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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：
pCXUN-FLAG
- 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：pCXUN-FLAG植物表达质粒
别称: pCXUN-FLAG
质粒属性
质粒简介
pCXUN-FLAG是一个植物pCX系列蛋白表达质粒。
质粒图谱
质粒序列
LOCUS Exported 12035 bp ds-DNA circular SYN 04-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..12035
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 115..367
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS 728..1033
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLL-SDK
VSRELYPVVHIGDESWRMMTTD-MASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
CDS complement(1089..1112)
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
promoter complement(1128..3118)
/label=Ubi promoter
/note="maize polyubiquitin gene promoter"
misc_feature 3359..3383
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 4682..5311
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 5745..6812
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
MRYRNLIEGEASAGS"
rep_origin 6878..7072
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 7416..7556
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(7742..8330)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8417..9211)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 9636..9660
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 9738..9912
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(9952..10977)
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMD-AIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
K"
promoter complement(11044..11721)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
ORIGIN
1 gaattcgagc tcggtacccc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
61 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tcccgatcta
121 gtaacataga tgacaccgcg cgcgataatt tatcctagtt tgcgcgctat attttgtttt
181 ctatcgcgta ttaaatgtat aattgcggga ctctaatcat aaaaacccat ctcataaata
241 acgtcatgca ttacatgtta attattacat gcttaacgta attcaacaga aattatatga
301 taatcatcgc aagaccggca acaggattca atcttaagaa actttattgc caaatgtttg
361 aacgatcggg gaaattcgga tccccaatac ttgtatgggg cttactaaaa gccagataac
421 agtatgcgta tttgcgcgct gatttttgcg gtataagaat atatactgat atgtataccc
481 gaagtatgtc aaaaagaggt atgctatgaa gcagcgtatt acagtgacag ttgacagcga
541 cagctatcag ttgctcaagg catatatgat gtcaatatct ccggtctggt aagcacaacc
601 atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg
661 atggctgagg tcgcccggtt tattgaaatg aacggctctt ttgctgacga gaacaggggc
721 tggtgaaatg cagtttaagg tttacaccta taaaagagag agccgttatc gtctgtttgt
781 ggatgtacag agtgatatta ttgacacgcc cgggcgacgg atggtgatcc ccctggccag
841 tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc atatcgggga
901 tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtctccg ttatcgggga
961 agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta acctgatgtt
1021 ctggggaata taaatgtcag gctcccttat acacagccag tctgcaggtc gacccataca
1081 agtattggct tatcatcatc atccttgtaa tccatggatc ccccgggctg cagaagtaac
1141 accaaacaac agggtgagca tcgacaaaag aaacagtacc aagcaaataa atagcgtatg
1201 aaggcagggc taaaaaaatc cacatatagc tgctgcatat gccatcatcc aagtatatca
1261 agatcaaaat aattataaaa catacttgtt tattataata gataggtact caaggttaga
1321 gcatatgaat agatgctgca tatgccatca tgtatatgca tcagtaaaac ccacatcaac
1381 atgtatacct atcctagatc gatatttcca tccatcttaa actcgtaact atgaagatgt
1441 atgacacaca catacagttc caaaattaat aaatacacca ggtagtttga aacagtattc
1501 tactccgatc tagaacgaat gaacgaccgc ccaaccacac cacatcatca caaccaagcg
1561 aacaaaaagc atctctgtat atgcatcagt aaaacccgca tcaacatgta tacctatcct
1621 agatcgatat ttccatccat catcttcaat tcgtaactat gaatatgtat ggcacacaca
1681 tacagatcca aaattaataa atccaccagg tagtttgaaa cagaattcta ctccgatcta
1741 gaacgaccgc ccaaccagac cacatcatca caaccaagac aaaaaaaagc atgaaaagat
1801 gacccgacaa acaagtgcac ggcatatatt gaaataaagg aaaagggcaa accaaaccct
1861 atgcaacgaa acaaaaaaaa tcatgaaatc gatcccgtct gcggaacggc tagagccatc
1921 ccaggattcc ccaaagagaa acactggcaa gttagcaatc agaacgtgtc tgacgtacag
1981 gtcgcatccg tgtacgaacg ctagcagcac ggatctaaca caaacacgga tctaacacaa
2041 acatgaacag aagtagaact accgggccct aaccatggac cggaacgccg atctagagaa
2101 ggtagagagg gggggggggg gaggacgagc ggcgtacctt gaagcggagg tgccgacggg
2161 tggatttggg ggagatctgg ttgtgtgtgt gtgcgctccg aacaacacga ggttggggaa
2221 agagggtgtg gagggggtgt ctatttatta cggcgggcga ggaagggaaa gcgaaggagc
2281 ggtgggaaag gaatcccccg tagctgccgt gccgtgagag gaggaggagg ccgcctgccg
2341 tgccggctca cgtctgccgc tccgccacgc atttctggat gccgacagcg gagcaagtcc
2401 aacggtggag cggaactctc gagaggggtc cagaggcagc gacagagatg ccgtgccgtc
2461 tgcttcgctt ggcccgacgc gacgctgctg gttcgctggt tggtgtccgt tagactcgtc
2521 gacggcgttt aacaggctgg cattatctac tcgaaacaag aaaaatgttt ccttagtttt
2581 tttaatttct taaagggtat ttgtttaatt tttagtcact ttattttatt ctattttata
2641 tctaaattat taaataaaaa aactaaaata gagttttagt tttcttaatt tagaggctaa
2701 aatagaataa aatagatgta ctaaaaaaat tagtctataa aaaccattaa ccctaaaccc
2761 taaatggatg tactaataaa atggatgaag tattatatag gtgaagctat ttgcaaaaaa
2821 aaaggagaac acatgcacac taaaaagata aaactgtaga gtcctgttgt caaaatactc
2881 aattgtcctt tagaccatgt ctaactgttc atttatatga ttctctaaaa cactgatatt
2941 attgtagtac tatagattat attattcgta gagtaaagtt taaatatatg tataaagata
3001 gataaactgc acttcaaaca agtgtgacaa aaaaaatatg tggtaatttt ttataactta
3061 gacatgcaat gctcattatc tctagagagg ggcacgaccg ggtcacgctg cactgcagga
3121 attcgatatc aagcttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
3181 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
3241 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgctaga
3301 gcagcttgag cttggatcag attgtcgttt cccgccttca gtttaaacta tcagtgtttg
3361 acaggatata ttggcgggta aacctaagag aaaagagcgt ttattagaat aacggatatt
3421 taaaagggcg tgaaaaggtt tatccgttcg tccatttgta tgtgcatgcc aaccacaggg
3481 ttcccctcgg gatcaaagta ctttgatcca acccctccgc tgctatagtg cagtcggctt
3541 ctgacgttca gtgcagccgt cttctgaaaa cgacatgtcg cacaagtcct aagttacgcg
3601 acaggctgcc gccctgccct tttcctggcg ttttcttgtc gcgtgtttta gtcgcataaa
3661 gtagaatact tgcgactaga accggagaca ttacgccatg aacaagagcg ccgccgctgg
3721 cctgctgggc tatgcccgcg tcagcaccga cgaccaggac ttgaccaacc aacgggccga
3781 actgcacgcg gccggctgca ccaagctgtt ttccgagaag atcaccggca ccaggcgcga
3841 ccgcccggag ctggccagga tgcttgacca cctagccctg gcgacgttgt gacagtgacc
3901 aggctagacc gcctggcccg cagcacccgc gacctactgg acattgccga gcgcatccag
3961 gaggccggcg cgggcctgcg tagcctggca gagccgtggg ccgacaccac cacgccggcc
4021 ggccgcatgg tgttgaccgt gttcgccggc attgccgagt tcgagcgttc cctaatcatc
4081 gaccgcaccc ggagcgggcg cgaggccgcc aaggcccgag gcgtgaagtt tggcccccgc
4141 cctaccctca ccccggcaca gatcgcgcac gcccgcgagc tgatcgacca ggaaggccgc
4201 accgtgaaag aggcggctgc actgcttggc gtgcatcgct cgaccctgta ccgcgcactt
4261 gagcgcagcg aggaagtgac gcccaccgag gccaggcggc gcggtgcctt ccgtgaggac
4321 gcattgaccg aggccgacgc cctggcggcc gccgagaatg aacgccaaga ggaacaagca
4381 tgaaaccgca ccaggacggc caggacgaac cgtttttcat taccgaagag atcgaggcgg
4441 agatgatcgc ggccgggtac gtgttcgagc cgcccgcgca cgtctcaacc gtgcggctgc
4501 atgaaatcct ggccggtttg tctgatgcca agctggcggc ctggccggcc agcttggccg
4561 ctgaagaaac cgagcgccgc cgtctaaaaa ggtgatgtgt atttgagtaa aacagcttgc
4621 gtcatgcggt cgctgcgtat atgatgcgat gagtaaataa acaaatacgc aaggggaacg
4681 catgaaggtt atcgctgtac ttaaccagaa aggcgggtca ggcaagacga ccatcgcaac
4741 ccatctagcc cgcgccctgc aactcgccgg ggccgatgtt ctgttagtcg attccgatcc
4801 ccagggcagt gcccgcgatt gggcggccgt gcgggaagat caaccgctaa ccgttgtcgg
4861 catcgaccgc ccgacgattg accgcgacgt gaaggccatc ggccggcgcg acttcgtagt
4921 gatcgacgga gcgccccagg cggcggactt ggctgtgtcc gcgatcaagg cagccgactt
4981 cgtgctgatt ccggtgcagc caagccctta cgacatatgg gcaaccgccg acctggtgga
5041 gctggttaag cagcgcattg aggtcacgga tggaaggcta caagcggcct ttgtcgtgtc
5101 gcgggcgatc aaaggcacgc gcatcggcgg tgaggttgcc gaggcgctgg ccgggtacga
5161 gctgcccatt cttgagtccc gtatcacgca gcgcgtgagc tacccaggca ctgccgccgc
5221 cggcacaacc gttcttgaat cagaacccga gggcgacgct gcccgcgagg tccaggcgct
5281 ggccgctgaa attaaatcaa aactcatttg agttaatgag gtaaagagaa aatgagcaaa
5341 agcacaaaca cgctaagtgc cggccgtccg agcgcacgca gcagcaaggc tgcaacgttg
5401 gccagcctgg cagacacgcc agccatgaag cgggtcaact ttcagttgcc ggcggaggat
5461 cacaccaagc tgaagatgta cgcggtacgc caaggcaaga ccattaccga gctgctatct
5521 gaatacatcg cgcagctacc agagtaaatg agcaaatgaa taaatgagta gatgaatttt
5581 agcggctaaa ggaggcggca tggaaaatca agaacaacca ggcaccgacg ccgtggaatg
5641 ccccatgtgt ggaggaacgg gcggttggcc aggcgtaagc ggctgggttg tctgccggcc
5701 ctgcaatggc actggaaccc ccaagcccga ggaatcggcg tgacggtcgc aaaccatccg
5761 gcccggtaca aatcggcgcg gcgctgggtg atgacctggt ggagaagttg aaggccgcgc
5821 aggccgccca gcggcaacgc atcgaggcag aagcacgccc cggtgaatcg tggcaagcgg
5881 ccgctgatcg aatccgcaaa gaatcccggc aaccgccggc agccggtgcg ccgtcgatta
5941 ggaagccgcc caagggcgac gagcaaccag attttttcgt tccgatgctc tatgacgtgg
6001 gcacccgcga tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg aagcgtgacc
6061 gacgagctgg cgaggtgatc cgctacgagc ttccagacgg gcacgtagag gtttccgcag
6121 ggccggccgg catggccagt gtgtgggatt acgacctggt actgatggcg gtttcccatc
6181 taaccgaatc catgaaccga taccgggaag ggaagggaga caagcccggc cgcgtgttcc
6241 gtccacacgt tgcggacgta ctcaagttct gccggcgagc cgatggcgga aagcagaaag
6301 acgacctggt agaaacctgc attcggttaa acaccacgca cgttgccatg cagcgtacga
6361 agaaggccaa gaacggccgc ctggtgacgg tatccgaggg tgaagccttg attagccgct
6421 acaagatcgt aaagagcgaa accgggcggc cggagtacat cgagatcgag ctagctgatt
6481 ggatgtaccg cgagatcaca gaaggcaaga acccggacgt gctgacggtt caccccgatt
6541 actttttgat cgatcccggc atcggccgtt ttctctaccg cctggcacgc cgcgccgcag
6601 gcaaggcaga agccagatgg ttgttcaaga cgatctacga acgcagtggc agcgccggag
6661 agttcaagaa gttctgtttc accgtgcgca agctgatcgg gtcaaatgac ctgccggagt
6721 acgatttgaa ggaggaggcg gggcaggctg gcccgatcct agtcatgcgc taccgcaacc
6781 tgatcgaggg cgaagcatcc gccggttcct aatgtacgga gcagatgcta gggcaaattg
6841 ccctagcagg ggaaaaaggt cgaaaaggtc tctttcctgt ggatagcacg tacattggga
6901 acccaaagcc gtacattggg aaccggaacc cgtacattgg gaacccaaag ccgtacattg
6961 ggaaccggtc acacatgtaa gtgactgata taaaagagaa aaaaggcgat ttttccgcct
7021 aaaactcttt aaaacttatt aaaactctta aaacccgcct ggcctgtgca taactgtctg
7081 gccagcgcac agccgaagag ctgcaaaaag cgcctaccct tcggtcgctg cgctccctac
7141 gccccgccgc ttcgcgtcgg cctatcgcgg ccgctggccg ctcaaaaatg gctggcctac
7201 ggccaggcaa tctaccaggg cgcggacaag ccgcgccgtc gccactcgac cgccggcgcc
7261 cacatcaagg caccctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
7321 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
7381 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
7441 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
7501 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
7561 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
7621 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
7681 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
7741 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
7801 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
7861 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
7921 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
7981 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
8041 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
8101 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
8161 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
8221 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
8281 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
8341 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
8401 tcatgcattc taggtactaa aacaattcat ccagtaaaat ataatatttt attttctccc
8461 aatcaggctt gatccccagt aagtcaaaaa atagctcgac atactgttct tccccgatat
8521 cctccctgat cgaccggacg cagaaggcaa tgtcatacca cttgtccgcc ctgccgcttc
8581 tcccaagatc aataaagcca cttactttgc catctttcac aaagatgttg ctgtctccca
8641 ggtcgccgtg ggaaaagaca agttcctctt cgggcttttc cgtctttaaa aaatcataca
8701 gctcgcgcgg atctttaaat ggagtgtctt cttcccagtt ttcgcaatcc acatcggcca
8761 gatcgttatt cagtaagtaa tccaattcgg ctaagcggct gtctaagcta ttcgtatagg
8821 gacaatccga tatgtcgatg gagtgaaaga gcctgatgca ctccgcatac agctcgataa
8881 tcttttcagg gctttgttca tcttcatact cttccgagca aaggacgcca tcggcctcac
8941 tcatgagcag attgctccag ccatcatgcc gttcaaagtg caggaccttt ggaacaggca
9001 gctttccttc cagccatagc atcatgtcct tttcccgttc aacatcatag gtggtccctt
9061 tataccggct gtccgtcatt tttaaatata ggttttcatt ttctcccacc agcttatata
9121 ccttagcagg agacattcct tccgtatctt ttacgcagcg gtatttttcg atcagttttt
9181 tcaattccgg tgatattctc attttagcca tttattattt ccttcctctt ttctacagta
9241 tttaaagata ccccaagaag ctaattataa caagacgaac tccaattcac tgttccttgc
9301 attctaaaac cttaaatacc agaaaacagc tttttcaaag ttgttttcaa agttggcgta
9361 taacatagta tcgacggagc cgattttgaa accgcggtga tcacaggcag caacgctctg
9421 tcatcgttac aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc
9481 ttagttgccg ttcttccgaa tagcatcggt aacatgagca aagtctgccg ccttacaacg
9541 gctctcccgc tgacgccgtc ccggactgat gggctgcctg tatcgagtgg tgattttgtg
9601 ccgagctgcc ggtcggggag ctgttggctg gctggtggca ggatatattg tggtgtaaac
9661 aaattgacgc ttagacaact taataacaca ttgcggacgt ttttaatgta ctgaattaac
9721 gccgaattaa ttcgggggat ctggatttta gtactggatt ttggttttag gaattagaaa
9781 ttttattgat agaagtattt tacaaataca aatacatact aagggtttct tatatgctca
9841 acacatgagc gaaaccctat aggaacccta attcccttat ctgggaacta ctcacacatt
9901 attatggaga aactcgagct tgtcgatcga cagatccggt cggcatctac tctatttctt
9961 tgccctcgga cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc
10021 atcggtccag acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc
10081 ggatcggacg attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc
10141 aaccaagctc tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagtcgtgg
10201 cgatcctgca agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc
10261 caaccacggc ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc
10321 ctcgctccag tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc
10381 gaaatccgcg tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc
10441 atcgagagcc tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata
10501 cacatgggga tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc
10561 ttgcggtccg aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc
10621 catagcctcc gcgaccggtt gtagaacagc gggcagttcg gtttcaggca ggtcttgcaa
10681 cgtgacaccc tgtgcacggc gggagatgca ataggtcagg ctctcgctaa actccccaat
10741 gtcaagcact tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc
10801 tttgtagaaa ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc
10861 gaagctgaaa gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc
10921 gaacttttcg atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttcatatc
10981 tcattgcccc ccggatctgc gaaagctcga gagagataga tttgtagaga gagactggtg
11041 atttcagcgt gtcctctcca aatgaaatga acttccttat atagaggaag gtcttgcgaa
11101 ggatagtggg attgtgcgtc atcccttacg tcagtggaga tatcacatca atccacttgc
11161 tttgaagacg tggttggaac gtcttctttt tccacgatgc tcctcgtggg tgggggtcca
11221 tctttgggac cactgtcggc agaggcatct tgaacgatag cctttccttt atcgcaatga
11281 tggcatttgt aggtgccacc ttccttttct actgtccttt tgatgaagtg acagatagct
11341 gggcaatgga atccgaggag gtttcccgat attacccttt gttgaaaagt ctcaatagcc
11401 ctttggtctt ctgagactgt atctttgata ttcttggagt agacgagagt gtcgtgctcc
11461 accatgttat cacatcaatc cacttgcttt gaagacgtgg ttggaacgtc ttctttttcc
11521 acgatgctcc tcgtgggtgg gggtccatct ttgggaccac tgtcggcaga ggcatcttga
11581 acgatagcct ttcctttatc gcaatgatgg catttgtagg tgccaccttc cttttctact
11641 gtccttttga tgaagtgaca gatagctggg caatggaatc cgaggaggtt tcccgatatt
11701 accctttgtt gaaaagtctc aatagccctt tggtcttctg agactgtatc tttgatattc
11761 ttggagtaga cgagagtgtc gtgctccacc atgttggcaa gctgctctag ccaatacgca
11821 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
11881 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
11941 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
12001 aatttcacac aggaaacagc tatgaccatg attac
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：pCXUN-FLAG植物表达质粒
别称: pCXUN-FLAG
| 原核抗性: | Kan |
|---|---|
| 筛选标记: | Hyg |
| 克隆菌株: | DB3.1 |
| 培养条件: | 37度 |
质粒属性
| 载体宿主： | 植物 |
|---|---|
| 载体用途： | 蛋白表达 |
| 基因种属： | |
| 基因类型： | ORF |
| 原核抗性： | Kan |
| 筛选标记： | Hyg |
| 荧光蛋白： |
质粒简介
pCXUN-FLAG是一个植物pCX系列蛋白表达质粒。
质粒图谱
质粒序列
LOCUS Exported 12035 bp ds-DNA circular SYN 04-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..12035
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 115..367
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS 728..1033
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLL-SDK
VSRELYPVVHIGDESWRMMTTD-MASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
CDS complement(1089..1112)
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
promoter complement(1128..3118)
/label=Ubi promoter
/note="maize polyubiquitin gene promoter"
misc_feature 3359..3383
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 4682..5311
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 5745..6812
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
MRYRNLIEGEASAGS"
rep_origin 6878..7072
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 7416..7556
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(7742..8330)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8417..9211)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 9636..9660
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 9738..9912
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(9952..10977)
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMD-AIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
K"
promoter complement(11044..11721)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
ORIGIN
1 gaattcgagc tcggtacccc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
61 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tcccgatcta
121 gtaacataga tgacaccgcg cgcgataatt tatcctagtt tgcgcgctat attttgtttt
181 ctatcgcgta ttaaatgtat aattgcggga ctctaatcat aaaaacccat ctcataaata
241 acgtcatgca ttacatgtta attattacat gcttaacgta attcaacaga aattatatga
301 taatcatcgc aagaccggca acaggattca atcttaagaa actttattgc caaatgtttg
361 aacgatcggg gaaattcgga tccccaatac ttgtatgggg cttactaaaa gccagataac
421 agtatgcgta tttgcgcgct gatttttgcg gtataagaat atatactgat atgtataccc
481 gaagtatgtc aaaaagaggt atgctatgaa gcagcgtatt acagtgacag ttgacagcga
541 cagctatcag ttgctcaagg catatatgat gtcaatatct ccggtctggt aagcacaacc
601 atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg
661 atggctgagg tcgcccggtt tattgaaatg aacggctctt ttgctgacga gaacaggggc
721 tggtgaaatg cagtttaagg tttacaccta taaaagagag agccgttatc gtctgtttgt
781 ggatgtacag agtgatatta ttgacacgcc cgggcgacgg atggtgatcc ccctggccag
841 tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc atatcgggga
901 tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtctccg ttatcgggga
961 agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta acctgatgtt
1021 ctggggaata taaatgtcag gctcccttat acacagccag tctgcaggtc gacccataca
1081 agtattggct tatcatcatc atccttgtaa tccatggatc ccccgggctg cagaagtaac
1141 accaaacaac agggtgagca tcgacaaaag aaacagtacc aagcaaataa atagcgtatg
1201 aaggcagggc taaaaaaatc cacatatagc tgctgcatat gccatcatcc aagtatatca
1261 agatcaaaat aattataaaa catacttgtt tattataata gataggtact caaggttaga
1321 gcatatgaat agatgctgca tatgccatca tgtatatgca tcagtaaaac ccacatcaac
1381 atgtatacct atcctagatc gatatttcca tccatcttaa actcgtaact atgaagatgt
1441 atgacacaca catacagttc caaaattaat aaatacacca ggtagtttga aacagtattc
1501 tactccgatc tagaacgaat gaacgaccgc ccaaccacac cacatcatca caaccaagcg
1561 aacaaaaagc atctctgtat atgcatcagt aaaacccgca tcaacatgta tacctatcct
1621 agatcgatat ttccatccat catcttcaat tcgtaactat gaatatgtat ggcacacaca
1681 tacagatcca aaattaataa atccaccagg tagtttgaaa cagaattcta ctccgatcta
1741 gaacgaccgc ccaaccagac cacatcatca caaccaagac aaaaaaaagc atgaaaagat
1801 gacccgacaa acaagtgcac ggcatatatt gaaataaagg aaaagggcaa accaaaccct
1861 atgcaacgaa acaaaaaaaa tcatgaaatc gatcccgtct gcggaacggc tagagccatc
1921 ccaggattcc ccaaagagaa acactggcaa gttagcaatc agaacgtgtc tgacgtacag
1981 gtcgcatccg tgtacgaacg ctagcagcac ggatctaaca caaacacgga tctaacacaa
2041 acatgaacag aagtagaact accgggccct aaccatggac cggaacgccg atctagagaa
2101 ggtagagagg gggggggggg gaggacgagc ggcgtacctt gaagcggagg tgccgacggg
2161 tggatttggg ggagatctgg ttgtgtgtgt gtgcgctccg aacaacacga ggttggggaa
2221 agagggtgtg gagggggtgt ctatttatta cggcgggcga ggaagggaaa gcgaaggagc
2281 ggtgggaaag gaatcccccg tagctgccgt gccgtgagag gaggaggagg ccgcctgccg
2341 tgccggctca cgtctgccgc tccgccacgc atttctggat gccgacagcg gagcaagtcc
2401 aacggtggag cggaactctc gagaggggtc cagaggcagc gacagagatg ccgtgccgtc
2461 tgcttcgctt ggcccgacgc gacgctgctg gttcgctggt tggtgtccgt tagactcgtc
2521 gacggcgttt aacaggctgg cattatctac tcgaaacaag aaaaatgttt ccttagtttt
2581 tttaatttct taaagggtat ttgtttaatt tttagtcact ttattttatt ctattttata
2641 tctaaattat taaataaaaa aactaaaata gagttttagt tttcttaatt tagaggctaa
2701 aatagaataa aatagatgta ctaaaaaaat tagtctataa aaaccattaa ccctaaaccc
2761 taaatggatg tactaataaa atggatgaag tattatatag gtgaagctat ttgcaaaaaa
2821 aaaggagaac acatgcacac taaaaagata aaactgtaga gtcctgttgt caaaatactc
2881 aattgtcctt tagaccatgt ctaactgttc atttatatga ttctctaaaa cactgatatt
2941 attgtagtac tatagattat attattcgta gagtaaagtt taaatatatg tataaagata
3001 gataaactgc acttcaaaca agtgtgacaa aaaaaatatg tggtaatttt ttataactta
3061 gacatgcaat gctcattatc tctagagagg ggcacgaccg ggtcacgctg cactgcagga
3121 attcgatatc aagcttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
3181 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
3241 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgctaga
3301 gcagcttgag cttggatcag attgtcgttt cccgccttca gtttaaacta tcagtgtttg
3361 acaggatata ttggcgggta aacctaagag aaaagagcgt ttattagaat aacggatatt
3421 taaaagggcg tgaaaaggtt tatccgttcg tccatttgta tgtgcatgcc aaccacaggg
3481 ttcccctcgg gatcaaagta ctttgatcca acccctccgc tgctatagtg cagtcggctt
3541 ctgacgttca gtgcagccgt cttctgaaaa cgacatgtcg cacaagtcct aagttacgcg
3601 acaggctgcc gccctgccct tttcctggcg ttttcttgtc gcgtgtttta gtcgcataaa
3661 gtagaatact tgcgactaga accggagaca ttacgccatg aacaagagcg ccgccgctgg
3721 cctgctgggc tatgcccgcg tcagcaccga cgaccaggac ttgaccaacc aacgggccga
3781 actgcacgcg gccggctgca ccaagctgtt ttccgagaag atcaccggca ccaggcgcga
3841 ccgcccggag ctggccagga tgcttgacca cctagccctg gcgacgttgt gacagtgacc
3901 aggctagacc gcctggcccg cagcacccgc gacctactgg acattgccga gcgcatccag
3961 gaggccggcg cgggcctgcg tagcctggca gagccgtggg ccgacaccac cacgccggcc
4021 ggccgcatgg tgttgaccgt gttcgccggc attgccgagt tcgagcgttc cctaatcatc
4081 gaccgcaccc ggagcgggcg cgaggccgcc aaggcccgag gcgtgaagtt tggcccccgc
4141 cctaccctca ccccggcaca gatcgcgcac gcccgcgagc tgatcgacca ggaaggccgc
4201 accgtgaaag aggcggctgc actgcttggc gtgcatcgct cgaccctgta ccgcgcactt
4261 gagcgcagcg aggaagtgac gcccaccgag gccaggcggc gcggtgcctt ccgtgaggac
4321 gcattgaccg aggccgacgc cctggcggcc gccgagaatg aacgccaaga ggaacaagca
4381 tgaaaccgca ccaggacggc caggacgaac cgtttttcat taccgaagag atcgaggcgg
4441 agatgatcgc ggccgggtac gtgttcgagc cgcccgcgca cgtctcaacc gtgcggctgc
4501 atgaaatcct ggccggtttg tctgatgcca agctggcggc ctggccggcc agcttggccg
4561 ctgaagaaac cgagcgccgc cgtctaaaaa ggtgatgtgt atttgagtaa aacagcttgc
4621 gtcatgcggt cgctgcgtat atgatgcgat gagtaaataa acaaatacgc aaggggaacg
4681 catgaaggtt atcgctgtac ttaaccagaa aggcgggtca ggcaagacga ccatcgcaac
4741 ccatctagcc cgcgccctgc aactcgccgg ggccgatgtt ctgttagtcg attccgatcc
4801 ccagggcagt gcccgcgatt gggcggccgt gcgggaagat caaccgctaa ccgttgtcgg
4861 catcgaccgc ccgacgattg accgcgacgt gaaggccatc ggccggcgcg acttcgtagt
4921 gatcgacgga gcgccccagg cggcggactt ggctgtgtcc gcgatcaagg cagccgactt
4981 cgtgctgatt ccggtgcagc caagccctta cgacatatgg gcaaccgccg acctggtgga
5041 gctggttaag cagcgcattg aggtcacgga tggaaggcta caagcggcct ttgtcgtgtc
5101 gcgggcgatc aaaggcacgc gcatcggcgg tgaggttgcc gaggcgctgg ccgggtacga
5161 gctgcccatt cttgagtccc gtatcacgca gcgcgtgagc tacccaggca ctgccgccgc
5221 cggcacaacc gttcttgaat cagaacccga gggcgacgct gcccgcgagg tccaggcgct
5281 ggccgctgaa attaaatcaa aactcatttg agttaatgag gtaaagagaa aatgagcaaa
5341 agcacaaaca cgctaagtgc cggccgtccg agcgcacgca gcagcaaggc tgcaacgttg
5401 gccagcctgg cagacacgcc agccatgaag cgggtcaact ttcagttgcc ggcggaggat
5461 cacaccaagc tgaagatgta cgcggtacgc caaggcaaga ccattaccga gctgctatct
5521 gaatacatcg cgcagctacc agagtaaatg agcaaatgaa taaatgagta gatgaatttt
5581 agcggctaaa ggaggcggca tggaaaatca agaacaacca ggcaccgacg ccgtggaatg
5641 ccccatgtgt ggaggaacgg gcggttggcc aggcgtaagc ggctgggttg tctgccggcc
5701 ctgcaatggc actggaaccc ccaagcccga ggaatcggcg tgacggtcgc aaaccatccg
5761 gcccggtaca aatcggcgcg gcgctgggtg atgacctggt ggagaagttg aaggccgcgc
5821 aggccgccca gcggcaacgc atcgaggcag aagcacgccc cggtgaatcg tggcaagcgg
5881 ccgctgatcg aatccgcaaa gaatcccggc aaccgccggc agccggtgcg ccgtcgatta
5941 ggaagccgcc caagggcgac gagcaaccag attttttcgt tccgatgctc tatgacgtgg
6001 gcacccgcga tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg aagcgtgacc
6061 gacgagctgg cgaggtgatc cgctacgagc ttccagacgg gcacgtagag gtttccgcag
6121 ggccggccgg catggccagt gtgtgggatt acgacctggt actgatggcg gtttcccatc
6181 taaccgaatc catgaaccga taccgggaag ggaagggaga caagcccggc cgcgtgttcc
6241 gtccacacgt tgcggacgta ctcaagttct gccggcgagc cgatggcgga aagcagaaag
6301 acgacctggt agaaacctgc attcggttaa acaccacgca cgttgccatg cagcgtacga
6361 agaaggccaa gaacggccgc ctggtgacgg tatccgaggg tgaagccttg attagccgct
6421 acaagatcgt aaagagcgaa accgggcggc cggagtacat cgagatcgag ctagctgatt
6481 ggatgtaccg cgagatcaca gaaggcaaga acccggacgt gctgacggtt caccccgatt
6541 actttttgat cgatcccggc atcggccgtt ttctctaccg cctggcacgc cgcgccgcag
6601 gcaaggcaga agccagatgg ttgttcaaga cgatctacga acgcagtggc agcgccggag
6661 agttcaagaa gttctgtttc accgtgcgca agctgatcgg gtcaaatgac ctgccggagt
6721 acgatttgaa ggaggaggcg gggcaggctg gcccgatcct agtcatgcgc taccgcaacc
6781 tgatcgaggg cgaagcatcc gccggttcct aatgtacgga gcagatgcta gggcaaattg
6841 ccctagcagg ggaaaaaggt cgaaaaggtc tctttcctgt ggatagcacg tacattggga
6901 acccaaagcc gtacattggg aaccggaacc cgtacattgg gaacccaaag ccgtacattg
6961 ggaaccggtc acacatgtaa gtgactgata taaaagagaa aaaaggcgat ttttccgcct
7021 aaaactcttt aaaacttatt aaaactctta aaacccgcct ggcctgtgca taactgtctg
7081 gccagcgcac agccgaagag ctgcaaaaag cgcctaccct tcggtcgctg cgctccctac
7141 gccccgccgc ttcgcgtcgg cctatcgcgg ccgctggccg ctcaaaaatg gctggcctac
7201 ggccaggcaa tctaccaggg cgcggacaag ccgcgccgtc gccactcgac cgccggcgcc
7261 cacatcaagg caccctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
7321 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
7381 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
7441 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
7501 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
7561 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
7621 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
7681 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
7741 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
7801 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
7861 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
7921 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
7981 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
8041 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
8101 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
8161 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
8221 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
8281 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
8341 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
8401 tcatgcattc taggtactaa aacaattcat ccagtaaaat ataatatttt attttctccc
8461 aatcaggctt gatccccagt aagtcaaaaa atagctcgac atactgttct tccccgatat
8521 cctccctgat cgaccggacg cagaaggcaa tgtcatacca cttgtccgcc ctgccgcttc
8581 tcccaagatc aataaagcca cttactttgc catctttcac aaagatgttg ctgtctccca
8641 ggtcgccgtg ggaaaagaca agttcctctt cgggcttttc cgtctttaaa aaatcataca
8701 gctcgcgcgg atctttaaat ggagtgtctt cttcccagtt ttcgcaatcc acatcggcca
8761 gatcgttatt cagtaagtaa tccaattcgg ctaagcggct gtctaagcta ttcgtatagg
8821 gacaatccga tatgtcgatg gagtgaaaga gcctgatgca ctccgcatac agctcgataa
8881 tcttttcagg gctttgttca tcttcatact cttccgagca aaggacgcca tcggcctcac
8941 tcatgagcag attgctccag ccatcatgcc gttcaaagtg caggaccttt ggaacaggca
9001 gctttccttc cagccatagc atcatgtcct tttcccgttc aacatcatag gtggtccctt
9061 tataccggct gtccgtcatt tttaaatata ggttttcatt ttctcccacc agcttatata
9121 ccttagcagg agacattcct tccgtatctt ttacgcagcg gtatttttcg atcagttttt
9181 tcaattccgg tgatattctc attttagcca tttattattt ccttcctctt ttctacagta
9241 tttaaagata ccccaagaag ctaattataa caagacgaac tccaattcac tgttccttgc
9301 attctaaaac cttaaatacc agaaaacagc tttttcaaag ttgttttcaa agttggcgta
9361 taacatagta tcgacggagc cgattttgaa accgcggtga tcacaggcag caacgctctg
9421 tcatcgttac aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc
9481 ttagttgccg ttcttccgaa tagcatcggt aacatgagca aagtctgccg ccttacaacg
9541 gctctcccgc tgacgccgtc ccggactgat gggctgcctg tatcgagtgg tgattttgtg
9601 ccgagctgcc ggtcggggag ctgttggctg gctggtggca ggatatattg tggtgtaaac
9661 aaattgacgc ttagacaact taataacaca ttgcggacgt ttttaatgta ctgaattaac
9721 gccgaattaa ttcgggggat ctggatttta gtactggatt ttggttttag gaattagaaa
9781 ttttattgat agaagtattt tacaaataca aatacatact aagggtttct tatatgctca
9841 acacatgagc gaaaccctat aggaacccta attcccttat ctgggaacta ctcacacatt
9901 attatggaga aactcgagct tgtcgatcga cagatccggt cggcatctac tctatttctt
9961 tgccctcgga cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc
10021 atcggtccag acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc
10081 ggatcggacg attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc
10141 aaccaagctc tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagtcgtgg
10201 cgatcctgca agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc
10261 caaccacggc ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc
10321 ctcgctccag tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc
10381 gaaatccgcg tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc
10441 atcgagagcc tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata
10501 cacatgggga tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc
10561 ttgcggtccg aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc
10621 catagcctcc gcgaccggtt gtagaacagc gggcagttcg gtttcaggca ggtcttgcaa
10681 cgtgacaccc tgtgcacggc gggagatgca ataggtcagg ctctcgctaa actccccaat
10741 gtcaagcact tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc
10801 tttgtagaaa ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc
10861 gaagctgaaa gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc
10921 gaacttttcg atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttcatatc
10981 tcattgcccc ccggatctgc gaaagctcga gagagataga tttgtagaga gagactggtg
11041 atttcagcgt gtcctctcca aatgaaatga acttccttat atagaggaag gtcttgcgaa
11101 ggatagtggg attgtgcgtc atcccttacg tcagtggaga tatcacatca atccacttgc
11161 tttgaagacg tggttggaac gtcttctttt tccacgatgc tcctcgtggg tgggggtcca
11221 tctttgggac cactgtcggc agaggcatct tgaacgatag cctttccttt atcgcaatga
11281 tggcatttgt aggtgccacc ttccttttct actgtccttt tgatgaagtg acagatagct
11341 gggcaatgga atccgaggag gtttcccgat attacccttt gttgaaaagt ctcaatagcc
11401 ctttggtctt ctgagactgt atctttgata ttcttggagt agacgagagt gtcgtgctcc
11461 accatgttat cacatcaatc cacttgcttt gaagacgtgg ttggaacgtc ttctttttcc
11521 acgatgctcc tcgtgggtgg gggtccatct ttgggaccac tgtcggcaga ggcatcttga
11581 acgatagcct ttcctttatc gcaatgatgg catttgtagg tgccaccttc cttttctact
11641 gtccttttga tgaagtgaca gatagctggg caatggaatc cgaggaggtt tcccgatatt
11701 accctttgtt gaaaagtctc aatagccctt tggtcttctg agactgtatc tttgatattc
11761 ttggagtaga cgagagtgtc gtgctccacc atgttggcaa gctgctctag ccaatacgca
11821 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
11881 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
11941 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
12001 aatttcacac aggaaacagc tatgaccatg attac
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
| P3195/pCMV-SPORT6-ASNSD1（1点突变1同义突变）人源基因质粒 |
| P3196/pCMV-SPORT6-ARFGAP3（1点突变2同义突变）人源基因质粒 |
| P3197/pCMV-SPORT6-POLG2（506-1458bp）人源基因质粒 |
| P3198/pCMV-SPORT6-SRP54（1同义突变）人源基因质粒 |
| P3199/pCMV-SPORT6-ELMO2（1同义突变）人源基因质粒 |
| P3200/pCMV-SPORT6-SLC12A9（1同义突变）人源基因质粒 |
| P3201/pCMV-SPORT6-CARHSP1（3同义突变）人源基因质粒 |
| P3202/pCMV-SPORT6-SOX15人源基因质粒 |
| P3204/pCMV-SPORT6-XPO5人源基因质粒 |
| P3205/pCMV-SPORT6-C1orf43人源基因质粒 |
| P3206/pCMV-SPORT6-HPS1（1同义突变）人源基因质粒 |
| P3207/pCMV-SPORT6-METTL5人源基因质粒 |
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ws 各位站友，本人因克隆某个植物抗病基因而完成了其BAC文库，目前正在做图位克隆的工作。 但文库的完成耗费了大量人力、物力和财力，因此想请教下该库还有什么其它用途。或与其 它技术结合，可开展什么工作。 谢谢了。。。。 zhujoker BAC (Bacterial Artificial Chromosome, 细菌人工染色体)文库是进行全基因组测序、构建物理图谱、染色体步
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pCXUN-FLAG植物表达质粒
￥100 - 1000
