| 质粒名称: | pAraQCas-BsaI |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 170633 |
| 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: | 8674 bp |
| 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: | pAraQCas-BsaI was a gift from Sheng Yang (Addgene plasmid # 170633 ; http://n2t.net/addgene:170633 ; RRID:Addgene_170633) |
| 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 pAraQCas-BsaI 8674 bp ds-DNA circular SYN 03-AUG-2021
DEFINITION Expression of CRISPR-associated transposases.
ACCESSION .
VERSION .
KEYWORDS pAraQCas-BsaI
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8674)
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 8674)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 3, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8674
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(67..85)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 81..128
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(299..1090)
/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(1091..1182)
/gene="bla"
/label=AmpR promoter
CDS complement(2318..3196)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
promoter 3223..3507
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
promoter 3778..3943
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
ORIGIN
1 tatggcttat caaattcaga aaactcgttt cagcctgtac ccttaatctg acctaggctg
61 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
121 gttttttgct gaaacctcag gcatttgaga agcacacggt cacactgctt ccggtagtca
181 ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc cctgaaccga
241 cgaccgggtc atcgtggccg gatcttgcgg cccctcggct tgaacgaatt gttagacatt
301 atttgccgac taccttggtg atctcgcctt tcacgtagtg gacaaattct tccaactgat
361 ctgcgcgcga ggccaagcga tcttcttctt gtccaagata agcctgtcta gcttcaagta
421 tgacgggctg atactgggcc ggcaggcgct ccattgccca gtcggcagcg acatccttcg
481 gcgcgatttt gccggttact gcgctgtacc aaatgcggga caacgtaagc actacatttc
541 gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt taaggtttca tttagcgcct
601 caaatagatc ctgttcagga accggatcaa agagttcctc cgccgctgga cctaccaagg
661 caacgctatg ttctcttgct tttgtcagca agatagccag atcaatgtcg atcgtggctg
721 gctcgaagat acctgcaaga atgtcattgc gctgccattc tccaaattgc agttcgcgct
781 tagctggata acgccacgga atgatgtcgt cgtgcacaac aatggtgact tctacagcgc
841 ggagaatctc gctctctcca ggggaagccg aagtttccaa aaggtcgttg atcaaagctc
901 gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag caaatcaata tcactgtgtg
961 gcttcaggcc gccatccact gcggagccgt acaaatgtac ggccagcaac gtcggttcga
1021 gatggcgctc gatgacgcca actacctctg atagttgagt cgatacttcg gcgatcaccg
1081 cttccctcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
1141 tgagcggata catatttgaa tgtatttaga aaaataaaca aatagctagc tcactcggtc
1201 gctacgctcc gggcgtgaga ctgcggcggg cgctgcggac acatacaaag ttacccacag
1261 attccgtgga taagcagggg actaacatgt gaggcaaaac agcagggccg cgccggtggc
1321 gtttttccat aggctccgcc ctcctgccag agttcacata aacagacgct tttccggtgc
1381 atctgtggga gccgtgaggc tcaaccatga atctgacagt acgggcgaaa cccgacagga
1441 cttaaagatc cccaccgttt ccggcgggtc gctccctctt gcgctctcct gttccgaccc
1501 tgccgtttac cggatacctg ttccgccttt ctcccttacg ggaagtgtgg cgctttctca
1561 tagctcacac actggtatct cggctcggtg taggtcgttc gctccaagct gggctgtaag
1621 caagaactcc ccgttcagcc cgactgctgc gccttatccg gtaactgttc acttgagtcc
1681 aacccggaaa agcacggtaa aacgccactg gcagcagcca ttggtaactg ggagttcgca
1741 gaggatttgt ttagctaaac acgcggttgc tcttgaagtg tgcgccaaag tccggctaca
1801 ctggaaggac agatttggtt gctgtgctct gcgaaagcca gttaccacgg ttaagcagtt
1861 ccccaactga cttaaccttc gatcaaacca cctccccagg tggttttttc gaaggttaag
1921 tcagttgggg aactgcttaa ccttcgatca aaccacctcc ccaggtggtt ttttcgttta
1981 cagggcaaaa gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
2041 ctgaaccgct ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc
2101 ccatacgata taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct
2161 gactgggttg aaggctctca agggcatcag tatattttag atgaagatta tttcttaatc
2221 tagacatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
2281 gcacatttcc ccgaaaagtg ccacctgcat cgatttatta tgacaacttg acggctacat
2341 cattcacttt ttcttcacaa ccggcacgga actcgctcgg gctggccccg gtgcattttt
2401 taaatacccg cgagaagtag agttgatcgt caaaaccaac attgcgaccg acggtggcga
2461 taggcatccg ggtggtgctc aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc
2521 agcttaagac gctaatccct aactgctggc ggaaaagatg tgacagacgc gacggcgaca
2581 agcaaacatg ctgtgcgacg ctggcgatat caaaattgct gtctgccagg tgatcgctga
2641 tgtactgaca agcctcgcgt acccgattat ccatcggtgg atggagcgac tcgttaatcg
2701 cttccatgcg ccgcagtaac aattgctcaa gcagatttat cgccagcagc tccgaatagc
2761 gcccttcccc ttgcccggcg ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt
2821 gcgcttcatc cgggcgaaag aaccccgtat tggcaaatat tgacggccag ttaagccatt
2881 catgccagta ggcgcgcgga cgaaagtaaa cccactggtg ataccattcg cgagcctccg
2941 gatgacgacc gtagtgatga atctctcctg gcgggaacag caaaatatca cccggtcggc
3001 aaacaaattc tcgtccctga tttttcacca ccccctgacc gcgaatggtg agattgagaa
3061 tataaccttt cattcccagc ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa
3121 tcggcgttaa acccgccacc agatgggcat taaacgagta tcccggcagc aggggatcat
3181 tttgcgcttc agccatactt ttcatactcc cgccattcag agaagaaacc aattgtccat
3241 attgcatcag acattgccgt cactgcgtct tttactggct cttctcgcta accaaaccgg
3301 taaccccgct tattaaaagc attctgtaac aaagcgggac caaagccatg acaaaaacgc
3361 gtaacaaaag tgtctataat cacggcagaa aagtccacat tgattatttg cacggcgtca
3421 cactttgcta tgccatagca tttttatcca taagattagc ggatcctacc tgacgctttt
3481 tatcgcaact ctctactgtt tctccatacc cgtttttttg ggaattcgag ctctaaagga
3541 gatataccat gggtgaactg ccgagtaggt agctgataac gagacctctg gtctcgtgaa
3601 ctgccgagta ggtagctgat aacggatccg aattcgagct cggcgcgcct gcaggtcgac
3661 aagcttgcgg ccgcatattg catcagacat tgccgtcact gcgtctttta ctggctcttc
3721 tcgctaacca aaccggtaac cccgcttatt aaaagcattc tgtaacaaag cgggaccaaa
3781 gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg gcagaaaagt ccacattgat
3841 tatttgcacg gcgtcacact ttgctatgcc atagcatttt tatccataag attagcggat
3901 cctacctgac gctttttatc gcaactctct actgtttctc catacccgtt tttttgggaa
3961 ttcgagctct aaaggagata tacatatgtt tttgcaaaga cctaaacctt acagcgatga
4021 aagtttagaa agtttcttta tccgagtggc taacaaaaat ggctacggtg atgtccatcg
4081 cttcctagaa gccactaaac gattccttca agacattgac cataatggct atcaaacctt
4141 tccgactgat ataactcgga taaacccata ctcagctaaa aacagttcca gcgcacgaac
4201 tgcgtcattc ctgaaacttg cacaattgac atttaatgaa ccgccagagc tacttgggtt
4261 ggcaattaac agaacaaaca tgaaatactc gccgtcaact agcgcggttg ttagaggtgc
4321 agaagtcttt cctcgcagtt tactacggac gcactccatc ccctgctgtc ctttgtgtct
4381 gcgagaaaat ggctacgcct cctacctttg gcactttcag gggtacgaat actgccacag
4441 ccataacgta cctttaatta ccacttgtag ctgtggtaag gagtttgact accgagtatc
4501 tgggttaaag ggcatttgct gcaaatgcaa ggagcctatc accttaacca gcagggagaa
4561 cggtcatgag gcagcgtgta ctgtttcaaa ctggcttgct ggccatgaat ctaaacctct
4621 gccaaatctt cctaaaagct accgatgggg tttagttcat tggtggatgg gtattaaaga
4681 tagcgagttc gatcactttt cgttcgttca atttttctca aactggccaa ggtcattcca
4741 ctcgataatc gaagatgaag tagagttcaa ccttgagcat gctgttgtca gcacgtctga
4801 attacgacta aaagatcttc ttggtcgatt gtttttcggt tcaattcggt tacctgagcg
4861 gaatcttcaa cacaatatca tccttggtga gcttctctgc tatttagaaa atcgtttatg
4921 gcaagacaag ggattaatcg ccaacctcaa aatgaacgcg ttagaggcga ctgtaatgtt
4981 aaattgtagc ctcgatcaga ttgcatcaat ggttgaacaa cgcatcttga agccaaatcg
5041 aaaaagcaag cccaacagcc ctcttgatgt taccgattat ctatttcatt tcggcgatat
5101 tttctgtctt tggttagctg agttccaaag cgatgagttt aaccgttcgt tttatgtgtc
5161 gaggtggtaa atgcaaactc tgaaagaact aatcgcatcc aatcctgacg acttaacaac
5221 tgaacttaag agagcatttc gtccactcac accgcatatt gcaattgatg gtaatgaact
5281 tgacgcactg acgatattag tcaatttaac cgataagact gatgatcaga aagacctgct
5341 cgatcgagcc aaatgcaagc aaaaacttcg agatgaaaaa tggtgggcta gctgcataaa
5401 ttgcgttaat tacagacaaa gccataaccc aaaattcccg gatatacgtt ctgaaggcgt
5461 gattcgaacc caagccctgg gtgaattacc gagtttccta ctctcgtcct caaaaatccc
5521 accataccat tggtcatata gccatgattc aaaatacgtc aacaaaagcg cattcctcac
5581 caatgagttt tgttgggatg gtgagatctc atgtttgggt gagcttctta aagatgcaga
5641 tcacccactt tggaatactt tgaaaaagtt aggttgttct caaaaaactt gcaaagcaat
5701 ggcaaaacaa ctagctgata ttactctcac gactatcaat gtcacactcg caccaaatta
5761 cctgactcaa atatctctcc ccgatagtga tacatcctac atttcactct ctcctgttgc
5821 atcgctatcg atgcaaagcc actttcatca gaggctccaa gatgaaaatc ggcatagcgc
5881 gataacgcgg tttagccgaa ctaccaacat gggggtcact gcgatgacat gtggcggcgc
5941 atttagaatg ttaaagtctg gcgctaagtt ttccagcccc cctcatcacc gattaaacag
6001 taaacgaagt tggttgacgt cagagcatgt tcagtcatta aaacagtacc agcgcctcaa
6061 caaaagcctc atacctgaaa actctcggat tgcactccgt agaaaataca aaatcgagct
6121 tcaaaatatg gtcagatctt ggtttgcaat gcaagaccat acccttgatt cgaacatact
6181 tatccaacat ttgaatcatg acctatctta cttaggagcc acaaaacgtt ttgcatacga
6241 tccagcgatg accaagctct ttactgagct tttgaaacga gagttatcaa attcaatcaa
6301 taatggtgag caacacacta atggatcgtt tttagtccta ccgaatatca gggtttgtgg
6361 cgcaacagct ttaagctccc cggtaacggt ggggattcca tcacttacag ctttctttgg
6421 cttcgttcac gcatttgaac ggaatataaa tcgcaccacc tcatcgtttc gtgttgaatc
6481 ctttgcgata tgcgtccatc aactacatgt cgagaagcga ggtttgacag cagagtttgt
6541 ggaaaaaggc gacgggacta tatccgctcc cgcgacccgg gatgactggc agtgtgatgt
6601 cgtatttagc cttattttga acaccaactt tgctcaacat attgaccaag atacgttagt
6661 tacatcacta ccaaagcgat tggctcgggg ttcagcaaaa attgcgattg atgactttaa
6721 acatatcaac tcattctcga cattagaaac agcgatcgaa tctctgccaa tagaagctgg
6781 taggtggtta tcactttacg cacagtcaaa caataatcta agtgatctat tagcagccat
6841 gacagaggac catcagctca tggcaagctg cgtcggttac cacttgttag aagagcccaa
6901 agataaacca aactccctca gaggttacaa acacgctatc gccgagtgca tcattggact
6961 cattaactca atcaccttta gctcagagac tgatcccaac acaatctttt ggtcgctaaa
7021 gaactatcaa aactacctag tggtacagcc aaggagtatc aacgatgaaa ctaccgacaa
7081 atctagccta tgagcgctct atcgacccat cagatgtctg tttttttgtc gtctggcccg
7141 atgatagaaa aacaccttta acctacaatt ctcgtactct gctcgggcaa atggaagcgg
7201 catcattagc ctatgatgtc tcaggtcaac caataaaaag tgccaccgct gaggcgttag
7261 ctcaagggaa ccctcatcaa gttgatttct gccacgttcc atacggtgcg agtcatattg
7321 aatgcagttt ctccgtctcg ttttcttctg aactacgtca accatataag tgtaactcaa
7381 gcaaagttaa acaaacgcta gtgcaattag tcgagctcta cgaaacgaaa atcggctgga
7441 ctgagctagc aacccgatat ttgatgaaca tttgcaacgg taaatggctg tggaaaaata
7501 cccgtaaagc ctattgctgg aacattgtac ttacaccttg gccttggaac ggggaaaagg
7561 ttggatttga agatattcgt actaactaca cctcacggca agactttaaa aataataaaa
7621 attggtctgc tatagttgaa atgatcaaaa ccgcattttc tagtactgat gggctggcga
7681 tatttgaagt cagggccacc ttgcacttgc caacgaatgc tatggtgcgg ccaagccaag
7741 ttttcacaga aaaagaaagt ggcagtaaaa gtaaatctaa aactcaaaac agtcgagttt
7801 ttcagagtac aactattgat ggtgaacgat cgccaatact aggggccttt aaaacgggag
7861 cagctattgc aaccattgac gactggtatc ctgaagccac tgagccacta agggtcggac
7921 ggtttggggt tcatcgcgaa gatgtcactt gctaccgtca tccgtctacc ggaaaagatt
7981 ttttctcgat attacaacaa gcagagcact atattgaagt gttgagcgcc aacaaaactc
8041 ccgctcaaga aactatcaac gacatgcact ttttaatggc taacctgatt aagggtggga
8101 tgttccagca taaaggagac tgactgtgaa atggtattat aagacaatca cctttctgcc
8161 agagttgtgc aacaacgagt cactggctgc aaagtgtctc cgcgttctgc atggatttaa
8221 ctatcagtat gagacacgaa atatcggcgt ttcatttccg ctttggtgtg atgcaacggt
8281 tggaaaaaag atttcatttg tcagcaagaa caagatagaa ctcgacttac tacttaaaca
8341 acactatttc gtccaaatgg aacaacttca atactttcat atatccaaca ctgttctcgt
8401 cccagaagat tgtacatacg tttcctttag acgctgtcaa tctatagata agctcacagc
8461 agcagggctg gcaaggaaaa tcagacgcct ggagaaacgt gctctatcta gaggcgagca
8521 atttgaccca tcatcttttg ctcaaaaaga gcatactgca atagcgcact accactcact
8581 tggggagtcc agcaaacaga cgaaccgcaa ctttcgactc aatatcagga tgctctcgga
8641 gcaaccccgt gaggggaact cgatttttag tagc
//
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