| 质粒名称: | pRhaQCas-BsaI |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 170631 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pCDFDuet-1 |
| 完整质粒大小: | 9450bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | Streptomycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | VchTniQ, VchCas8, VchCas7, VchCas6, CRISPR(BsaI) |
| 物种来源: | Vibrio cholera |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | pRhaQCas-BsaI was a gift from Sheng Yang (Addgene plasmid # 170631 ; http://n2t.net/addgene:170631 ; RRID:Addgene_170631) |
| 参考文献部分: | Programming Cells by Multicopy Chromosomal Integration Using CRISPR-Associated Transposases. Zhang Y, Yang J, Yang S, Zhang J, Chen J, Tao R, Jiang Y, Yang J, Yang S. CRISPR J. 2021 Jun;4(3):350-359. doi: 10.1089/crispr.2021.0018. 10.1089/crispr.2021.0018 PubMed 34152213 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pRhaQCas-BsaI 9450 bp ds-DNA circular SYN 10-NOV-2021
DEFINITION Expression of CRISPR-associated transposases.
ACCESSION .
VERSION .
KEYWORDS pRhaQCas-BsaI
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9450)
AUTHORS Zhang Y, Yang J, Yang S, Zhang J, Chen J, Tao R, Jiang Y, Yang J,
Yang S
TITLE Programming Cells by Multicopy Chromosomal Integration Using
CRISPR-Associated Transposases.
JOURNAL CRISPR J. 2021 Jun;4(3):350-359. doi: 10.1089/crispr.2021.0018.
PUBMED 34152213
REFERENCE 2 (bases 1 to 9450)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 10, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9450
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(135..983)
/codon_start=1
/gene="E. coli rhaR"
/product="transcriptional activator of rhaR and rhaS"
/label=rhaR
/note="needed for rhamnose-dependent activation of the rhaB
promoter"
/translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
D"
CDS complement(1057..1893)
/codon_start=1
/gene="E. coli rhaS"
/product="positive regulator of the rhaB promoter
"
/label=rhaS
/note="activated by L-rhamnose"
/translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
promoter 2039..2157
/label=rhaB promoter
/note="promoter of the E. coli rhaBAD operon, conferring
tight induction with L-rhamnose and repression with
D-glucose in the presence of RhaR and RhaS (Giacalone et
al., 2006)"
promoter 2453..2571
/label=rhaB promoter
/note="promoter of the E. coli rhaBAD operon, conferring
tight induction with L-rhamnose and repression with
D-glucose in the presence of RhaR and RhaS (Giacalone et
al., 2006)"
primer_bind complement(7344..7362)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 7358..7405
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(7576..8367)
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
promoter complement(8368..8459)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ggctctcaag ggcatcggtt cttatggctc ttgtatctat cagtgaagca tcaagactaa
61 caaacaaaag tagaacaact gttcaccgtt acatatcaaa gggaaaactg tccataccca
121 tggggcatgg cgaattaatc tttctgcgaa ttgagatgac gccactggct gggcgtcatc
181 ccggtttccc gggtaaacac caccgaaaaa tagttactat cttcaaagcc acattcggtc
241 gaaatatcac tgattaacag gcggctatgc tggagaagat attgcgcatg acacactctg
301 acctgtcgca gatattgatt gatggtcatt ccagtctgct ggcgaaattg ctgacgcaaa
361 acgcgctcac tgcacgatgc ctcatcacaa aatttatcca gcgcaaaggg acttttcagg
421 ctagccgcca gccgggtaat cagcttatcc agcaacgttt cgctggatgt tggcggcaac
481 gaatcactgg tgtaacgatg gcgattcagc aacatcacca actgcccgaa cagcaactca
541 gccatttcgt tagcaaacgg cacatgctga ctactttcat gctcaagctg accgataacc
601 tgccgcgcct gcgccatccc catgctacct aagcgccagt gtggttgccc tgcgctggcg
661 ttaaatcccg gaatcgcccc ctgccagtca agattcagct tcagacgctc cgggcaataa
721 ataatattct gcaaaaccag atcgttaacg gaagcgtagg agtgtttatc gtcagcatga
781 atgtaaaaga gatcgccacg ggtaatgcga taagggcgat cgttgagtac atgcaggcca
841 ttaccgcgcc agacaatcac cagctcacaa aaatcatgtg tatgttcagc aaagacatct
901 tgcggataac ggtcagccac agcgactgcc tgctggtcgc tggcaaaaaa atcatctttg
961 agaagtttta actgatgcgc caccgtggct acctcggcca gagaacgaag ttgattattc
1021 gcaatatggc gtacaaatac gttgagaaga ttcgcgttat tgcagaaagc catcccgtcc
1081 ctggcgaata tcacgcggtg accagttaaa ctctcggcga aaaagcgtcg aaaagtggtt
1141 actgtcgctg aatccacagc gataggcgat gtcagtaacg ctggcctcgc tgtggcgtag
1201 cagatgtcgg gctttcatca gtcgcaggcg gttcaggtat cgctgaggcg tcagtcccgt
1261 ttgctgctta agctgccgat gtagcgtacg cagtgaaaga gaaaattgat ccgccacggc
1321 atcccaattc acctcatcgg caaaatggtc ctccagccag gccagaagca agttgagacg
1381 tgatgcgctg ttttccaggt tctcctgcaa actgctttta cgcagcaaga gcagtaattg
1441 cataaacaag atctcgcgac tggcggtcga gggtaaatca ttttcccctt cctgctgttc
1501 catctgtgca accagctgtc gcacctgctg caatacgctg tggttaacgc gccagtgaga
1561 cggatactgc ccatccagct cttgtggcag caactgattc agcccggcga gaaactgaaa
1621 tcgatccggc gagcgataca gcacattggt cagacacaga ttatcggtat gttcatacag
1681 atgccgatca tgatcgcgta cgaaacagac cgtgccaccg gtgatggtat agggctgccc
1741 attaaacaca tgaatacccg tgccatgttc gacaatcaca atttcatgaa aatcatgatg
1801 atgttcagga aaatccgcct gcgggagccg gggttctatc gccacggacg cgttaccaga
1861 cggaaaaaaa tccacactat gtaatacggt catactggcc tcctgatgtc gtcaacacgg
1921 cgaaatagta atcacgaggt caggttctta ccttaaattt tcgacggaaa accacgtaaa
1981 aaacgtcgat ttttcaagat acagcgtgaa ttttcaggaa atgcggtgag catcacatca
2041 ccacaattca gcaaattgtg aacatcatca cgttcatctt tccctggttg ccaatggccc
2101 attttcctgt cagtaacgag aaggtcgcga attcaggcgc tttttagact ggtcgtaaag
2161 gagatatacc atgggtgaac tgccgagtag gtagctgata acgagacctc tggtctcgtg
2221 aactgccgag taggtagctg ataacggatc cgaattcgag ctcggcgcgc ctgcaggtcg
2281 acaagcttgc ggccgcaata cggtcatact ggcctcctga tgtcgtcaac acggcgaaat
2341 agtaatcacg aggtcaggtt cttaccttaa attttcgacg gaaaaccacg taaaaaacgt
2401 cgatttttca agatacagcg tgaattttca ggaaatgcgg tgagcatcac atcaccacaa
2461 ttcagcaaat tgtgaacatc atcacgttca tctttccctg gttgccaatg gcccattttc
2521 ctgtcagtaa cgagaaggtc gcgaattcag gcgcttttta gactggtcgt aataaaggag
2581 atatacatat gtttttgcaa agacctaaac cttacagcga tgaaagttta gaaagtttct
2641 ttatccgagt ggctaacaaa aatggctacg gtgatgtcca tcgcttccta gaagccacta
2701 aacgattcct tcaagacatt gaccataatg gctatcaaac ctttccgact gatataactc
2761 ggataaaccc atactcagct aaaaacagtt ccagcgcacg aactgcgtca ttcctgaaac
2821 ttgcacaatt gacatttaat gaaccgccag agctacttgg gttggcaatt aacagaacaa
2881 acatgaaata ctcgccgtca actagcgcgg ttgttagagg tgcagaagtc tttcctcgca
2941 gtttactacg gacgcactcc atcccctgct gtcctttgtg tctgcgagaa aatggctacg
3001 cctcctacct ttggcacttt caggggtacg aatactgcca cagccataac gtacctttaa
3061 ttaccacttg tagctgtggt aaggagtttg actaccgagt atctgggtta aagggcattt
3121 gctgcaaatg caaggagcct atcaccttaa ccagcaggga gaacggtcat gaggcagcgt
3181 gtactgtttc aaactggctt gctggccatg aatctaaacc tctgccaaat cttcctaaaa
3241 gctaccgatg gggtttagtt cattggtgga tgggtattaa agatagcgag ttcgatcact
3301 tttcgttcgt tcaatttttc tcaaactggc caaggtcatt ccactcgata atcgaagatg
3361 aagtagagtt caaccttgag catgctgttg tcagcacgtc tgaattacga ctaaaagatc
3421 ttcttggtcg attgtttttc ggttcaattc ggttacctga gcggaatctt caacacaata
3481 tcatccttgg tgagcttctc tgctatttag aaaatcgttt atggcaagac aagggattaa
3541 tcgccaacct caaaatgaac gcgttagagg cgactgtaat gttaaattgt agcctcgatc
3601 agattgcatc aatggttgaa caacgcatct tgaagccaaa tcgaaaaagc aagcccaaca
3661 gccctcttga tgttaccgat tatctatttc atttcggcga tattttctgt ctttggttag
3721 ctgagttcca aagcgatgag tttaaccgtt cgttttatgt gtcgaggtgg taaatgcaaa
3781 ctctgaaaga actaatcgca tccaatcctg acgacttaac aactgaactt aagagagcat
3841 ttcgtccact cacaccgcat attgcaattg atggtaatga acttgacgca ctgacgatat
3901 tagtcaattt aaccgataag actgatgatc agaaagacct gctcgatcga gccaaatgca
3961 agcaaaaact tcgagatgaa aaatggtggg ctagctgcat aaattgcgtt aattacagac
4021 aaagccataa cccaaaattc ccggatatac gttctgaagg cgtgattcga acccaagccc
4081 tgggtgaatt accgagtttc ctactctcgt cctcaaaaat cccaccatac cattggtcat
4141 atagccatga ttcaaaatac gtcaacaaaa gcgcattcct caccaatgag ttttgttggg
4201 atggtgagat ctcatgtttg ggtgagcttc ttaaagatgc agatcaccca ctttggaata
4261 ctttgaaaaa gttaggttgt tctcaaaaaa cttgcaaagc aatggcaaaa caactagctg
4321 atattactct cacgactatc aatgtcacac tcgcaccaaa ttacctgact caaatatctc
4381 tccccgatag tgatacatcc tacatttcac tctctcctgt tgcatcgcta tcgatgcaaa
4441 gccactttca tcagaggctc caagatgaaa atcggcatag cgcgataacg cggtttagcc
4501 gaactaccaa catgggggtc actgcgatga catgtggcgg cgcatttaga atgttaaagt
4561 ctggcgctaa gttttccagc ccccctcatc accgattaaa cagtaaacga agttggttga
4621 cgtcagagca tgttcagtca ttaaaacagt accagcgcct caacaaaagc ctcatacctg
4681 aaaactctcg gattgcactc cgtagaaaat acaaaatcga gcttcaaaat atggtcagat
4741 cttggtttgc aatgcaagac catacccttg attcgaacat acttatccaa catttgaatc
4801 atgacctatc ttacttagga gccacaaaac gttttgcata cgatccagcg atgaccaagc
4861 tctttactga gcttttgaaa cgagagttat caaattcaat caataatggt gagcaacaca
4921 ctaatggatc gtttttagtc ctaccgaata tcagggtttg tggcgcaaca gctttaagct
4981 ccccggtaac ggtggggatt ccatcactta cagctttctt tggcttcgtt cacgcatttg
5041 aacggaatat aaatcgcacc acctcatcgt ttcgtgttga atcctttgcg atatgcgtcc
5101 atcaactaca tgtcgagaag cgaggtttga cagcagagtt tgtggaaaaa ggcgacggga
5161 ctatatccgc tcccgcgacc cgggatgact ggcagtgtga tgtcgtattt agccttattt
5221 tgaacaccaa ctttgctcaa catattgacc aagatacgtt agttacatca ctaccaaagc
5281 gattggctcg gggttcagca aaaattgcga ttgatgactt taaacatatc aactcattct
5341 cgacattaga aacagcgatc gaatctctgc caatagaagc tggtaggtgg ttatcacttt
5401 acgcacagtc aaacaataat ctaagtgatc tattagcagc catgacagag gaccatcagc
5461 tcatggcaag ctgcgtcggt taccacttgt tagaagagcc caaagataaa ccaaactccc
5521 tcagaggtta caaacacgct atcgccgagt gcatcattgg actcattaac tcaatcacct
5581 ttagctcaga gactgatccc aacacaatct tttggtcgct aaagaactat caaaactacc
5641 tagtggtaca gccaaggagt atcaacgatg aaactaccga caaatctagc ctatgagcgc
5701 tctatcgacc catcagatgt ctgttttttt gtcgtctggc ccgatgatag aaaaacacct
5761 ttaacctaca attctcgtac tctgctcggg caaatggaag cggcatcatt agcctatgat
5821 gtctcaggtc aaccaataaa aagtgccacc gctgaggcgt tagctcaagg gaaccctcat
5881 caagttgatt tctgccacgt tccatacggt gcgagtcata ttgaatgcag tttctccgtc
5941 tcgttttctt ctgaactacg tcaaccatat aagtgtaact caagcaaagt taaacaaacg
6001 ctagtgcaat tagtcgagct ctacgaaacg aaaatcggct ggactgagct agcaacccga
6061 tatttgatga acatttgcaa cggtaaatgg ctgtggaaaa atacccgtaa agcctattgc
6121 tggaacattg tacttacacc ttggccttgg aacggggaaa aggttggatt tgaagatatt
6181 cgtactaact acacctcacg gcaagacttt aaaaataata aaaattggtc tgctatagtt
6241 gaaatgatca aaaccgcatt ttctagtact gatgggctgg cgatatttga agtcagggcc
6301 accttgcact tgccaacgaa tgctatggtg cggccaagcc aagttttcac agaaaaagaa
6361 agtggcagta aaagtaaatc taaaactcaa aacagtcgag tttttcagag tacaactatt
6421 gatggtgaac gatcgccaat actaggggcc tttaaaacgg gagcagctat tgcaaccatt
6481 gacgactggt atcctgaagc cactgagcca ctaagggtcg gacggtttgg ggttcatcgc
6541 gaagatgtca cttgctaccg tcatccgtct accggaaaag attttttctc gatattacaa
6601 caagcagagc actatattga agtgttgagc gccaacaaaa ctcccgctca agaaactatc
6661 aacgacatgc actttttaat ggctaacctg attaagggtg ggatgttcca gcataaagga
6721 gactgactgt gaaatggtat tataagacaa tcacctttct gccagagttg tgcaacaacg
6781 agtcactggc tgcaaagtgt ctccgcgttc tgcatggatt taactatcag tatgagacac
6841 gaaatatcgg cgtttcattt ccgctttggt gtgatgcaac ggttggaaaa aagatttcat
6901 ttgtcagcaa gaacaagata gaactcgact tactacttaa acaacactat ttcgtccaaa
6961 tggaacaact tcaatacttt catatatcca acactgttct cgtcccagaa gattgtacat
7021 acgtttcctt tagacgctgt caatctatag ataagctcac agcagcaggg ctggcaagga
7081 aaatcagacg cctggagaaa cgtgctctat ctagaggcga gcaatttgac ccatcatctt
7141 ttgctcaaaa agagcatact gcaatagcgc actaccactc acttggggag tccagcaaac
7201 agacgaaccg caactttcga ctcaatatca ggatgctctc ggagcaaccc cgtgagggga
7261 actcgatttt tagtagctat ggcttatcaa attcagaaaa ctcgtttcag cctgtaccct
7321 taatctgacc taggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
7381 taaacgggtc ttgaggggtt ttttgctgaa acctcaggca tttgagaagc acacggtcac
7441 actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg ctatttaacg
7501 accctgccct gaaccgacga ccgggtcatc gtggccggat cttgcggccc ctcggcttga
7561 acgaattgtt agacattatt tgccgactac cttggtgatc tcgcctttca cgtagtggac
7621 aaattcttcc aactgatctg cgcgcgaggc caagcgatct tcttcttgtc caagataagc
7681 ctgtctagct tcaagtatga cgggctgata ctgggccggc aggcgctcca ttgcccagtc
7741 ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa
7801 cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc atagcgttaa
7861 ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga gttcctccgc
7921 cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga tagccagatc
7981 aatgtcgatc gtggctggct cgaagatacc tgcaagaatg tcattgcgct gccattctcc
8041 aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt gcacaacaat
8101 ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag tttccaaaag
8161 gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg taaccagcaa
8221 atcaatatca ctgtgtggct tcaggccgcc atccactgcg gagccgtaca aatgtacggc
8281 cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata gttgagtcga
8341 tacttcggcg atcaccgctt ccctcatact cttccttttt caatattatt gaagcattta
8401 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
8461 agctagctca ctcggtcgct acgctccggg cgtgagactg cggcgggcgc tgcggacaca
8521 tacaaagtta cccacagatt ccgtggataa gcaggggact aacatgtgag gcaaaacagc
8581 agggccgcgc cggtggcgtt tttccatagg ctccgccctc ctgccagagt tcacataaac
8641 agacgctttt ccggtgcatc tgtgggagcc gtgaggctca accatgaatc tgacagtacg
8701 ggcgaaaccc gacaggactt aaagatcccc accgtttccg gcgggtcgct ccctcttgcg
8761 ctctcctgtt ccgaccctgc cgtttaccgg atacctgttc cgcctttctc ccttacggga
8821 agtgtggcgc tttctcatag ctcacacact ggtatctcgg ctcggtgtag gtcgttcgct
8881 ccaagctggg ctgtaagcaa gaactccccg ttcagcccga ctgctgcgcc ttatccggta
8941 actgttcact tgagtccaac ccggaaaagc acggtaaaac gccactggca gcagccattg
9001 gtaactggga gttcgcagag gatttgttta gctaaacacg cggttgctct tgaagtgtgc
9061 gccaaagtcc ggctacactg gaaggacaga tttggttgct gtgctctgcg aaagccagtt
9121 accacggtta agcagttccc caactgactt aaccttcgat caaaccacct ccccaggtgg
9181 ttttttcgaa ggttaagtca gttggggaac tgcttaacct tcgatcaaac cacctcccca
9241 ggtggttttt tcgtttacag ggcaaaagat tacgcgcaga aaaaaaggat ctcaagaaga
9301 tcctttgatc ttttctactg aaccgctcta gatttcagtg caatttatct cttcaaatgt
9361 agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgttagt catgccccgc
9421 gcccaccgga aggagctgac tgggttggaa
//
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