| 质粒名称: | pUC18-mini-Tn7T-Plac-dCas9 (Plac) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 105235 |
| 存储实验室: | Kristala Prather |
|
载体骨架基本信息 |
|
| 载体名称: | pUC18 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5981 bp |
| 完整质粒大小: | 9404 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达,CRISPR,合成生物学 |
| 筛选标记: | 庆大霉素 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | S. pasteurianus dCas9 |
| 物种来源: | S. pasteurianus |
| 插入大小: | 3423 bp |
| 基因突变: | D10A, H599A |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | HA(插入件上的N端子) |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pUC18-mini-Tn7T-Plac-dCas9 (Plac) was a gift from Kristala Prather (Addgene plasmid # 105235 ; http://n2t.net/addgene:105235 ; RRID:Addgene_105235) |
| 参考文献部分: | A Robust CRISPR Interference Gene Repression System in Pseudomonas. Tan SZ, Reisch CR, Prather KLJ. J Bacteriol. 2018 Mar 12;200(7). pii: JB.00575-17. doi: 10.1128/JB.00575-17. Print 2018 Apr 1. 10.1128/JB.00575-17 PubMed 29311279 |
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pUC18-mini-Tn7T-Plac-dCas9 (Plac)载体质粒基本信息
pUC18-mini-Tn7T-Plac-dCas9 (Plac)质粒图谱载体图谱和pUC18-mini-Tn7T-Plac-dCas9 (Plac)载体序列质粒序列多克隆位点信息
质粒图谱.png)
pUC18-mini-Tn7T-Plac-dCas9 (Plac)质粒载体简介
pUC18-mini-Tn7T-Plac-dCas9 (Plac)质粒序列载体序列
LOCUS pUC18-mini-Tn7T-Plac-dCas9 (Plac) 9404 bp ds-DNA circular SYN 07-AUG-2018
DEFINITION Tn7 integrating plasmid with S. pasteurianus dCas9 expressed from
the Plac promoter for expression in P. aeruginosa.
ACCESSION .
VERSION .
KEYWORDS pUC18-mini-Tn7T-Plac-dCas9 (Plac)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9404)
AUTHORS Tan SZ, Reisch CR, Prather KLJ
TITLE A Robust CRISPR Interference Gene Repression System in Pseudomonas.
JOURNAL J Bacteriol. 2018 Mar 12;200(7). pii: JB.00575-17. doi:
10.1128/JB.00575-17. Print 2018 Apr 1.
PUBMED 29311279
REFERENCE 2 (bases 1 to 9404)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 7, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9404
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 100..119
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 533..549
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 534..550
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
terminator 578..672
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
terminator 775..861
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
protein_bind 907..954
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS complement(1084..1617)
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(1806..1834)
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
protein_bind 1875..1922
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS complement(1965..3047)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=lacI
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 3009..3028
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(3048..3125)
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
promoter 3355..3385
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3393..3409
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 3398..3420
/label=M13/pUC Reverse
/note="In lacZ gene"
CDS 3461..3487
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
mobile_element 7099..7264
/mobile_element_type="transposon:Tn7"
/note="mini-Tn7 element (left end of the Tn7 transposon)"
rep_origin complement(7534..8122)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7614..7633)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(8293..9153)
/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"
primer_bind 8916..8935
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9154..9258)
/gene="bla"
/label=AmpR promoter
primer_bind 9326..9344
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(9382..9404)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
ORIGIN
1 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
61 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
121 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
181 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
241 cttcgctatt acgccagcta gaggaccagc cgcgtaacct ggcaaaatcg gttacggttg
301 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa
361 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga
421 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact
481 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggg gttcgaggtc
541 gacggtatcg ataagctagc ttaattagct gagcttggac tcctgttgat agatccagta
601 atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt
661 attggtgaga atccaagcta gactgcgatg agtggcaggg cggggcgtaa tttttttaag
721 gcagttattg gtgcccttaa acgcctgggg taatgactct ctagcttgag gcatcaaata
781 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
841 gctctcctga gtaggacaaa tccgccgcta ggagcttgcg gcccggacga tcgaattggg
901 gatcttgaag tacctattcc gaagttccta ttctctagaa agtataggaa cttcagagcg
961 cttttgaagc tgatgtgctt aaaaacttac tcaatggaat tctagatcga attggccgcg
1021 gcgttgtgac aatttaccga acaactccgc ggccgggaag ccgatctcgg cttgaacgaa
1081 ttgttaggtg gcggtacttg ggtcgatatc aaagtgcatc acttcttccc gtatgcccaa
1141 ctttgtatag agagccactg cgggatcgtc accgtaatct gcttgcacgt agatcacata
1201 agcaccaagc gcgttggcct catgcttgag gagattgatg agcgcggtgg caatgccctg
1261 cctccggtgc tcgccggaga ctgcgagatc atagatatag atctcactac gcggctgctc
1321 aaacttgggc agaacgtaag ccgcgagagc gccaacaacc gcttcttggt cgaaggcagc
1381 aagcgcgatg aatgtcttac tacggagcaa gttcccgagg taatcggagt ccggctgatg
1441 ttgggagtag gtggctacgt ctccgaactc acgaccgaaa agatcaagag cagcccgcat
1501 ggatttgact tggtcagggc cgagcctaca tgtgcgaatg atgcccatac ttgagccacc
1561 taactttgtt ttagggcgac tgccctgctg cgtaacatcg ttgctgctgc gtaacatcgt
1621 tgctgctcca taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc
1681 gaggcataga ctgtacaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt
1741 taccaccgct gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc
1801 attacagttt acgaaccgaa caggcttatg tcaattcgat ccattgctgt tgacaaaggg
1861 aatcagggga tcttgaagtt cctattccga agttcctatt ctctagaaag tataggaact
1921 tcagagcgct tttgaagcta attcgatcat gcattgcgtt gcgctcactg cccgctttcc
1981 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
2041 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
2101 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
2161 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
2221 gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg
2281 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt aggaacgatg
2341 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
2401 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
2461 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
2521 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
2581 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
2641 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
2701 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
2761 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
2821 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
2881 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
2941 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
3001 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
3061 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgca ccattcgatg
3121 gtgtcaacgt aaatgccgct tcgccttcgc gcgcgaattg caagctgatc cgggcttatc
3181 gactgcacgg tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct
3241 gtgcaggtcg taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata
3301 atgttttttg cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgtttaca
3361 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
3421 aaacagaatt cgagctcctg actcaggagg tatcgatatg tacccatacg atgttccaga
3481 ttacgctatg accaacggca agatcctggg cctggccatc ggcatcgcct cggtgggcgt
3541 gggcatcatc gaggccaaga ccggcaaggt ggtgcacgcc aactcgcgcc tgttctcggc
3601 cgccaacgcc gagaacaacg ccgagcgccg cggcttccgc ggctcgcgcc gcctgaaccg
3661 ccgcaagaag caccgcgtga agcgcgtgcg cgacctgttc gagaagtacg gcatcgtgac
3721 cgacttccgc aacctgaacc tgaacccgta cgagctgcgc gtgaagggcc tgaccgagca
3781 gctgaagaac gaggagctgt tcgccgccct gcgcaccatc tcgaagcgcc gcggcatctc
3841 gtacctggac gacgccgagg acgactcgac cggctcgacc gactacgcca agtcgatcga
3901 cgagaaccgc cgcctgctga agaacaagac cccgggccag atccagctgg agcgcctgga
3961 gaagtacggc cagctgcgcg gcaacttcac cgtgtacgac gagaacggcg aggcccaccg
4021 cctgatcaac gtgttctcga cctcggacta cgagaaggag gcccgcaaga tcctggagac
4081 ccaggccgac tacaacaaga agatcaccgc cgagttcatc gacgactacg tggagatcct
4141 gacccagaag cgcaagtact accacggccc gggcaacgag aagtcgcgca ccgactacgg
4201 ccgcttccgc accgacggca ccaccctgga gaacatcttc ggcatcctga tcggcaagtg
4261 caacttctac ccggacgagt accgcgcctc gaaggcctcg tacaccgccc aggagtacaa
4321 cttcctgaac gacctgaaca acctgaaggt gtcgaccgag accggcaagc tgtcgaccga
4381 gcagaaggag tcgctggtgg agttcgccaa gaacaccgcc accctgggcc cggccaagct
4441 gctgaaggag atcgccaaga tcctggactg caaggtggac gagatcaagg gctaccgcga
4501 ggacgacaag ggcaagccgg acctgcacac cttcgagccg taccgcaagc tgaagttcaa
4561 cctggagtcg atcaacatcg acgacctgtc gcgcgaggtg atcgacaagc tggccgacat
4621 cctgaccctg aacaccgagc gcgagggcat cgaggacgcc atcaagcgca acctgccgaa
4681 ccagttcacc gaggagcaga tctcggagat catcaaggtg cgcaagtcgc agtcgaccgc
4741 cttcaacaag ggctggcact cgttctcggc caagctgatg aacgagctga tcccggagct
4801 gtacgccacc tcggacgagc agatgaccat cctgacccgc ctggagaagt tcaaggtgaa
4861 caagaagtcg tcgaagaaca ccaagaccat cgacgagaag gaggtgaccg acgagatcta
4921 caacccggtg gtggccaagt cggtgcgcca gaccatcaag atcatcaacg ccgccgtgaa
4981 gaagtacggc gacttcgaca agatcgtgat cgagatgccg cgcgacaaga acgccgacga
5041 cgagaagaag ttcatcgaca agcgcaacaa ggagaacaag aaggagaagg acgacgccct
5101 gaagcgcgcc gcctacctgt acaactcgtc ggacaagctg ccggacgagg tgttccacgg
5161 caacaagcag ctggagacca agatccgcct gtggtaccag cagggcgagc gctgcctgta
5221 ctcgggcaag ccgatctcga tccaggagct ggtgcacaac tcgaacaact tcgagatcga
5281 cgccatcctg ccgctgtcgc tgtcgttcga cgactcgctg gccaacaagg tgctggtgta
5341 cgcctggacc aaccaggaga agggccaaaa gaccccgtac caggtgatcg actcgatgga
5401 cgccgcctgg tcgttccgcg agatgaagga ctacgtgctg aagcagaagg gcctgggcaa
5461 gaagaagcgc gactacctgc tgaccaccga gaacatcgac aagatcgagg tgaagaagaa
5521 gttcatcgag cgcaacctgg tggacacccg ctacgcctcg cgcgtggtgc tgaactcgct
5581 gcagtcggcc ctgcgcgagc tgggcaagga caccaaggtg tcggtggtgc gcggccagtt
5641 cacctcgcag ctgcgccgca agtggaagat cgacaagtcg cgcgagacct accaccacca
5701 cgccgtggac gccctgatca tcgccgcctc gtcgcagctg aagctgtggg agaagcagga
5761 caacccgatg ttcgtggact acggcaagaa ccaggtggtg gacaagcaga ccggcgagat
5821 cctgtcggtg tcggacgacg agtacaagga gctggtgttc cagccgccgt accagggctt
5881 cgtgaacacc atctcgtcga agggcttcga ggacgagatc ctgttctcgt accaggtgga
5941 ctcgaagtac aaccgcaagg tgtcggacgc caccatctac tcgacccgca aggccaagat
6001 cggcaaggac aagaaggagg agacctacgt gctgggcaag atcaaggaca tctactcgca
6061 gaacggcttc gacaccttca tcaagaagta caacaaggac aagacccagt tcctgatgta
6121 ccagaaggac tcgctgacct gggagaacgt gatcgaggtg atcctgcgcg actacccgac
6181 caccaagaag tcggaggacg gcaagaacga cgtgaagtgc aacccgttcg aggagtaccg
6241 ccgcgagaac ggcctgatct gcaagtactc gaagaagggc aagggcaccc cgatcaagtc
6301 gctgaagtac tacgacaaga agctgggcaa ctgcatcgac atcaccccgg aggagtcgcg
6361 caacaaggtg atcctgcagt cgatcaaccc gtggcgcgcc gacgtgtact tcaacccgga
6421 gaccctgaag tacgagctga tgggcctgaa gtactcggac ctgtcgttcg agaagggcac
6481 cggcaactac cacatctcgc aggagaagta cgacgccatc aaggagaagg agggcatcgg
6541 caagaagtcg gagttcaagt tcaccctgta ccgcaacgac ctgatcctga tcaaggacat
6601 cgcctcgggc gagcaggaga tctaccgctt cctgtcgcgc accatgccga acgtgaacca
6661 ctacgtggag ctgaagccgt acgacaagga gaagttcgac aacgtgcagg agctggtgga
6721 ggccctgggc gaggccgaca aggtgggccg ctgcatcaag ggcctgaaca agccgaacat
6781 ctcgatctac aaggtgcgca ccgacgtgct gggcaacaag tacttcgtga agaagaaggg
6841 cgacaagccg aagctggact tcaagaacaa caagaagtaa atactctggc catttttctt
6901 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcaacgcgtg
6961 cggccgctga ttagctaagc agaaggccat cctgacggat gcctttttcg cttgctagct
7021 gcagcccggg ggatcccccg ggctgcagga attcctcgag aagcttgggc ccggtacctc
7081 gcgaaggcct tgcaggccaa ccagataagt gaaatctagt tccaaactat tttgtcattt
7141 ttaattttcg tattagctta cgacgctaca cccagttccc atctattttg tcactcttcc
7201 ctaaataatc cttaaaaact ccatttccac ccctcccagt tcccaactat tttgtccgcc
7261 cacagcgggg catttttctt cctgttatgt ttgggcgctg cattaatgaa tcggccaacg
7321 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgacccgct
7381 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
7441 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
7501 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
7561 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
7621 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
7681 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
7741 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
7801 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
7861 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
7921 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
7981 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
8041 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
8101 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
8161 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
8221 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
8281 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
8341 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
8401 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
8461 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
8521 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
8581 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
8641 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
8701 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
8761 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
8821 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
8881 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
8941 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
9001 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
9061 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
9121 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
9181 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
9241 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
9301 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
9361 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccgg
//
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pUC18-mini-Tn7T-Plac-dCas9 (Plac)质粒载体应用举例
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