| 质粒名称: | pQCascade-IS2 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 140625 |
| 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: | 8510bp |
| 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-IS2 |
| 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-IS2 was a gift from Sheng Yang (Addgene plasmid # 140625 ; http://n2t.net/addgene:140625 ; RRID:Addgene_140625) |
| 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-IS2 8510 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Expresses V. cholerae TniQ, Cas8, Cas7, and Cas6 and crRNA-IS2. The
crRNA-IS2 targets IS2 loci in Escherchia coli..
ACCESSION .
VERSION .
KEYWORDS pQCascade-IS2
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 166..184
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 185..209
/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(5007..5025)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 5021..5068
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(5239..6030)
/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(6031..6122)
/gene="bla"
/label=AmpR promoter
CDS complement(7183..8265)
/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 8227..8246
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(8266..8343)
/gene="lacI"
/label=lacI promoter
promoter 8389..8407
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 8408..8432
/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 cagaggcgtt cgagcattat aacgaatggg tgaactgccg agtaggtagc tgataacgga
61 tccgaattcg agctcggcgc gcctgcaggt cgacaagctt gcggccgcat aatgcttaag
121 tcgaacagaa agtaatcgta ttgtacacgg ccgcataatc gaaattaata cgactcacta
181 taggggaatt gtgagcggat aacaattccc catcttagta tattagttaa gtataagaag
241 gagatataca tatgtttttg caaagaccta aaccttacag cgatgaaagt ttagaaagtt
301 tctttatccg agtggctaac aaaaatggct acggtgatgt ccatcgcttc ctagaagcca
361 ctaaacgatt ccttcaagac attgaccata atggctatca aacctttccg actgatataa
421 ctcggataaa cccatactca gctaaaaaca gttccagcgc acgaactgcg tcattcctga
481 aacttgcaca attgacattt aatgaaccgc cagagctact tgggttggca attaacagaa
541 caaacatgaa atactcgccg tcaactagcg cggttgttag aggtgcagaa gtctttcctc
601 gcagtttact acggacgcac tccatcccct gctgtccttt gtgtctgcga gaaaatggct
661 acgcctccta cctttggcac tttcaggggt acgaatactg ccacagccat aacgtacctt
721 taattaccac ttgtagctgt ggtaaggagt ttgactaccg agtatctggg ttaaagggca
781 tttgctgcaa atgcaaggag cctatcacct taaccagcag ggagaacggt catgaggcag
841 cgtgtactgt ttcaaactgg cttgctggcc atgaatctaa acctctgcca aatcttccta
901 aaagctaccg atggggttta gttcattggt ggatgggtat taaagatagc gagttcgatc
961 acttttcgtt cgttcaattt ttctcaaact ggccaaggtc attccactcg ataatcgaag
1021 atgaagtaga gttcaacctt gagcatgctg ttgtcagcac gtctgaatta cgactaaaag
1081 atcttcttgg tcgattgttt ttcggttcaa ttcggttacc tgagcggaat cttcaacaca
1141 atatcatcct tggtgagctt ctctgctatt tagaaaatcg tttatggcaa gacaagggat
1201 taatcgccaa cctcaaaatg aacgcgttag aggcgactgt aatgttaaat tgtagcctcg
1261 atcagattgc atcaatggtt gaacaacgca tcttgaagcc aaatcgaaaa agcaagccca
1321 acagccctct tgatgttacc gattatctat ttcatttcgg cgatattttc tgtctttggt
1381 tagctgagtt ccaaagcgat gagtttaacc gttcgtttta tgtgtcgagg tggtaaatgc
1441 aaactctgaa agaactaatc gcatccaatc ctgacgactt aacaactgaa cttaagagag
1501 catttcgtcc actcacaccg catattgcaa ttgatggtaa tgaacttgac gcactgacga
1561 tattagtcaa tttaaccgat aagactgatg atcagaaaga cctgctcgat cgagccaaat
1621 gcaagcaaaa acttcgagat gaaaaatggt gggctagctg cataaattgc gttaattaca
1681 gacaaagcca taacccaaaa ttcccggata tacgttctga aggcgtgatt cgaacccaag
1741 ccctgggtga attaccgagt ttcctactct cgtcctcaaa aatcccacca taccattggt
1801 catatagcca tgattcaaaa tacgtcaaca aaagcgcatt cctcaccaat gagttttgtt
1861 gggatggtga gatctcatgt ttgggtgagc ttcttaaaga tgcagatcac ccactttgga
1921 atactttgaa aaagttaggt tgttctcaaa aaacttgcaa agcaatggca aaacaactag
1981 ctgatattac tctcacgact atcaatgtca cactcgcacc aaattacctg actcaaatat
2041 ctctccccga tagtgataca tcctacattt cactctctcc tgttgcatcg ctatcgatgc
2101 aaagccactt tcatcagagg ctccaagatg aaaatcggca tagcgcgata acgcggttta
2161 gccgaactac caacatgggg gtcactgcga tgacatgtgg cggcgcattt agaatgttaa
2221 agtctggcgc taagttttcc agcccccctc atcaccgatt aaacagtaaa cgaagttggt
2281 tgacgtcaga gcatgttcag tcattaaaac agtaccagcg cctcaacaaa agcctcatac
2341 ctgaaaactc tcggattgca ctccgtagaa aatacaaaat cgagcttcaa aatatggtca
2401 gatcttggtt tgcaatgcaa gaccataccc ttgattcgaa catacttatc caacatttga
2461 atcatgacct atcttactta ggagccacaa aacgttttgc atacgatcca gcgatgacca
2521 agctctttac tgagcttttg aaacgagagt tatcaaattc aatcaataat ggtgagcaac
2581 acactaatgg atcgttttta gtcctaccga atatcagggt ttgtggcgca acagctttaa
2641 gctccccggt aacggtgggg attccatcac ttacagcttt ctttggcttc gttcacgcat
2701 ttgaacggaa tataaatcgc accacctcat cgtttcgtgt tgaatccttt gcgatatgcg
2761 tccatcaact acatgtcgag aagcgaggtt tgacagcaga gtttgtggaa aaaggcgacg
2821 ggactatatc cgctcccgcg acccgggatg actggcagtg tgatgtcgta tttagcctta
2881 ttttgaacac caactttgct caacatattg accaagatac gttagttaca tcactaccaa
2941 agcgattggc tcggggttca gcaaaaattg cgattgatga ctttaaacat atcaactcat
3001 tctcgacatt agaaacagcg atcgaatctc tgccaataga agctggtagg tggttatcac
3061 tttacgcaca gtcaaacaat aatctaagtg atctattagc agccatgaca gaggaccatc
3121 agctcatggc aagctgcgtc ggttaccact tgttagaaga gcccaaagat aaaccaaact
3181 ccctcagagg ttacaaacac gctatcgccg agtgcatcat tggactcatt aactcaatca
3241 cctttagctc agagactgat cccaacacaa tcttttggtc gctaaagaac tatcaaaact
3301 acctagtggt acagccaagg agtatcaacg atgaaactac cgacaaatct agcctatgag
3361 cgctctatcg acccatcaga tgtctgtttt tttgtcgtct ggcccgatga tagaaaaaca
3421 cctttaacct acaattctcg tactctgctc gggcaaatgg aagcggcatc attagcctat
3481 gatgtctcag gtcaaccaat aaaaagtgcc accgctgagg cgttagctca agggaaccct
3541 catcaagttg atttctgcca cgttccatac ggtgcgagtc atattgaatg cagtttctcc
3601 gtctcgtttt cttctgaact acgtcaacca tataagtgta actcaagcaa agttaaacaa
3661 acgctagtgc aattagtcga gctctacgaa acgaaaatcg gctggactga gctagcaacc
3721 cgatatttga tgaacatttg caacggtaaa tggctgtgga aaaatacccg taaagcctat
3781 tgctggaaca ttgtacttac accttggcct tggaacgggg aaaaggttgg atttgaagat
3841 attcgtacta actacacctc acggcaagac tttaaaaata ataaaaattg gtctgctata
3901 gttgaaatga tcaaaaccgc attttctagt actgatgggc tggcgatatt tgaagtcagg
3961 gccaccttgc acttgccaac gaatgctatg gtgcggccaa gccaagtttt cacagaaaaa
4021 gaaagtggca gtaaaagtaa atctaaaact caaaacagtc gagtttttca gagtacaact
4081 attgatggtg aacgatcgcc aatactaggg gcctttaaaa cgggagcagc tattgcaacc
4141 attgacgact ggtatcctga agccactgag ccactaaggg tcggacggtt tggggttcat
4201 cgcgaagatg tcacttgcta ccgtcatccg tctaccggaa aagatttttt ctcgatatta
4261 caacaagcag agcactatat tgaagtgttg agcgccaaca aaactcccgc tcaagaaact
4321 atcaacgaca tgcacttttt aatggctaac ctgattaagg gtgggatgtt ccagcataaa
4381 ggagactgac tgtgaaatgg tattataaga caatcacctt tctgccagag ttgtgcaaca
4441 acgagtcact ggctgcaaag tgtctccgcg ttctgcatgg atttaactat cagtatgaga
4501 cacgaaatat cggcgtttca tttccgcttt ggtgtgatgc aacggttgga aaaaagattt
4561 catttgtcag caagaacaag atagaactcg acttactact taaacaacac tatttcgtcc
4621 aaatggaaca acttcaatac tttcatatat ccaacactgt tctcgtccca gaagattgta
4681 catacgtttc ctttagacgc tgtcaatcta tagataagct cacagcagca gggctggcaa
4741 ggaaaatcag acgcctggag aaacgtgctc tatctagagg cgagcaattt gacccatcat
4801 cttttgctca aaaagagcat actgcaatag cgcactacca ctcacttggg gagtccagca
4861 aacagacgaa ccgcaacttt cgactcaata tcaggatgct ctcggagcaa ccccgtgagg
4921 ggaactcgat ttttagtagc tatggcttat caaattcaga aaactcgttt cagcctgtac
4981 ccttaatctg acctaggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
5041 ctctaaacgg gtcttgaggg gttttttgct gaaacctcag gcatttgaga agcacacggt
5101 cacactgctt ccggtagtca ataaaccggt aaaccagcaa tagacataag cggctattta
5161 acgaccctgc cctgaaccga cgaccgggtc atcgtggccg gatcttgcgg cccctcggct
5221 tgaacgaatt gttagacatt atttgccgac taccttggtg atctcgcctt tcacgtagtg
5281 gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt gtccaagata
5341 agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca
5401 gtcggcagcg acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga
5461 caacgtaagc actacatttc gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt
5521 taaggtttca tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc
5581 cgccgctgga cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag
5641 atcaatgtcg atcgtggctg gctcgaagat acctgcaaga atgtcattgc gctgccattc
5701 tccaaattgc agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac
5761 aatggtgact tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa
5821 aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag
5881 caaatcaata tcactgtgtg gcttcaggcc gccatccact gcggagccgt acaaatgtac
5941 ggccagcaac gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt
6001 cgatacttcg gcgatcaccg cttccctcat actcttcctt tttcaatatt attgaagcat
6061 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
6121 aatagctagc tcactcggtc gctacgctcc gggcgtgaga ctgcggcggg cgctgcggac
6181 acatacaaag ttacccacag attccgtgga taagcagggg actaacatgt gaggcaaaac
6241 agcagggccg cgccggtggc gtttttccat aggctccgcc ctcctgccag agttcacata
6301 aacagacgct tttccggtgc atctgtggga gccgtgaggc tcaaccatga atctgacagt
6361 acgggcgaaa cccgacagga cttaaagatc cccaccgttt ccggcgggtc gctccctctt
6421 gcgctctcct gttccgaccc tgccgtttac cggatacctg ttccgccttt ctcccttacg
6481 ggaagtgtgg cgctttctca tagctcacac actggtatct cggctcggtg taggtcgttc
6541 gctccaagct gggctgtaag caagaactcc ccgttcagcc cgactgctgc gccttatccg
6601 gtaactgttc acttgagtcc aacccggaaa agcacggtaa aacgccactg gcagcagcca
6661 ttggtaactg ggagttcgca gaggatttgt ttagctaaac acgcggttgc tcttgaagtg
6721 tgcgccaaag tccggctaca ctggaaggac agatttggtt gctgtgctct gcgaaagcca
6781 gttaccacgg ttaagcagtt ccccaactga cttaaccttc gatcaaacca cctccccagg
6841 tggttttttc gaaggttaag tcagttgggg aactgcttaa ccttcgatca aaccacctcc
6901 ccaggtggtt ttttcgttta cagggcaaaa gattacgcgc agaaaaaaag gatctcaaga
6961 agatcctttg atcttttcta ctgaaccgct ctagatttca gtgcaattta tctcttcaaa
7021 tgtagcacct gaagtcagcc ccatacgata taagttgtaa ttctcatgtt agtcatgccc
7081 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
7141 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
7201 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
7261 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
7321 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
7381 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
7441 atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc
7501 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
7561 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
7621 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
7681 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
7741 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
7801 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
7861 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
7921 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
7981 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
8041 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
8101 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
8161 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
8221 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
8281 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
8341 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaaatta atacgactca
8401 ctatagggga attgtgagcg gataacaatt cccctgtaga aataattttg tttaacttta
8461 ataaggagat ataccatggg tgaactgccg agtaggtagc tgataacttg
//
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