| 质粒名称: | pQCascade-IS186 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 140627 |
| 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: | 8510 bp |
| 载体类型: | Bacterial Expression, CRISPR |
| 载体抗性: | Spectinomycin |
| 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, crRNA-IS186 |
| 物种来源: | Vibrio cholera |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | Unknown (unknown if destroyed) |
| 3’克隆位点: | Unknown (unknown if destroyed) |
|
文献引用方法 |
|
| 材料和方法部分: | pQCascade-IS186 was a gift from Sheng Yang (Addgene plasmid # 140627 ; http://n2t.net/addgene:140627 ; RRID:Addgene_140627) |
| 参考文献部分: | Multicopy chromosomal integration using CRISPR-associated transposases. Zhang Y, Sun X, Wang Q, Xu J, Dong F, Yang S, Yang J, Zhang Z, Qian Y, Chen J, Zhang J, Liu Y, Tao R, Jiang Y, Yang J, Yang S. ACS Synth Biol. 2020 Jun 17. doi: 10.1021/acssynbio.0c00073. 10.1021/acssynbio.0c00073 PubMed 32551502 |
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 pQCascade-IS186 8510 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses V. cholerae TniQ, Cas8, Cas7, and Cas6 and crRNA-IS186.
The crRNA-IS186 targets IS186 loci in Escherchia coli..
ACCESSION .
VERSION .
KEYWORDS pQCascade-IS186
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8510)
AUTHORS Zhang Y, Sun X, Wang Q, Xu J, Dong F, Yang S, Yang J, Zhang Z, Qian
Y, Chen J, Zhang J, Liu Y, Tao R, Jiang Y, Yang J, Yang S
TITLE Multicopy chromosomal integration using CRISPR-associated
transposases.
JOURNAL ACS Synth Biol. 2020 Jun 17. doi: 10.1021/acssynbio.0c00073.
PUBMED 32551502
REFERENCE 2 (bases 1 to 8510)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8510
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 970..989
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(1009..1086)
/gene="lacI"
/label=lacI promoter
promoter 1132..1150
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1151..1175
/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)."
promoter 1419..1437
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1438..1462
/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 complement(6260..6278)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 6274..6321
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(6492..7283)
/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(7284..7375)
/gene="bla"
/label=AmpR promoter
CDS complement(8436..1008)
/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"
ORIGIN
1 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
61 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
121 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
181 ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat
241 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
301 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
361 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
421 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
481 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
541 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
601 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
661 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
721 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
781 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
841 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
901 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
961 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
1021 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
1081 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaaa ttaatacgac
1141 tcactatagg ggaattgtga gcggataaca attcccctgt agaaataatt ttgtttaact
1201 ttaataagga gatataccat gggtgaactg ccgagtaggt agctgataac tggatgcttt
1261 gcgtgcaaag gaacctgaac tcgtgaactg ccgagtaggt agctgataac ggatccgaat
1321 tcgagctcgg cgcgcctgca ggtcgacaag cttgcggccg cataatgctt aagtcgaaca
1381 gaaagtaatc gtattgtaca cggccgcata atcgaaatta atacgactca ctatagggga
1441 attgtgagcg gataacaatt ccccatctta gtatattagt taagtataag aaggagatat
1501 acatatgttt ttgcaaagac ctaaacctta cagcgatgaa agtttagaaa gtttctttat
1561 ccgagtggct aacaaaaatg gctacggtga tgtccatcgc ttcctagaag ccactaaacg
1621 attccttcaa gacattgacc ataatggcta tcaaaccttt ccgactgata taactcggat
1681 aaacccatac tcagctaaaa acagttccag cgcacgaact gcgtcattcc tgaaacttgc
1741 acaattgaca tttaatgaac cgccagagct acttgggttg gcaattaaca gaacaaacat
1801 gaaatactcg ccgtcaacta gcgcggttgt tagaggtgca gaagtctttc ctcgcagttt
1861 actacggacg cactccatcc cctgctgtcc tttgtgtctg cgagaaaatg gctacgcctc
1921 ctacctttgg cactttcagg ggtacgaata ctgccacagc cataacgtac ctttaattac
1981 cacttgtagc tgtggtaagg agtttgacta ccgagtatct gggttaaagg gcatttgctg
2041 caaatgcaag gagcctatca ccttaaccag cagggagaac ggtcatgagg cagcgtgtac
2101 tgtttcaaac tggcttgctg gccatgaatc taaacctctg ccaaatcttc ctaaaagcta
2161 ccgatggggt ttagttcatt ggtggatggg tattaaagat agcgagttcg atcacttttc
2221 gttcgttcaa tttttctcaa actggccaag gtcattccac tcgataatcg aagatgaagt
2281 agagttcaac cttgagcatg ctgttgtcag cacgtctgaa ttacgactaa aagatcttct
2341 tggtcgattg tttttcggtt caattcggtt acctgagcgg aatcttcaac acaatatcat
2401 ccttggtgag cttctctgct atttagaaaa tcgtttatgg caagacaagg gattaatcgc
2461 caacctcaaa atgaacgcgt tagaggcgac tgtaatgtta aattgtagcc tcgatcagat
2521 tgcatcaatg gttgaacaac gcatcttgaa gccaaatcga aaaagcaagc ccaacagccc
2581 tcttgatgtt accgattatc tatttcattt cggcgatatt ttctgtcttt ggttagctga
2641 gttccaaagc gatgagttta accgttcgtt ttatgtgtcg aggtggtaaa tgcaaactct
2701 gaaagaacta atcgcatcca atcctgacga cttaacaact gaacttaaga gagcatttcg
2761 tccactcaca ccgcatattg caattgatgg taatgaactt gacgcactga cgatattagt
2821 caatttaacc gataagactg atgatcagaa agacctgctc gatcgagcca aatgcaagca
2881 aaaacttcga gatgaaaaat ggtgggctag ctgcataaat tgcgttaatt acagacaaag
2941 ccataaccca aaattcccgg atatacgttc tgaaggcgtg attcgaaccc aagccctggg
3001 tgaattaccg agtttcctac tctcgtcctc aaaaatccca ccataccatt ggtcatatag
3061 ccatgattca aaatacgtca acaaaagcgc attcctcacc aatgagtttt gttgggatgg
3121 tgagatctca tgtttgggtg agcttcttaa agatgcagat cacccacttt ggaatacttt
3181 gaaaaagtta ggttgttctc aaaaaacttg caaagcaatg gcaaaacaac tagctgatat
3241 tactctcacg actatcaatg tcacactcgc accaaattac ctgactcaaa tatctctccc
3301 cgatagtgat acatcctaca tttcactctc tcctgttgca tcgctatcga tgcaaagcca
3361 ctttcatcag aggctccaag atgaaaatcg gcatagcgcg ataacgcggt ttagccgaac
3421 taccaacatg ggggtcactg cgatgacatg tggcggcgca tttagaatgt taaagtctgg
3481 cgctaagttt tccagccccc ctcatcaccg attaaacagt aaacgaagtt ggttgacgtc
3541 agagcatgtt cagtcattaa aacagtacca gcgcctcaac aaaagcctca tacctgaaaa
3601 ctctcggatt gcactccgta gaaaatacaa aatcgagctt caaaatatgg tcagatcttg
3661 gtttgcaatg caagaccata cccttgattc gaacatactt atccaacatt tgaatcatga
3721 cctatcttac ttaggagcca caaaacgttt tgcatacgat ccagcgatga ccaagctctt
3781 tactgagctt ttgaaacgag agttatcaaa ttcaatcaat aatggtgagc aacacactaa
3841 tggatcgttt ttagtcctac cgaatatcag ggtttgtggc gcaacagctt taagctcccc
3901 ggtaacggtg gggattccat cacttacagc tttctttggc ttcgttcacg catttgaacg
3961 gaatataaat cgcaccacct catcgtttcg tgttgaatcc tttgcgatat gcgtccatca
4021 actacatgtc gagaagcgag gtttgacagc agagtttgtg gaaaaaggcg acgggactat
4081 atccgctccc gcgacccggg atgactggca gtgtgatgtc gtatttagcc ttattttgaa
4141 caccaacttt gctcaacata ttgaccaaga tacgttagtt acatcactac caaagcgatt
4201 ggctcggggt tcagcaaaaa ttgcgattga tgactttaaa catatcaact cattctcgac
4261 attagaaaca gcgatcgaat ctctgccaat agaagctggt aggtggttat cactttacgc
4321 acagtcaaac aataatctaa gtgatctatt agcagccatg acagaggacc atcagctcat
4381 ggcaagctgc gtcggttacc acttgttaga agagcccaaa gataaaccaa actccctcag
4441 aggttacaaa cacgctatcg ccgagtgcat cattggactc attaactcaa tcacctttag
4501 ctcagagact gatcccaaca caatcttttg gtcgctaaag aactatcaaa actacctagt
4561 ggtacagcca aggagtatca acgatgaaac taccgacaaa tctagcctat gagcgctcta
4621 tcgacccatc agatgtctgt ttttttgtcg tctggcccga tgatagaaaa acacctttaa
4681 cctacaattc tcgtactctg ctcgggcaaa tggaagcggc atcattagcc tatgatgtct
4741 caggtcaacc aataaaaagt gccaccgctg aggcgttagc tcaagggaac cctcatcaag
4801 ttgatttctg ccacgttcca tacggtgcga gtcatattga atgcagtttc tccgtctcgt
4861 tttcttctga actacgtcaa ccatataagt gtaactcaag caaagttaaa caaacgctag
4921 tgcaattagt cgagctctac gaaacgaaaa tcggctggac tgagctagca acccgatatt
4981 tgatgaacat ttgcaacggt aaatggctgt ggaaaaatac ccgtaaagcc tattgctgga
5041 acattgtact tacaccttgg ccttggaacg gggaaaaggt tggatttgaa gatattcgta
5101 ctaactacac ctcacggcaa gactttaaaa ataataaaaa ttggtctgct atagttgaaa
5161 tgatcaaaac cgcattttct agtactgatg ggctggcgat atttgaagtc agggccacct
5221 tgcacttgcc aacgaatgct atggtgcggc caagccaagt tttcacagaa aaagaaagtg
5281 gcagtaaaag taaatctaaa actcaaaaca gtcgagtttt tcagagtaca actattgatg
5341 gtgaacgatc gccaatacta ggggccttta aaacgggagc agctattgca accattgacg
5401 actggtatcc tgaagccact gagccactaa gggtcggacg gtttggggtt catcgcgaag
5461 atgtcacttg ctaccgtcat ccgtctaccg gaaaagattt tttctcgata ttacaacaag
5521 cagagcacta tattgaagtg ttgagcgcca acaaaactcc cgctcaagaa actatcaacg
5581 acatgcactt tttaatggct aacctgatta agggtgggat gttccagcat aaaggagact
5641 gactgtgaaa tggtattata agacaatcac ctttctgcca gagttgtgca acaacgagtc
5701 actggctgca aagtgtctcc gcgttctgca tggatttaac tatcagtatg agacacgaaa
5761 tatcggcgtt tcatttccgc tttggtgtga tgcaacggtt ggaaaaaaga tttcatttgt
5821 cagcaagaac aagatagaac tcgacttact acttaaacaa cactatttcg tccaaatgga
5881 acaacttcaa tactttcata tatccaacac tgttctcgtc ccagaagatt gtacatacgt
5941 ttcctttaga cgctgtcaat ctatagataa gctcacagca gcagggctgg caaggaaaat
6001 cagacgcctg gagaaacgtg ctctatctag aggcgagcaa tttgacccat catcttttgc
6061 tcaaaaagag catactgcaa tagcgcacta ccactcactt ggggagtcca gcaaacagac
6121 gaaccgcaac tttcgactca atatcaggat gctctcggag caaccccgtg aggggaactc
6181 gatttttagt agctatggct tatcaaattc agaaaactcg tttcagcctg tacccttaat
6241 ctgacctagg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
6301 cgggtcttga ggggtttttt gctgaaacct caggcatttg agaagcacac ggtcacactg
6361 cttccggtag tcaataaacc ggtaaaccag caatagacat aagcggctat ttaacgaccc
6421 tgccctgaac cgacgaccgg gtcatcgtgg ccggatcttg cggcccctcg gcttgaacga
6481 attgttagac attatttgcc gactaccttg gtgatctcgc ctttcacgta gtggacaaat
6541 tcttccaact gatctgcgcg cgaggccaag cgatcttctt cttgtccaag ataagcctgt
6601 ctagcttcaa gtatgacggg ctgatactgg gccggcaggc gctccattgc ccagtcggca
6661 gcgacatcct tcggcgcgat tttgccggtt actgcgctgt accaaatgcg ggacaacgta
6721 agcactacat ttcgctcatc gccagcccag tcgggcggcg agttccatag cgttaaggtt
6781 tcatttagcg cctcaaatag atcctgttca ggaaccggat caaagagttc ctccgccgct
6841 ggacctacca aggcaacgct atgttctctt gcttttgtca gcaagatagc cagatcaatg
6901 tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat tgcgctgcca ttctccaaat
6961 tgcagttcgc gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg
7021 acttctacag cgcggagaat ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg
7081 ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac cagcaaatca
7141 atatcactgt gtggcttcag gccgccatcc actgcggagc cgtacaaatg tacggccagc
7201 aacgtcggtt cgagatggcg ctcgatgacg ccaactacct ctgatagttg agtcgatact
7261 tcggcgatca ccgcttccct catactcttc ctttttcaat attattgaag catttatcag
7321 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaatagct
7381 agctcactcg gtcgctacgc tccgggcgtg agactgcggc gggcgctgcg gacacataca
7441 aagttaccca cagattccgt ggataagcag gggactaaca tgtgaggcaa aacagcaggg
7501 ccgcgccggt ggcgtttttc cataggctcc gccctcctgc cagagttcac ataaacagac
7561 gcttttccgg tgcatctgtg ggagccgtga ggctcaacca tgaatctgac agtacgggcg
7621 aaacccgaca ggacttaaag atccccaccg tttccggcgg gtcgctccct cttgcgctct
7681 cctgttccga ccctgccgtt taccggatac ctgttccgcc tttctccctt acgggaagtg
7741 tggcgctttc tcatagctca cacactggta tctcggctcg gtgtaggtcg ttcgctccaa
7801 gctgggctgt aagcaagaac tccccgttca gcccgactgc tgcgccttat ccggtaactg
7861 ttcacttgag tccaacccgg aaaagcacgg taaaacgcca ctggcagcag ccattggtaa
7921 ctgggagttc gcagaggatt tgtttagcta aacacgcggt tgctcttgaa gtgtgcgcca
7981 aagtccggct acactggaag gacagatttg gttgctgtgc tctgcgaaag ccagttacca
8041 cggttaagca gttccccaac tgacttaacc ttcgatcaaa ccacctcccc aggtggtttt
8101 ttcgaaggtt aagtcagttg gggaactgct taaccttcga tcaaaccacc tccccaggtg
8161 gttttttcgt ttacagggca aaagattacg cgcagaaaaa aaggatctca agaagatcct
8221 ttgatctttt ctactgaacc gctctagatt tcagtgcaat ttatctcttc aaatgtagca
8281 cctgaagtca gccccatacg atataagttg taattctcat gttagtcatg ccccgcgccc
8341 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct
8401 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
8461 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
//
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