| 质粒名称: | pTetQCas-8+IS186 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 170636 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pUC19 |
| 完整质粒大小: | 8478bp |
| 载体类型: | Bacterial Expression |
| 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(8+IS186 array) |
| 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: | pTetQCas-8+IS186 was a gift from Sheng Yang (Addgene plasmid # 170636 ; http://n2t.net/addgene:170636 ; RRID:Addgene_170636) |
| 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 pTetQCas-8+IS186 8478 bp ds-DNA circular SYN 15-NOV-2021
DEFINITION Expression of CRISPR-associated transposases.
ACCESSION .
VERSION .
KEYWORDS pTetQCas-8+IS186
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8478)
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 8478)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 15, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8478
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 497..552
/gene="tetR"
/label=tetR/tetA promoters
/note="overlapping promoters for bacterial tetR and tetA"
protein_bind 503..521
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
protein_bind 533..551
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind complement(5356..5374)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 5370..5417
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(5588..6379)
/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(6380..6471)
/gene="bla"
/label=AmpR promoter
CDS complement(7548..8171)
/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 8079..8098
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
promoter 8190..8245
/gene="tetR"
/label=tetR/tetA promoters
/note="overlapping promoters for bacterial tetR and tetA"
protein_bind 8196..8214
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
protein_bind 8226..8244
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
1 tgaactgccg agtaggtagc tgataacgtc agagtggcgt atccgatgaa tcaccacagg
61 tgaactgccg agtaggtagc tgataacgca gaagtgcacg gcgtgtgcgg cggcgagcag
121 tgaactgccg agtaggtagc tgataactcg atccgtgggt gtttatgttc tcagcaggag
181 tgaactgccg agtaggtagc tgataactta cttacggcgt tgcgcattct cattgcaccg
241 tgaactgccg agtaggtagc tgataacgca ttactaagct ggcgcgtgaa gctgtagaag
301 tgaactgccg agtaggtagc tgataacggg gcgtaaaaag aaggggcgct gattaacgcg
361 tgaactgccg agtaggtagc tgataacgtt taaactaggg ataacagggt aatggatccg
421 aattcgagct cggcgcgcct gcaggtcgac aagcttgcgg ccgctctaat ctagacatca
481 ttaattccta atttttgttg acactctatc attgatagag ttattttacc actccctatc
541 agtgatagag aaaagtgaac tctagaaata attttgttta actttaaagg agatatacat
601 atgtttttgc aaagacctaa accttacagc gatgaaagtt tagaaagttt ctttatccga
661 gtggctaaca aaaatggcta cggtgatgtc catcgcttcc tagaagccac taaacgattc
721 cttcaagaca ttgaccataa tggctatcaa acctttccga ctgatataac tcggataaac
781 ccatactcag ctaaaaacag ttccagcgca cgaactgcgt cattcctgaa acttgcacaa
841 ttgacattta atgaaccgcc agagctactt gggttggcaa ttaacagaac aaacatgaaa
901 tactcgccgt caactagcgc ggttgttaga ggtgcagaag tctttcctcg cagtttacta
961 cggacgcact ccatcccctg ctgtcctttg tgtctgcgag aaaatggcta cgcctcctac
1021 ctttggcact ttcaggggta cgaatactgc cacagccata acgtaccttt aattaccact
1081 tgtagctgtg gtaaggagtt tgactaccga gtatctgggt taaagggcat ttgctgcaaa
1141 tgcaaggagc ctatcacctt aaccagcagg gagaacggtc atgaggcagc gtgtactgtt
1201 tcaaactggc ttgctggcca tgaatctaaa cctctgccaa atcttcctaa aagctaccga
1261 tggggtttag ttcattggtg gatgggtatt aaagatagcg agttcgatca cttttcgttc
1321 gttcaatttt tctcaaactg gccaaggtca ttccactcga taatcgaaga tgaagtagag
1381 ttcaaccttg agcatgctgt tgtcagcacg tctgaattac gactaaaaga tcttcttggt
1441 cgattgtttt tcggttcaat tcggttacct gagcggaatc ttcaacacaa tatcatcctt
1501 ggtgagcttc tctgctattt agaaaatcgt ttatggcaag acaagggatt aatcgccaac
1561 ctcaaaatga acgcgttaga ggcgactgta atgttaaatt gtagcctcga tcagattgca
1621 tcaatggttg aacaacgcat cttgaagcca aatcgaaaaa gcaagcccaa cagccctctt
1681 gatgttaccg attatctatt tcatttcggc gatattttct gtctttggtt agctgagttc
1741 caaagcgatg agtttaaccg ttcgttttat gtgtcgaggt ggtaaatgca aactctgaaa
1801 gaactaatcg catccaatcc tgacgactta acaactgaac ttaagagagc atttcgtcca
1861 ctcacaccgc atattgcaat tgatggtaat gaacttgacg cactgacgat attagtcaat
1921 ttaaccgata agactgatga tcagaaagac ctgctcgatc gagccaaatg caagcaaaaa
1981 cttcgagatg aaaaatggtg ggctagctgc ataaattgcg ttaattacag acaaagccat
2041 aacccaaaat tcccggatat acgttctgaa ggcgtgattc gaacccaagc cctgggtgaa
2101 ttaccgagtt tcctactctc gtcctcaaaa atcccaccat accattggtc atatagccat
2161 gattcaaaat acgtcaacaa aagcgcattc ctcaccaatg agttttgttg ggatggtgag
2221 atctcatgtt tgggtgagct tcttaaagat gcagatcacc cactttggaa tactttgaaa
2281 aagttaggtt gttctcaaaa aacttgcaaa gcaatggcaa aacaactagc tgatattact
2341 ctcacgacta tcaatgtcac actcgcacca aattacctga ctcaaatatc tctccccgat
2401 agtgatacat cctacatttc actctctcct gttgcatcgc tatcgatgca aagccacttt
2461 catcagaggc tccaagatga aaatcggcat agcgcgataa cgcggtttag ccgaactacc
2521 aacatggggg tcactgcgat gacatgtggc ggcgcattta gaatgttaaa gtctggcgct
2581 aagttttcca gcccccctca tcaccgatta aacagtaaac gaagttggtt gacgtcagag
2641 catgttcagt cattaaaaca gtaccagcgc ctcaacaaaa gcctcatacc tgaaaactct
2701 cggattgcac tccgtagaaa atacaaaatc gagcttcaaa atatggtcag atcttggttt
2761 gcaatgcaag accataccct tgattcgaac atacttatcc aacatttgaa tcatgaccta
2821 tcttacttag gagccacaaa acgttttgca tacgatccag cgatgaccaa gctctttact
2881 gagcttttga aacgagagtt atcaaattca atcaataatg gtgagcaaca cactaatgga
2941 tcgtttttag tcctaccgaa tatcagggtt tgtggcgcaa cagctttaag ctccccggta
3001 acggtgggga ttccatcact tacagctttc tttggcttcg ttcacgcatt tgaacggaat
3061 ataaatcgca ccacctcatc gtttcgtgtt gaatcctttg cgatatgcgt ccatcaacta
3121 catgtcgaga agcgaggttt gacagcagag tttgtggaaa aaggcgacgg gactatatcc
3181 gctcccgcga cccgggatga ctggcagtgt gatgtcgtat ttagccttat tttgaacacc
3241 aactttgctc aacatattga ccaagatacg ttagttacat cactaccaaa gcgattggct
3301 cggggttcag caaaaattgc gattgatgac tttaaacata tcaactcatt ctcgacatta
3361 gaaacagcga tcgaatctct gccaatagaa gctggtaggt ggttatcact ttacgcacag
3421 tcaaacaata atctaagtga tctattagca gccatgacag aggaccatca gctcatggca
3481 agctgcgtcg gttaccactt gttagaagag cccaaagata aaccaaactc cctcagaggt
3541 tacaaacacg ctatcgccga gtgcatcatt ggactcatta actcaatcac ctttagctca
3601 gagactgatc ccaacacaat cttttggtcg ctaaagaact atcaaaacta cctagtggta
3661 cagccaagga gtatcaacga tgaaactacc gacaaatcta gcctatgagc gctctatcga
3721 cccatcagat gtctgttttt ttgtcgtctg gcccgatgat agaaaaacac ctttaaccta
3781 caattctcgt actctgctcg ggcaaatgga agcggcatca ttagcctatg atgtctcagg
3841 tcaaccaata aaaagtgcca ccgctgaggc gttagctcaa gggaaccctc atcaagttga
3901 tttctgccac gttccatacg gtgcgagtca tattgaatgc agtttctccg tctcgttttc
3961 ttctgaacta cgtcaaccat ataagtgtaa ctcaagcaaa gttaaacaaa cgctagtgca
4021 attagtcgag ctctacgaaa cgaaaatcgg ctggactgag ctagcaaccc gatatttgat
4081 gaacatttgc aacggtaaat ggctgtggaa aaatacccgt aaagcctatt gctggaacat
4141 tgtacttaca ccttggcctt ggaacgggga aaaggttgga tttgaagata ttcgtactaa
4201 ctacacctca cggcaagact ttaaaaataa taaaaattgg tctgctatag ttgaaatgat
4261 caaaaccgca ttttctagta ctgatgggct ggcgatattt gaagtcaggg ccaccttgca
4321 cttgccaacg aatgctatgg tgcggccaag ccaagttttc acagaaaaag aaagtggcag
4381 taaaagtaaa tctaaaactc aaaacagtcg agtttttcag agtacaacta ttgatggtga
4441 acgatcgcca atactagggg cctttaaaac gggagcagct attgcaacca ttgacgactg
4501 gtatcctgaa gccactgagc cactaagggt cggacggttt ggggttcatc gcgaagatgt
4561 cacttgctac cgtcatccgt ctaccggaaa agattttttc tcgatattac aacaagcaga
4621 gcactatatt gaagtgttga gcgccaacaa aactcccgct caagaaacta tcaacgacat
4681 gcacttttta atggctaacc tgattaaggg tgggatgttc cagcataaag gagactgact
4741 gtgaaatggt attataagac aatcaccttt ctgccagagt tgtgcaacaa cgagtcactg
4801 gctgcaaagt gtctccgcgt tctgcatgga tttaactatc agtatgagac acgaaatatc
4861 ggcgtttcat ttccgctttg gtgtgatgca acggttggaa aaaagatttc atttgtcagc
4921 aagaacaaga tagaactcga cttactactt aaacaacact atttcgtcca aatggaacaa
4981 cttcaatact ttcatatatc caacactgtt ctcgtcccag aagattgtac atacgtttcc
5041 tttagacgct gtcaatctat agataagctc acagcagcag ggctggcaag gaaaatcaga
5101 cgcctggaga aacgtgctct atctagaggc gagcaatttg acccatcatc ttttgctcaa
5161 aaagagcata ctgcaatagc gcactaccac tcacttgggg agtccagcaa acagacgaac
5221 cgcaactttc gactcaatat caggatgctc tcggagcaac cccgtgaggg gaactcgatt
5281 tttagtagct atggcttatc aaattcagaa aactcgtttc agcctgtacc cttaatctga
5341 cctaggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
5401 tcttgagggg ttttttgctg aaacctcagg catttgagaa gcacacggtc acactgcttc
5461 cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc
5521 ctgaaccgac gaccgggtca tcgtggccgg atcttgcggc ccctcggctt gaacgaattg
5581 ttagacatta tttgccgact accttggtga tctcgccttt cacgtagtgg acaaattctt
5641 ccaactgatc tgcgcgcgag gccaagcgat cttcttcttg tccaagataa gcctgtctag
5701 cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag tcggcagcga
5761 catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac aacgtaagca
5821 ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat
5881 ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc gccgctggac
5941 ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga
6001 tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct ccaaattgca
6061 gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt
6121 ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa aggtcgttga
6181 tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat
6241 cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg gccagcaacg
6301 tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc gatacttcgg
6361 cgatcaccgc ttccctcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
6421 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa atagctagct
6481 cactcggtcg ctacgctccg ggcgtgagac tgcggcgggc gctgcggaca catacaaagt
6541 tacccacaga ttccgtggat aagcagggga ctaacatgtg aggcaaaaca gcagggccgc
6601 gccggtggcg tttttccata ggctccgccc tcctgccaga gttcacataa acagacgctt
6661 ttccggtgca tctgtgggag ccgtgaggct caaccatgaa tctgacagta cgggcgaaac
6721 ccgacaggac ttaaagatcc ccaccgtttc cggcgggtcg ctccctcttg cgctctcctg
6781 ttccgaccct gccgtttacc ggatacctgt tccgcctttc tcccttacgg gaagtgtggc
6841 gctttctcat agctcacaca ctggtatctc ggctcggtgt aggtcgttcg ctccaagctg
6901 ggctgtaagc aagaactccc cgttcagccc gactgctgcg ccttatccgg taactgttca
6961 cttgagtcca acccggaaaa gcacggtaaa acgccactgg cagcagccat tggtaactgg
7021 gagttcgcag aggatttgtt tagctaaaca cgcggttgct cttgaagtgt gcgccaaagt
7081 ccggctacac tggaaggaca gatttggttg ctgtgctctg cgaaagccag ttaccacggt
7141 taagcagttc cccaactgac ttaaccttcg atcaaaccac ctccccaggt ggttttttcg
7201 aaggttaagt cagttgggga actgcttaac cttcgatcaa accacctccc caggtggttt
7261 tttcgtttac agggcaaaag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
7321 tcttttctac tgaaccgctc tagatttcag tgcaatttat ctcttcaaat gtagcacctg
7381 aagtcagccc catacgatat aagttgtaat tctcatgtta gtcatgcccc gcgcccaccg
7441 gaaggagctg actgggttga aggctctcaa gggcatcctg atgaatcccc taatgatttt
7501 tatcaaaatc attaaggtta ccatcacgga aaaaggttat gctgctttta agacccactt
7561 tcacatttaa gttgtttttc taatccgcat atgatcaatt caaggccgaa taagaaggct
7621 ggctctgcac cttggtgatc aaataattcg atagcttgtc gtaataatgg cggcatacta
7681 tcagtagtag gtgtttccct ttcttcttta gcgacttgat gctcttgatc ttccaatacg
7741 caacctaaag taaaatgccc cacagcgctg agtgcatata atgcattctc tagtgaaaaa
7801 ccttgttggc ataaaaaggc taattgattt tcgagagttt catactgttt ttctgtaggc
7861 cgtgtaccta aatgtacttt tgctccatcg cgatgactta gtaaagcaca tctaaaactt
7921 ttagcgttat tacgtaaaaa atcttgccag ctttcccctt ctaaagggca aaagtgagta
7981 tggtgcctat ctaacatctc aatggctaag gcgtcgagca aagcccgctt attttttaca
8041 tgccaataca atgtaggctg ctctacacct agcttctggg cgagtttacg ggttgttaaa
8101 ccttcgattc cgacctcatt aagcagctct aatgcgctgt taatcacttt acttttatct
8161 aatctagaca tcattaattc ctaatttttg ttgacactct atcattgata gagttatttt
8221 accactccct atcagtgata gagaaaagtg aactctagaa ataattttgt ttaactttaa
8281 aaggagatat accatgggtg aactgccgag taggtagctg ataactggat gctttgcgtg
8341 caaaggaacc tgaactcgtg aactgccgag taggtagctg ataacgcctg gcttggcgat
8401 gctcgcctga tcgacaagtg aactgccgag taggtagctg ataacgagaa tattgtggca
8461 aaagcgcata aggtgctg
//
