| 质粒名称: | pQCascade-IS3 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 140626 |
| 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 ; Transposon |
| 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-IS3 |
| 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-IS3 was a gift from Sheng Yang (Addgene plasmid # 140626 ; http://n2t.net/addgene:140626 ; RRID:Addgene_140626) |
| 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
pQCascade-IS3载体质粒基本信息
pQCascade-IS3质粒图谱载体图谱和pQCascade-IS3载体序列质粒序列多克隆位点信息
pQCascade-IS3质粒载体简介
pQCascade-IS3质粒序列载体序列
LOCUS pQCascade-IS3 8510 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Expresses V. cholerae TniQ, Cas8, Cas7, and Cas6 and crRNA-IS3. The
crRNA-IS3 targets IS3 loci in Escherchia coli..
ACCESSION .
VERSION .
KEYWORDS pQCascade-IS3
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 13, 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"
promoter 119..137
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 138..162
/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(4960..4978)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 4974..5021
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(5192..5983)
/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(5984..6075)
/gene="bla"
/label=AmpR promoter
CDS complement(7136..8218)
/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"
primer_bind 8180..8199
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(8219..8296)
/gene="lacI"
/label=lacI promoter
promoter 8342..8360
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 8361..8385
/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)."
ORIGIN
1 agctgataac ggatccgaat tcgagctcgg cgcgcctgca ggtcgacaag cttgcggccg
61 cataatgctt aagtcgaaca gaaagtaatc gtattgtaca cggccgcata atcgaaatta
121 atacgactca ctatagggga attgtgagcg gataacaatt ccccatctta gtatattagt
181 taagtataag aaggagatat acatatgttt ttgcaaagac ctaaacctta cagcgatgaa
241 agtttagaaa gtttctttat ccgagtggct aacaaaaatg gctacggtga tgtccatcgc
301 ttcctagaag ccactaaacg attccttcaa gacattgacc ataatggcta tcaaaccttt
361 ccgactgata taactcggat aaacccatac tcagctaaaa acagttccag cgcacgaact
421 gcgtcattcc tgaaacttgc acaattgaca tttaatgaac cgccagagct acttgggttg
481 gcaattaaca gaacaaacat gaaatactcg ccgtcaacta gcgcggttgt tagaggtgca
541 gaagtctttc ctcgcagttt actacggacg cactccatcc cctgctgtcc tttgtgtctg
601 cgagaaaatg gctacgcctc ctacctttgg cactttcagg ggtacgaata ctgccacagc
661 cataacgtac ctttaattac cacttgtagc tgtggtaagg agtttgacta ccgagtatct
721 gggttaaagg gcatttgctg caaatgcaag gagcctatca ccttaaccag cagggagaac
781 ggtcatgagg cagcgtgtac tgtttcaaac tggcttgctg gccatgaatc taaacctctg
841 ccaaatcttc ctaaaagcta ccgatggggt ttagttcatt ggtggatggg tattaaagat
901 agcgagttcg atcacttttc gttcgttcaa tttttctcaa actggccaag gtcattccac
961 tcgataatcg aagatgaagt agagttcaac cttgagcatg ctgttgtcag cacgtctgaa
1021 ttacgactaa aagatcttct tggtcgattg tttttcggtt caattcggtt acctgagcgg
1081 aatcttcaac acaatatcat ccttggtgag cttctctgct atttagaaaa tcgtttatgg
1141 caagacaagg gattaatcgc caacctcaaa atgaacgcgt tagaggcgac tgtaatgtta
1201 aattgtagcc tcgatcagat tgcatcaatg gttgaacaac gcatcttgaa gccaaatcga
1261 aaaagcaagc ccaacagccc tcttgatgtt accgattatc tatttcattt cggcgatatt
1321 ttctgtcttt ggttagctga gttccaaagc gatgagttta accgttcgtt ttatgtgtcg
1381 aggtggtaaa tgcaaactct gaaagaacta atcgcatcca atcctgacga cttaacaact
1441 gaacttaaga gagcatttcg tccactcaca ccgcatattg caattgatgg taatgaactt
1501 gacgcactga cgatattagt caatttaacc gataagactg atgatcagaa agacctgctc
1561 gatcgagcca aatgcaagca aaaacttcga gatgaaaaat ggtgggctag ctgcataaat
1621 tgcgttaatt acagacaaag ccataaccca aaattcccgg atatacgttc tgaaggcgtg
1681 attcgaaccc aagccctggg tgaattaccg agtttcctac tctcgtcctc aaaaatccca
1741 ccataccatt ggtcatatag ccatgattca aaatacgtca acaaaagcgc attcctcacc
1801 aatgagtttt gttgggatgg tgagatctca tgtttgggtg agcttcttaa agatgcagat
1861 cacccacttt ggaatacttt gaaaaagtta ggttgttctc aaaaaacttg caaagcaatg
1921 gcaaaacaac tagctgatat tactctcacg actatcaatg tcacactcgc accaaattac
1981 ctgactcaaa tatctctccc cgatagtgat acatcctaca tttcactctc tcctgttgca
2041 tcgctatcga tgcaaagcca ctttcatcag aggctccaag atgaaaatcg gcatagcgcg
2101 ataacgcggt ttagccgaac taccaacatg ggggtcactg cgatgacatg tggcggcgca
2161 tttagaatgt taaagtctgg cgctaagttt tccagccccc ctcatcaccg attaaacagt
2221 aaacgaagtt ggttgacgtc agagcatgtt cagtcattaa aacagtacca gcgcctcaac
2281 aaaagcctca tacctgaaaa ctctcggatt gcactccgta gaaaatacaa aatcgagctt
2341 caaaatatgg tcagatcttg gtttgcaatg caagaccata cccttgattc gaacatactt
2401 atccaacatt tgaatcatga cctatcttac ttaggagcca caaaacgttt tgcatacgat
2461 ccagcgatga ccaagctctt tactgagctt ttgaaacgag agttatcaaa ttcaatcaat
2521 aatggtgagc aacacactaa tggatcgttt ttagtcctac cgaatatcag ggtttgtggc
2581 gcaacagctt taagctcccc ggtaacggtg gggattccat cacttacagc tttctttggc
2641 ttcgttcacg catttgaacg gaatataaat cgcaccacct catcgtttcg tgttgaatcc
2701 tttgcgatat gcgtccatca actacatgtc gagaagcgag gtttgacagc agagtttgtg
2761 gaaaaaggcg acgggactat atccgctccc gcgacccggg atgactggca gtgtgatgtc
2821 gtatttagcc ttattttgaa caccaacttt gctcaacata ttgaccaaga tacgttagtt
2881 acatcactac caaagcgatt ggctcggggt tcagcaaaaa ttgcgattga tgactttaaa
2941 catatcaact cattctcgac attagaaaca gcgatcgaat ctctgccaat agaagctggt
3001 aggtggttat cactttacgc acagtcaaac aataatctaa gtgatctatt agcagccatg
3061 acagaggacc atcagctcat ggcaagctgc gtcggttacc acttgttaga agagcccaaa
3121 gataaaccaa actccctcag aggttacaaa cacgctatcg ccgagtgcat cattggactc
3181 attaactcaa tcacctttag ctcagagact gatcccaaca caatcttttg gtcgctaaag
3241 aactatcaaa actacctagt ggtacagcca aggagtatca acgatgaaac taccgacaaa
3301 tctagcctat gagcgctcta tcgacccatc agatgtctgt ttttttgtcg tctggcccga
3361 tgatagaaaa acacctttaa cctacaattc tcgtactctg ctcgggcaaa tggaagcggc
3421 atcattagcc tatgatgtct caggtcaacc aataaaaagt gccaccgctg aggcgttagc
3481 tcaagggaac cctcatcaag ttgatttctg ccacgttcca tacggtgcga gtcatattga
3541 atgcagtttc tccgtctcgt tttcttctga actacgtcaa ccatataagt gtaactcaag
3601 caaagttaaa caaacgctag tgcaattagt cgagctctac gaaacgaaaa tcggctggac
3661 tgagctagca acccgatatt tgatgaacat ttgcaacggt aaatggctgt ggaaaaatac
3721 ccgtaaagcc tattgctgga acattgtact tacaccttgg ccttggaacg gggaaaaggt
3781 tggatttgaa gatattcgta ctaactacac ctcacggcaa gactttaaaa ataataaaaa
3841 ttggtctgct atagttgaaa tgatcaaaac cgcattttct agtactgatg ggctggcgat
3901 atttgaagtc agggccacct tgcacttgcc aacgaatgct atggtgcggc caagccaagt
3961 tttcacagaa aaagaaagtg gcagtaaaag taaatctaaa actcaaaaca gtcgagtttt
4021 tcagagtaca actattgatg gtgaacgatc gccaatacta ggggccttta aaacgggagc
4081 agctattgca accattgacg actggtatcc tgaagccact gagccactaa gggtcggacg
4141 gtttggggtt catcgcgaag atgtcacttg ctaccgtcat ccgtctaccg gaaaagattt
4201 tttctcgata ttacaacaag cagagcacta tattgaagtg ttgagcgcca acaaaactcc
4261 cgctcaagaa actatcaacg acatgcactt tttaatggct aacctgatta agggtgggat
4321 gttccagcat aaaggagact gactgtgaaa tggtattata agacaatcac ctttctgcca
4381 gagttgtgca acaacgagtc actggctgca aagtgtctcc gcgttctgca tggatttaac
4441 tatcagtatg agacacgaaa tatcggcgtt tcatttccgc tttggtgtga tgcaacggtt
4501 ggaaaaaaga tttcatttgt cagcaagaac aagatagaac tcgacttact acttaaacaa
4561 cactatttcg tccaaatgga acaacttcaa tactttcata tatccaacac tgttctcgtc
4621 ccagaagatt gtacatacgt ttcctttaga cgctgtcaat ctatagataa gctcacagca
4681 gcagggctgg caaggaaaat cagacgcctg gagaaacgtg ctctatctag aggcgagcaa
4741 tttgacccat catcttttgc tcaaaaagag catactgcaa tagcgcacta ccactcactt
4801 ggggagtcca gcaaacagac gaaccgcaac tttcgactca atatcaggat gctctcggag
4861 caaccccgtg aggggaactc gatttttagt agctatggct tatcaaattc agaaaactcg
4921 tttcagcctg tacccttaat ctgacctagg ctgctgccac cgctgagcaa taactagcat
4981 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaacct caggcatttg
5041 agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag caatagacat
5101 aagcggctat ttaacgaccc tgccctgaac cgacgaccgg gtcatcgtgg ccggatcttg
5161 cggcccctcg gcttgaacga attgttagac attatttgcc gactaccttg gtgatctcgc
5221 ctttcacgta gtggacaaat tcttccaact gatctgcgcg cgaggccaag cgatcttctt
5281 cttgtccaag ataagcctgt ctagcttcaa gtatgacggg ctgatactgg gccggcaggc
5341 gctccattgc ccagtcggca gcgacatcct tcggcgcgat tttgccggtt actgcgctgt
5401 accaaatgcg ggacaacgta agcactacat ttcgctcatc gccagcccag tcgggcggcg
5461 agttccatag cgttaaggtt tcatttagcg cctcaaatag atcctgttca ggaaccggat
5521 caaagagttc ctccgccgct ggacctacca aggcaacgct atgttctctt gcttttgtca
5581 gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat
5641 tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg ataacgccac ggaatgatgt
5701 cgtcgtgcac aacaatggtg acttctacag cgcggagaat ctcgctctct ccaggggaag
5761 ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg
5821 tcaccgtaac cagcaaatca atatcactgt gtggcttcag gccgccatcc actgcggagc
5881 cgtacaaatg tacggccagc aacgtcggtt cgagatggcg ctcgatgacg ccaactacct
5941 ctgatagttg agtcgatact tcggcgatca ccgcttccct catactcttc ctttttcaat
6001 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
6061 agaaaaataa acaaatagct agctcactcg gtcgctacgc tccgggcgtg agactgcggc
6121 gggcgctgcg gacacataca aagttaccca cagattccgt ggataagcag gggactaaca
6181 tgtgaggcaa aacagcaggg ccgcgccggt ggcgtttttc cataggctcc gccctcctgc
6241 cagagttcac ataaacagac gcttttccgg tgcatctgtg ggagccgtga ggctcaacca
6301 tgaatctgac agtacgggcg aaacccgaca ggacttaaag atccccaccg tttccggcgg
6361 gtcgctccct cttgcgctct cctgttccga ccctgccgtt taccggatac ctgttccgcc
6421 tttctccctt acgggaagtg tggcgctttc tcatagctca cacactggta tctcggctcg
6481 gtgtaggtcg ttcgctccaa gctgggctgt aagcaagaac tccccgttca gcccgactgc
6541 tgcgccttat ccggtaactg ttcacttgag tccaacccgg aaaagcacgg taaaacgcca
6601 ctggcagcag ccattggtaa ctgggagttc gcagaggatt tgtttagcta aacacgcggt
6661 tgctcttgaa gtgtgcgcca aagtccggct acactggaag gacagatttg gttgctgtgc
6721 tctgcgaaag ccagttacca cggttaagca gttccccaac tgacttaacc ttcgatcaaa
6781 ccacctcccc aggtggtttt ttcgaaggtt aagtcagttg gggaactgct taaccttcga
6841 tcaaaccacc tccccaggtg gttttttcgt ttacagggca aaagattacg cgcagaaaaa
6901 aaggatctca agaagatcct ttgatctttt ctactgaacc gctctagatt tcagtgcaat
6961 ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat
7021 gttagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
7081 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
7141 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
7201 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
7261 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
7321 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
7381 agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg
7441 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
7501 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
7561 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
7621 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
7681 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
7741 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
7801 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
7861 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
7921 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
7981 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
8041 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
8101 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
8161 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
8221 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
8281 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaaa
8341 ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctgt agaaataatt
8401 ttgtttaact ttaataagga gatataccat gggtgaactg ccgagtaggt agctgataac
8461 ggaacgttat cgttcacacg gaccgtggag gcgtgaactg ccgagtaggt
//
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