacaagtg ccacaaattc agtcacatac
5401 aagaaagttg agaacactgt tctcccgaaa ccagacttgc ccaaaacatc tggcaaagtt
5461 gaattgcttc caaaagttca catttatcag aaggacctat tccctacgga aactagcaat
5521 gggtctcctg gccatctgga tctcgtggaa gggagccttc ttcagggaac agagggagcg
5581 attaagtgga atgaagcaaa cagacctgga aaagttccct ttctgagagt agcaacagaa
5641 agctctgcaa agactccctc caagctattg gatcctcttg cttgggataa ccactatggt
5701 actcagatac caaaagaaga gtggaaatcc caagagaagt caccagaaaa aacagctttt
5761 aagaaaaagg ataccatttt gtccctgaac gcttgtgaaa gcaatcatgc aatagcagca
5821 ataaatgagg gacaaaataa gcccgaaata gaagtcacct gggcaaagca aggtaggact
5881 gaaaggctgt gctctcaaaa cccaccagtc ttgaaacgcc atcaacggga aataactcgt
5941 actactcttc agtcagatca agaggaaatt gactatgatg ataccatatc agttgaaatg
6001 aagaaggaag attttgacat ttatgatgag gatgaaaatc agagcccccg cagctttcaa
6061 aagaaaacac gacactattt tattgctgca gtggagaggc tctgggatta tgggatgagt
6121 agctccccac atgttctaag aaacagggct cagagtggca gtgtccctca gttcaagaaa
6181 gttgttttcc aggaatttac tgatggctcc tttactcagc ccttataccg tggagaacta
6241 aatgaacatt tgggactcct ggggccatat ataagagcag aagttgaaga taatatcatg
6301 gtaactttca gaaatcaggc ctctcgtccc tattccttct attctagcct tatttcttat
6361 gaggaagatc agaggcaagg agcagaacct agaaaaaact ttgtcaagcc taatgaaacc
6421 aaaacttact tttggaaagt gcaacatcat atggcaccca ctaaagatga gtttgactgc
6481 aaagcctggg cttatttctc tgatgttgac ctggaaaaag atgtgcactc aggcctgatt
6541 ggaccccttc tggtctgcca cactaacaca ctgaaccctg ctcatgggag acaagtgaca
6601 gtacaggaat ttgctctgtt tttcaccatc tttgatgaga ccaaaagctg gtacttcact
6661 gaaaatatgg aaagaaactg cagggctccc tgcaatatcc agatggaaga tcccactttt
6721 aaagagaatt atcgcttcca tgcaatcaat ggctacataa tggatacact acctggctta
6781 gtaatggctc aggatcaaag gattcgatgg tatctgctca gcatgggcag caatgaaaac
6841 atccattcta ttcatttcag tggacatgtg ttcactgtac gaaaaaaaga ggagtataaa
6901 atggcactgt acaatctcta tccaggtgtt tttgagacag tggaaatgtt accatccaaa
6961 gctggaattt ggcgggtgga atgccttatt ggcgagcatc tacatgctgg gatgagcaca
7021 ctttttctgg tgtacagcaa taagtgtcag actcccctgg gaatggcttc tggacacatt
7081 agagattttc agattacagc ttcaggacaa tatggacagt gggccccaaa gctggccaga
7141 cttcattatt ccggatcaat caatgcctgg agcaccaagg agcccttttc ttggatcaag
7201 gtggatctgt tggcaccaat gattattcac ggcatcaaga cccagggtgc ccgtcagaag
7261 ttctccagcc tctacatctc tcagtttatc atcatgtata gtcttgatgg gaagaagtgg
7321 cagacttatc gaggaaattc cactggaacc ttaatggtct tctttggcaa tgtggattca
7381 tctgggataa aacacaatat ttttaaccct ccaattattg ctcgatacat ccgtttgcac
7441 ccaactcatt atagcattcg cagcactctt cgcatggagt tgatgggctg tgatttaaat
7501 agttgcagca tgccattggg aatggagagt aaagcaatat cagatgcaca gattactgct
7561 tcatcctact ttaccaatat gtttgccacc tggtctcctt caaaagctcg acttcacctc
7621 caagggagga gtaatgcctg gagacctcag gtgaataatc caaaagagtg gctgcaagtg
7681 gacttccaga agacaatgaa agtcacagga gtaactactc agggagtaaa atctctgctt
7741 accagcatgt atgtgaagga gttcctcatc tccagcagtc aagatggcca tcagtggact
7801 ctcttttttc agaatggcaa agtaaaggtt tttcagggaa atcaagactc cttcacacct
7861 gtggtgaact ctctagaccc accgttactg actcgctacc ttcgaattca cccccagagt
7921 tgggtgcacc agattgccct gaggatggag gttctgggct gcgaggcaca ggacctctac
7981 tgagcggccg ctcgaggtca cccattcgaa caaaaactca tctcagaaga ggatctgaat
8041 atgcataccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact
8101 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
8161 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
8221 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
8281 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
8341 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
8401 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
8461 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
8521 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
8581 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
8641 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
8701 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta
8761 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
8821 tagggtgtgg aaagtcccca ggctccccag gcaggcagaa gtatgcaaag catgcatctc
8881 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
8941 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
9001 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
9061 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
9121 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
9181 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
9241 tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
9301 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
9361 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
9421 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
9481 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
9541 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
9601 tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg
9661 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
9721 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
9781 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
9841 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
9901 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
9961 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
10021 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
10081 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
10141 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
10201 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
10261 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
10321 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
10381 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
10441 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
10501 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
10561 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
10621 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
10681 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
10741 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
10801 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
10861 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
10921 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
10981 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
11041 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
11101 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
11161 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
11221 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
11281 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
11341 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
11401 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
11461 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
11521 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
11581 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
11641 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
11701 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
11761 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
11821 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
11881 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
11941 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
12001 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
12061 ttccccgaaa agtgccacct gacgtc
//
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
| P3822/pCMV-SPORT6-GALNT15（1同义突变） |
| P3823/pCMV-SPORT6-ZC2HC1A人源基因质粒 |
| P0637/pCR4-MME人源基因质粒 |
| P0645/pCR4-IL12A人源基因质粒 |
| P0646/pCR4-PKD2人源基因质粒 |
| P4957/pCR-XL-TOPO-EFCAB4B人源基因质粒 |
| P0647/pDNR-Dual-IL12B人源基因质粒 |
| P4822/pDNR-LIB-PABPN1（2点突变）人源基因质粒 |
| P2411/pOTB7-LILRB1人源基因质粒 |
| P4943/pOTB7-PDIA4（1同义突变）人源基因质粒 |
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文献和实验相关实验
【求助】MDCK转染与免疫荧光问题，解决问题追加丁当，谢谢！
wen-wen 实验组：将带有c-myc和UT-B的pcDNA3.0质粒转染到MDCK细胞当中 对照组：1、将带有c-Myc和AQP1的pcDNA3.0质粒转染到MDCK细胞； 2、将GFP的质粒（就是一个只有GFP的质粒，没有UT-B,AQP1）转染MDCK细胞中，主要目的是想看我的操作技术怎么样； 3、过表达AQP1-MDCK的稳转细胞系。 然后通过免疫荧光来证实转染结果。（因为质粒若带有GFP会影响
表达载体的构建方法及步骤 一、载体的选择及如何阅读质粒图谱 目前，载体主要有病毒和非病毒两大类,其中质粒 DNA 是一种新的非病毒转基因载体。 一个合格质粒的组成要素： （1）复制起始位点 Ori 即控制复制起始的位点。原核生物 DNA 分子中只有一个复制起始点。而 真核生物 DNA 分子有多个复制起始位点。 （2）抗生素抗性基因可以便于加以检测，如 Amp+ ,Kan+ （3）多克隆位点 MCS 克隆携带外源基因片段 （4） P/E 启动子/增强
zhangyuxianggx 各位大侠好！现有pcDNA3-mCherry和pGEM-hgene两个东西在滤纸上，老师就说让构建载体，具体是什么意思啊？我理解的是一个真核表达载体和构建好的pGEM质粒，但是不知下一步该怎么做？恳请各位好心的大侠指点一二。 shylook 直接插入cDNA吧 zhangyuxianggx 那个pGEM就是已经插入了DNA的载体。请问这两个东西溶下来以后
pCDNA4-F8人源基因质粒
￥100 - 1000
