| 质粒名称: | pQCascade-IS1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 140624 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pCDFDuet-1 |
| 载体出品公司: | -- |
| 载体抗性: | 大观霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达，CRISPR；转座子 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | VchTniQ, VchCas8, VchCas7, VchCas6, crRNA-IS1 |
| 物种来源: | Vibrio cholera |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pQCascade-IS1 was a gift from Sheng Yang (Addgene plasmid # 140624 ; http://n2t.net/addgene:140624 ; RRID:Addgene_140624) |
| 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
pQCascadeIS1载体基本信息
pQCascadeIS1载体图谱质粒图谱和多克隆位点信息
pQCascadeIS1载体简介
pQCascadeIS1载体序列
LOCUS pQCascade-IS1 8510 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses V. cholerae TniQ, Cas8, Cas7, and Cas6 and crRNA-IS1. The
crRNA-IS1 targets IS1 loci in Escherchia coli..
ACCESSION .
VERSION .
KEYWORDS pQCascade-IS1
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.snapgene.com
FEATURES Location/Qualifiers
source 1..8510
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 147..165
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 166..190
/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(4988..5006)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 5002..5049
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(5220..6011)
/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(6012..6103)
/gene="bla"
/label=AmpR promoter
CDS complement(7164..8246)
/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 8208..8227
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(8247..8324)
/gene="lacI"
/label=lacI promoter
promoter 8370..8388
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 8389..8413
/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 gctgtcgttc gtgaactgcc gagtaggtag ctgataacgg atccgaattc gagctcggcg
61 cgcctgcagg tcgacaagct tgcggccgca taatgcttaa gtcgaacaga aagtaatcgt
121 attgtacacg gccgcataat cgaaattaat acgactcact ataggggaat tgtgagcgga
181 taacaattcc ccatcttagt atattagtta agtataagaa ggagatatac atatgttttt
241 gcaaagacct aaaccttaca gcgatgaaag tttagaaagt ttctttatcc gagtggctaa
301 caaaaatggc tacggtgatg tccatcgctt cctagaagcc actaaacgat tccttcaaga
361 cattgaccat aatggctatc aaacctttcc gactgatata actcggataa acccatactc
421 agctaaaaac agttccagcg cacgaactgc gtcattcctg aaacttgcac aattgacatt
481 taatgaaccg ccagagctac ttgggttggc aattaacaga acaaacatga aatactcgcc
541 gtcaactagc gcggttgtta gaggtgcaga agtctttcct cgcagtttac tacggacgca
601 ctccatcccc tgctgtcctt tgtgtctgcg agaaaatggc tacgcctcct acctttggca
661 ctttcagggg tacgaatact gccacagcca taacgtacct ttaattacca cttgtagctg
721 tggtaaggag tttgactacc gagtatctgg gttaaagggc atttgctgca aatgcaagga
781 gcctatcacc ttaaccagca gggagaacgg tcatgaggca gcgtgtactg tttcaaactg
841 gcttgctggc catgaatcta aacctctgcc aaatcttcct aaaagctacc gatggggttt
901 agttcattgg tggatgggta ttaaagatag cgagttcgat cacttttcgt tcgttcaatt
961 tttctcaaac tggccaaggt cattccactc gataatcgaa gatgaagtag agttcaacct
1021 tgagcatgct gttgtcagca cgtctgaatt acgactaaaa gatcttcttg gtcgattgtt
1081 tttcggttca attcggttac ctgagcggaa tcttcaacac aatatcatcc ttggtgagct
1141 tctctgctat ttagaaaatc gtttatggca agacaaggga ttaatcgcca acctcaaaat
1201 gaacgcgtta gaggcgactg taatgttaaa ttgtagcctc gatcagattg catcaatggt
1261 tgaacaacgc atcttgaagc caaatcgaaa aagcaagccc aacagccctc ttgatgttac
1321 cgattatcta tttcatttcg gcgatatttt ctgtctttgg ttagctgagt tccaaagcga
1381 tgagtttaac cgttcgtttt atgtgtcgag gtggtaaatg caaactctga aagaactaat
1441 cgcatccaat cctgacgact taacaactga acttaagaga gcatttcgtc cactcacacc
1501 gcatattgca attgatggta atgaacttga cgcactgacg atattagtca atttaaccga
1561 taagactgat gatcagaaag acctgctcga tcgagccaaa tgcaagcaaa aacttcgaga
1621 tgaaaaatgg tgggctagct gcataaattg cgttaattac agacaaagcc ataacccaaa
1681 attcccggat atacgttctg aaggcgtgat tcgaacccaa gccctgggtg aattaccgag
1741 tttcctactc tcgtcctcaa aaatcccacc ataccattgg tcatatagcc atgattcaaa
1801 atacgtcaac aaaagcgcat tcctcaccaa tgagttttgt tgggatggtg agatctcatg
1861 tttgggtgag cttcttaaag atgcagatca cccactttgg aatactttga aaaagttagg
1921 ttgttctcaa aaaacttgca aagcaatggc aaaacaacta gctgatatta ctctcacgac
1981 tatcaatgtc acactcgcac caaattacct gactcaaata tctctccccg atagtgatac
2041 atcctacatt tcactctctc ctgttgcatc gctatcgatg caaagccact ttcatcagag
2101 gctccaagat gaaaatcggc atagcgcgat aacgcggttt agccgaacta ccaacatggg
2161 ggtcactgcg atgacatgtg gcggcgcatt tagaatgtta aagtctggcg ctaagttttc
2221 cagcccccct catcaccgat taaacagtaa acgaagttgg ttgacgtcag agcatgttca
2281 gtcattaaaa cagtaccagc gcctcaacaa aagcctcata cctgaaaact ctcggattgc
2341 actccgtaga aaatacaaaa tcgagcttca aaatatggtc agatcttggt ttgcaatgca
2401 agaccatacc cttgattcga acatacttat ccaacatttg aatcatgacc tatcttactt
2461 aggagccaca aaacgttttg catacgatcc agcgatgacc aagctcttta ctgagctttt
2521 gaaacgagag ttatcaaatt caatcaataa tggtgagcaa cacactaatg gatcgttttt
2581 agtcctaccg aatatcaggg tttgtggcgc aacagcttta agctccccgg taacggtggg
2641 gattccatca cttacagctt tctttggctt cgttcacgca tttgaacgga atataaatcg
2701 caccacctca tcgtttcgtg ttgaatcctt tgcgatatgc gtccatcaac tacatgtcga
2761 gaagcgaggt ttgacagcag agtttgtgga aaaaggcgac gggactatat ccgctcccgc
2821 gacccgggat gactggcagt gtgatgtcgt atttagcctt attttgaaca ccaactttgc
2881 tcaacatatt gaccaagata cgttagttac atcactacca aagcgattgg ctcggggttc
2941 agcaaaaatt gcgattgatg actttaaaca tatcaactca ttctcgacat tagaaacagc
3001 gatcgaatct ctgccaatag aagctggtag gtggttatca ctttacgcac agtcaaacaa
3061 taatctaagt gatctattag cagccatgac agaggaccat cagctcatgg caagctgcgt
3121 cggttaccac ttgttagaag agcccaaaga taaaccaaac tccctcagag gttacaaaca
3181 cgctatcgcc gagtgcatca ttggactcat taactcaatc acctttagct cagagactga
3241 tcccaacaca atcttttggt cgctaaagaa ctatcaaaac tacctagtgg tacagccaag
3301 gagtatcaac gatgaaacta ccgacaaatc tagcctatga gcgctctatc gacccatcag
3361 atgtctgttt ttttgtcgtc tggcccgatg atagaaaaac acctttaacc tacaattctc
3421 gtactctgct cgggcaaatg gaagcggcat cattagccta tgatgtctca ggtcaaccaa
3481 taaaaagtgc caccgctgag gcgttagctc aagggaaccc tcatcaagtt gatttctgcc
3541 acgttccata cggtgcgagt catattgaat gcagtttctc cgtctcgttt tcttctgaac
3601 tacgtcaacc atataagtgt aactcaagca aagttaaaca aacgctagtg caattagtcg
3661 agctctacga aacgaaaatc ggctggactg agctagcaac ccgatatttg atgaacattt
3721 gcaacggtaa atggctgtgg aaaaataccc gtaaagccta ttgctggaac attgtactta
3781 caccttggcc ttggaacggg gaaaaggttg gatttgaaga tattcgtact aactacacct
3841 cacggcaaga ctttaaaaat aataaaaatt ggtctgctat agttgaaatg atcaaaaccg
3901 cattttctag tactgatggg ctggcgatat ttgaagtcag ggccaccttg cacttgccaa
3961 cgaatgctat ggtgcggcca agccaagttt tcacagaaaa agaaagtggc agtaaaagta
4021 aatctaaaac tcaaaacagt cgagtttttc agagtacaac tattgatggt gaacgatcgc
4081 caatactagg ggcctttaaa acgggagcag ctattgcaac cattgacgac tggtatcctg
4141 aagccactga gccactaagg gtcggacggt ttggggttca tcgcgaagat gtcacttgct
4201 accgtcatcc gtctaccgga aaagattttt tctcgatatt acaacaagca gagcactata
4261 ttgaagtgtt gagcgccaac aaaactcccg ctcaagaaac tatcaacgac atgcactttt
4321 taatggctaa cctgattaag ggtgggatgt tccagcataa aggagactga ctgtgaaatg
4381 gtattataag acaatcacct ttctgccaga gttgtgcaac aacgagtcac tggctgcaaa
4441 gtgtctccgc gttctgcatg gatttaacta tcagtatgag acacgaaata tcggcgtttc
4501 atttccgctt tggtgtgatg caacggttgg aaaaaagatt tcatttgtca gcaagaacaa
4561 gatagaactc gacttactac ttaaacaaca ctatttcgtc caaatggaac aacttcaata
4621 ctttcatata tccaacactg ttctcgtccc agaagattgt acatacgttt cctttagacg
4681 ctgtcaatct atagataagc tcacagcagc agggctggca aggaaaatca gacgcctgga
4741 gaaacgtgct ctatctagag gcgagcaatt tgacccatca tcttttgctc aaaaagagca
4801 tactgcaata gcgcactacc actcacttgg ggagtccagc aaacagacga accgcaactt
4861 tcgactcaat atcaggatgc tctcggagca accccgtgag gggaactcga tttttagtag
4921 ctatggctta tcaaattcag aaaactcgtt tcagcctgta cccttaatct gacctaggct
4981 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
5041 ggttttttgc tgaaacctca ggcatttgag aagcacacgg tcacactgct tccggtagtc
5101 aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg
5161 acgaccgggt catcgtggcc ggatcttgcg gcccctcggc ttgaacgaat tgttagacat
5221 tatttgccga ctaccttggt gatctcgcct ttcacgtagt ggacaaattc ttccaactga
5281 tctgcgcgcg aggccaagcg atcttcttct tgtccaagat aagcctgtct agcttcaagt
5341 atgacgggct gatactgggc cggcaggcgc tccattgccc agtcggcagc gacatccttc
5401 ggcgcgattt tgccggttac tgcgctgtac caaatgcggg acaacgtaag cactacattt
5461 cgctcatcgc cagcccagtc gggcggcgag ttccatagcg ttaaggtttc atttagcgcc
5521 tcaaatagat cctgttcagg aaccggatca aagagttcct ccgccgctgg acctaccaag
5581 gcaacgctat gttctcttgc ttttgtcagc aagatagcca gatcaatgtc gatcgtggct
5641 ggctcgaaga tacctgcaag aatgtcattg cgctgccatt ctccaaattg cagttcgcgc
5701 ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg
5761 cggagaatct cgctctctcc aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct
5821 cgccgcgttg tttcatcaag ccttacggtc accgtaacca gcaaatcaat atcactgtgt
5881 ggcttcaggc cgccatccac tgcggagccg tacaaatgta cggccagcaa cgtcggttcg
5941 agatggcgct cgatgacgcc aactacctct gatagttgag tcgatacttc ggcgatcacc
6001 gcttccctca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
6061 atgagcggat acatatttga atgtatttag aaaaataaac aaatagctag ctcactcggt
6121 cgctacgctc cgggcgtgag actgcggcgg gcgctgcgga cacatacaaa gttacccaca
6181 gattccgtgg ataagcaggg gactaacatg tgaggcaaaa cagcagggcc gcgccggtgg
6241 cgtttttcca taggctccgc cctcctgcca gagttcacat aaacagacgc ttttccggtg
6301 catctgtggg agccgtgagg ctcaaccatg aatctgacag tacgggcgaa acccgacagg
6361 acttaaagat ccccaccgtt tccggcgggt cgctccctct tgcgctctcc tgttccgacc
6421 ctgccgttta ccggatacct gttccgcctt tctcccttac gggaagtgtg gcgctttctc
6481 atagctcaca cactggtatc tcggctcggt gtaggtcgtt cgctccaagc tgggctgtaa
6541 gcaagaactc cccgttcagc ccgactgctg cgccttatcc ggtaactgtt cacttgagtc
6601 caacccggaa aagcacggta aaacgccact ggcagcagcc attggtaact gggagttcgc
6661 agaggatttg tttagctaaa cacgcggttg ctcttgaagt gtgcgccaaa gtccggctac
6721 actggaagga cagatttggt tgctgtgctc tgcgaaagcc agttaccacg gttaagcagt
6781 tccccaactg acttaacctt cgatcaaacc acctccccag gtggtttttt cgaaggttaa
6841 gtcagttggg gaactgctta accttcgatc aaaccacctc cccaggtggt tttttcgttt
6901 acagggcaaa agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
6961 actgaaccgc tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcagc
7021 cccatacgat ataagttgta attctcatgt tagtcatgcc ccgcgcccac cggaaggagc
7081 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
7141 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
7201 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
7261 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
7321 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
7381 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
7441 gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
7501 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
7561 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
7621 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
7681 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
7741 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
7801 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
7861 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
7921 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
7981 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
8041 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
8101 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
8161 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
8221 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
8281 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
8341 tcccttatgc gactcctgca ttaggaaatt aatacgactc actatagggg aattgtgagc
8401 ggataacaat tcccctgtag aaataatttt gtttaacttt aataaggaga tataccatgg
8461 gtgaactgcc gagtaggtag ctgataactg gcacggctgg gacggaagtc
//
