| 质粒名称: | pTetQCas-BsaI |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 170629 |
| 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: | 7955 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 |
| 插入大小: | 951 bp |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | pTetQCas-BsaI was a gift from Sheng Yang (Addgene plasmid # 170629 ; http://n2t.net/addgene:170629 ; RRID:Addgene_170629) |
| 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
pTetQCas-BsaI载体质粒基本信息
pTetQCas-BsaI质粒图谱载体图谱和pTetQCas-BsaI载体序列质粒序列多克隆位点信息
pTetQCas-BsaI质粒载体简介
pTetQCas-BsaI质粒序列载体序列
LOCUS pTetQCas-BsaI 7955 bp ds-DNA circular SYN 03-AUG-2021
DEFINITION Expression of CRISPR-associated transposases.
ACCESSION .
VERSION .
KEYWORDS pTetQCas-BsaI
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7955)
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 7955)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 3, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7955
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 19..74
/gene="tetR"
/label=tetR/tetA promoters
/note="overlapping promoters for bacterial tetR and tetA"
protein_bind 25..43
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
protein_bind 55..73
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
promoter 281..336
/gene="tetR"
/label=tetR/tetA promoters
/note="overlapping promoters for bacterial tetR and tetA"
protein_bind 287..305
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
protein_bind 317..335
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind complement(5140..5158)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 5154..5201
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(5372..6163)
/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(6164..6255)
/gene="bla"
/label=AmpR promoter
CDS complement(7332..7955)
/codon_start=1
/gene="tetR from transposon Tn10"
/product="tetracycline repressor TetR"
/label=TetR
/note="TetR binds to the tetracycline operator tetO to
inhibit transcription. This inhibition can be relieved by
adding tetracycline or doxycycline."
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
primer_bind 7863..7882
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
ORIGIN
1 cattaattcc taatttttgt tgacactcta tcattgatag agttatttta ccactcccta
61 tcagtgatag agaaaagtga actctagaaa taattttgtt taactttaaa aggagatata
121 ccatgggtga actgccgagt aggtagctga taacgagacc tctggtctcg tgaactgccg
181 agtaggtagc tgataacgga tccgaattcg agctcggcgc gcctgcaggt cgacaagctt
241 gcggccgctc taatctagac atcattaatt cctaattttt gttgacactc tatcattgat
301 agagttattt taccactccc tatcagtgat agagaaaagt gaactctaga aataattttg
361 tttaacttta aaggagatat acatatgttt ttgcaaagac ctaaacctta cagcgatgaa
421 agtttagaaa gtttctttat ccgagtggct aacaaaaatg gctacggtga tgtccatcgc
481 ttcctagaag ccactaaacg attccttcaa gacattgacc ataatggcta tcaaaccttt
541 ccgactgata taactcggat aaacccatac tcagctaaaa acagttccag cgcacgaact
601 gcgtcattcc tgaaacttgc acaattgaca tttaatgaac cgccagagct acttgggttg
661 gcaattaaca gaacaaacat gaaatactcg ccgtcaacta gcgcggttgt tagaggtgca
721 gaagtctttc ctcgcagttt actacggacg cactccatcc cctgctgtcc tttgtgtctg
781 cgagaaaatg gctacgcctc ctacctttgg cactttcagg ggtacgaata ctgccacagc
841 cataacgtac ctttaattac cacttgtagc tgtggtaagg agtttgacta ccgagtatct
901 gggttaaagg gcatttgctg caaatgcaag gagcctatca ccttaaccag cagggagaac
961 ggtcatgagg cagcgtgtac tgtttcaaac tggcttgctg gccatgaatc taaacctctg
1021 ccaaatcttc ctaaaagcta ccgatggggt ttagttcatt ggtggatggg tattaaagat
1081 agcgagttcg atcacttttc gttcgttcaa tttttctcaa actggccaag gtcattccac
1141 tcgataatcg aagatgaagt agagttcaac cttgagcatg ctgttgtcag cacgtctgaa
1201 ttacgactaa aagatcttct tggtcgattg tttttcggtt caattcggtt acctgagcgg
1261 aatcttcaac acaatatcat ccttggtgag cttctctgct atttagaaaa tcgtttatgg
1321 caagacaagg gattaatcgc caacctcaaa atgaacgcgt tagaggcgac tgtaatgtta
1381 aattgtagcc tcgatcagat tgcatcaatg gttgaacaac gcatcttgaa gccaaatcga
1441 aaaagcaagc ccaacagccc tcttgatgtt accgattatc tatttcattt cggcgatatt
1501 ttctgtcttt ggttagctga gttccaaagc gatgagttta accgttcgtt ttatgtgtcg
1561 aggtggtaaa tgcaaactct gaaagaacta atcgcatcca atcctgacga cttaacaact
1621 gaacttaaga gagcatttcg tccactcaca ccgcatattg caattgatgg taatgaactt
1681 gacgcactga cgatattagt caatttaacc gataagactg atgatcagaa agacctgctc
1741 gatcgagcca aatgcaagca aaaacttcga gatgaaaaat ggtgggctag ctgcataaat
1801 tgcgttaatt acagacaaag ccataaccca aaattcccgg atatacgttc tgaaggcgtg
1861 attcgaaccc aagccctggg tgaattaccg agtttcctac tctcgtcctc aaaaatccca
1921 ccataccatt ggtcatatag ccatgattca aaatacgtca acaaaagcgc attcctcacc
1981 aatgagtttt gttgggatgg tgagatctca tgtttgggtg agcttcttaa agatgcagat
2041 cacccacttt ggaatacttt gaaaaagtta ggttgttctc aaaaaacttg caaagcaatg
2101 gcaaaacaac tagctgatat tactctcacg actatcaatg tcacactcgc accaaattac
2161 ctgactcaaa tatctctccc cgatagtgat acatcctaca tttcactctc tcctgttgca
2221 tcgctatcga tgcaaagcca ctttcatcag aggctccaag atgaaaatcg gcatagcgcg
2281 ataacgcggt ttagccgaac taccaacatg ggggtcactg cgatgacatg tggcggcgca
2341 tttagaatgt taaagtctgg cgctaagttt tccagccccc ctcatcaccg attaaacagt
2401 aaacgaagtt ggttgacgtc agagcatgtt cagtcattaa aacagtacca gcgcctcaac
2461 aaaagcctca tacctgaaaa ctctcggatt gcactccgta gaaaatacaa aatcgagctt
2521 caaaatatgg tcagatcttg gtttgcaatg caagaccata cccttgattc gaacatactt
2581 atccaacatt tgaatcatga cctatcttac ttaggagcca caaaacgttt tgcatacgat
2641 ccagcgatga ccaagctctt tactgagctt ttgaaacgag agttatcaaa ttcaatcaat
2701 aatggtgagc aacacactaa tggatcgttt ttagtcctac cgaatatcag ggtttgtggc
2761 gcaacagctt taagctcccc ggtaacggtg gggattccat cacttacagc tttctttggc
2821 ttcgttcacg catttgaacg gaatataaat cgcaccacct catcgtttcg tgttgaatcc
2881 tttgcgatat gcgtccatca actacatgtc gagaagcgag gtttgacagc agagtttgtg
2941 gaaaaaggcg acgggactat atccgctccc gcgacccggg atgactggca gtgtgatgtc
3001 gtatttagcc ttattttgaa caccaacttt gctcaacata ttgaccaaga tacgttagtt
3061 acatcactac caaagcgatt ggctcggggt tcagcaaaaa ttgcgattga tgactttaaa
3121 catatcaact cattctcgac attagaaaca gcgatcgaat ctctgccaat agaagctggt
3181 aggtggttat cactttacgc acagtcaaac aataatctaa gtgatctatt agcagccatg
3241 acagaggacc atcagctcat ggcaagctgc gtcggttacc acttgttaga agagcccaaa
3301 gataaaccaa actccctcag aggttacaaa cacgctatcg ccgagtgcat cattggactc
3361 attaactcaa tcacctttag ctcagagact gatcccaaca caatcttttg gtcgctaaag
3421 aactatcaaa actacctagt ggtacagcca aggagtatca acgatgaaac taccgacaaa
3481 tctagcctat gagcgctcta tcgacccatc agatgtctgt ttttttgtcg tctggcccga
3541 tgatagaaaa acacctttaa cctacaattc tcgtactctg ctcgggcaaa tggaagcggc
3601 atcattagcc tatgatgtct caggtcaacc aataaaaagt gccaccgctg aggcgttagc
3661 tcaagggaac cctcatcaag ttgatttctg ccacgttcca tacggtgcga gtcatattga
3721 atgcagtttc tccgtctcgt tttcttctga actacgtcaa ccatataagt gtaactcaag
3781 caaagttaaa caaacgctag tgcaattagt cgagctctac gaaacgaaaa tcggctggac
3841 tgagctagca acccgatatt tgatgaacat ttgcaacggt aaatggctgt ggaaaaatac
3901 ccgtaaagcc tattgctgga acattgtact tacaccttgg ccttggaacg gggaaaaggt
3961 tggatttgaa gatattcgta ctaactacac ctcacggcaa gactttaaaa ataataaaaa
4021 ttggtctgct atagttgaaa tgatcaaaac cgcattttct agtactgatg ggctggcgat
4081 atttgaagtc agggccacct tgcacttgcc aacgaatgct atggtgcggc caagccaagt
4141 tttcacagaa aaagaaagtg gcagtaaaag taaatctaaa actcaaaaca gtcgagtttt
4201 tcagagtaca actattgatg gtgaacgatc gccaatacta ggggccttta aaacgggagc
4261 agctattgca accattgacg actggtatcc tgaagccact gagccactaa gggtcggacg
4321 gtttggggtt catcgcgaag atgtcacttg ctaccgtcat ccgtctaccg gaaaagattt
4381 tttctcgata ttacaacaag cagagcacta tattgaagtg ttgagcgcca acaaaactcc
4441 cgctcaagaa actatcaacg acatgcactt tttaatggct aacctgatta agggtgggat
4501 gttccagcat aaaggagact gactgtgaaa tggtattata agacaatcac ctttctgcca
4561 gagttgtgca acaacgagtc actggctgca aagtgtctcc gcgttctgca tggatttaac
4621 tatcagtatg agacacgaaa tatcggcgtt tcatttccgc tttggtgtga tgcaacggtt
4681 ggaaaaaaga tttcatttgt cagcaagaac aagatagaac tcgacttact acttaaacaa
4741 cactatttcg tccaaatgga acaacttcaa tactttcata tatccaacac tgttctcgtc
4801 ccagaagatt gtacatacgt ttcctttaga cgctgtcaat ctatagataa gctcacagca
4861 gcagggctgg caaggaaaat cagacgcctg gagaaacgtg ctctatctag aggcgagcaa
4921 tttgacccat catcttttgc tcaaaaagag catactgcaa tagcgcacta ccactcactt
4981 ggggagtcca gcaaacagac gaaccgcaac tttcgactca atatcaggat gctctcggag
5041 caaccccgtg aggggaactc gatttttagt agctatggct tatcaaattc agaaaactcg
5101 tttcagcctg tacccttaat ctgacctagg ctgctgccac cgctgagcaa taactagcat
5161 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaacct caggcatttg
5221 agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag caatagacat
5281 aagcggctat ttaacgaccc tgccctgaac cgacgaccgg gtcatcgtgg ccggatcttg
5341 cggcccctcg gcttgaacga attgttagac attatttgcc gactaccttg gtgatctcgc
5401 ctttcacgta gtggacaaat tcttccaact gatctgcgcg cgaggccaag cgatcttctt
5461 cttgtccaag ataagcctgt ctagcttcaa gtatgacggg ctgatactgg gccggcaggc
5521 gctccattgc ccagtcggca gcgacatcct tcggcgcgat tttgccggtt actgcgctgt
5581 accaaatgcg ggacaacgta agcactacat ttcgctcatc gccagcccag tcgggcggcg
5641 agttccatag cgttaaggtt tcatttagcg cctcaaatag atcctgttca ggaaccggat
5701 caaagagttc ctccgccgct ggacctacca aggcaacgct atgttctctt gcttttgtca
5761 gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat
5821 tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg ataacgccac ggaatgatgt
5881 cgtcgtgcac aacaatggtg acttctacag cgcggagaat ctcgctctct ccaggggaag
5941 ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg
6001 tcaccgtaac cagcaaatca atatcactgt gtggcttcag gccgccatcc actgcggagc
6061 cgtacaaatg tacggccagc aacgtcggtt cgagatggcg ctcgatgacg ccaactacct
6121 ctgatagttg agtcgatact tcggcgatca ccgcttccct catactcttc ctttttcaat
6181 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
6241 agaaaaataa acaaatagct agctcactcg gtcgctacgc tccgggcgtg agactgcggc
6301 gggcgctgcg gacacataca aagttaccca cagattccgt ggataagcag gggactaaca
6361 tgtgaggcaa aacagcaggg ccgcgccggt ggcgtttttc cataggctcc gccctcctgc
6421 cagagttcac ataaacagac gcttttccgg tgcatctgtg ggagccgtga ggctcaacca
6481 tgaatctgac agtacgggcg aaacccgaca ggacttaaag atccccaccg tttccggcgg
6541 gtcgctccct cttgcgctct cctgttccga ccctgccgtt taccggatac ctgttccgcc
6601 tttctccctt acgggaagtg tggcgctttc tcatagctca cacactggta tctcggctcg
6661 gtgtaggtcg ttcgctccaa gctgggctgt aagcaagaac tccccgttca gcccgactgc
6721 tgcgccttat ccggtaactg ttcacttgag tccaacccgg aaaagcacgg taaaacgcca
6781 ctggcagcag ccattggtaa ctgggagttc gcagaggatt tgtttagcta aacacgcggt
6841 tgctcttgaa gtgtgcgcca aagtccggct acactggaag gacagatttg gttgctgtgc
6901 tctgcgaaag ccagttacca cggttaagca gttccccaac tgacttaacc ttcgatcaaa
6961 ccacctcccc aggtggtttt ttcgaaggtt aagtcagttg gggaactgct taaccttcga
7021 tcaaaccacc tccccaggtg gttttttcgt ttacagggca aaagattacg cgcagaaaaa
7081 aaggatctca agaagatcct ttgatctttt ctactgaacc gctctagatt tcagtgcaat
7141 ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat
7201 gttagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
7261 cctgatgaat cccctaatga tttttatcaa aatcattaag gttaccatca cggaaaaagg
7321 ttatgctgct tttaagaccc actttcacat ttaagttgtt tttctaatcc gcatatgatc
7381 aattcaaggc cgaataagaa ggctggctct gcaccttggt gatcaaataa ttcgatagct
7441 tgtcgtaata atggcggcat actatcagta gtaggtgttt ccctttcttc tttagcgact
7501 tgatgctctt gatcttccaa tacgcaacct aaagtaaaat gccccacagc gctgagtgca
7561 tataatgcat tctctagtga aaaaccttgt tggcataaaa aggctaattg attttcgaga
7621 gtttcatact gtttttctgt aggccgtgta cctaaatgta cttttgctcc atcgcgatga
7681 cttagtaaag cacatctaaa acttttagcg ttattacgta aaaaatcttg ccagctttcc
7741 ccttctaaag ggcaaaagtg agtatggtgc ctatctaaca tctcaatggc taaggcgtcg
7801 agcaaagccc gcttattttt tacatgccaa tacaatgtag gctgctctac acctagcttc
7861 tgggcgagtt tacgggttgt taaaccttcg attccgacct cattaagcag ctctaatgcg
7921 ctgttaatca ctttactttt atctaatcta gacat
//
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