| 质粒名称: | pQCascade |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 140622 |
| 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: | 8493 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, CRISPR(entry) |
| 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 was a gift from Sheng Yang (Addgene plasmid # 140622 ; http://n2t.net/addgene:140622 ; RRID:Addgene_140622) |
| 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 8493 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses V. cholerae TniQ, Cas8, Cas7, and Cas6 and CRISPR RNA.
ACCESSION .
VERSION .
KEYWORDS pQCascade
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8493)
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 8493)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8493
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 127..145
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 146..170
/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(4968..4986)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 4982..5029
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(5200..5991)
/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(5992..6083)
/gene="bla"
/label=AmpR promoter
CDS complement(7144..8226)
/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 8188..8207
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(8227..8304)
/gene="lacI"
/label=lacI promoter
promoter 8350..8368
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 8369..8393
/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 gagtaggtag ctgataacgg atccgaattc gagctcggcg cgcctgcagg tcgacaagct
61 tgcggccgca taatgcttaa gtcgaacaga aagtaatcgt attgtacacg gccgcataat
121 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc ccatcttagt
181 atattagtta agtataagaa ggagatatac atatgttttt gcaaagacct aaaccttaca
241 gcgatgaaag tttagaaagt ttctttatcc gagtggctaa caaaaatggc tacggtgatg
301 tccatcgctt cctagaagcc actaaacgat tccttcaaga cattgaccat aatggctatc
361 aaacctttcc gactgatata actcggataa acccatactc agctaaaaac agttccagcg
421 cacgaactgc gtcattcctg aaacttgcac aattgacatt taatgaaccg ccagagctac
481 ttgggttggc aattaacaga acaaacatga aatactcgcc gtcaactagc gcggttgtta
541 gaggtgcaga agtctttcct cgcagtttac tacggacgca ctccatcccc tgctgtcctt
601 tgtgtctgcg agaaaatggc tacgcctcct acctttggca ctttcagggg tacgaatact
661 gccacagcca taacgtacct ttaattacca cttgtagctg tggtaaggag tttgactacc
721 gagtatctgg gttaaagggc atttgctgca aatgcaagga gcctatcacc ttaaccagca
781 gggagaacgg tcatgaggca gcgtgtactg tttcaaactg gcttgctggc catgaatcta
841 aacctctgcc aaatcttcct aaaagctacc gatggggttt agttcattgg tggatgggta
901 ttaaagatag cgagttcgat cacttttcgt tcgttcaatt tttctcaaac tggccaaggt
961 cattccactc gataatcgaa gatgaagtag agttcaacct tgagcatgct gttgtcagca
1021 cgtctgaatt acgactaaaa gatcttcttg gtcgattgtt tttcggttca attcggttac
1081 ctgagcggaa tcttcaacac aatatcatcc ttggtgagct tctctgctat ttagaaaatc
1141 gtttatggca agacaaggga ttaatcgcca acctcaaaat gaacgcgtta gaggcgactg
1201 taatgttaaa ttgtagcctc gatcagattg catcaatggt tgaacaacgc atcttgaagc
1261 caaatcgaaa aagcaagccc aacagccctc ttgatgttac cgattatcta tttcatttcg
1321 gcgatatttt ctgtctttgg ttagctgagt tccaaagcga tgagtttaac cgttcgtttt
1381 atgtgtcgag gtggtaaatg caaactctga aagaactaat cgcatccaat cctgacgact
1441 taacaactga acttaagaga gcatttcgtc cactcacacc gcatattgca attgatggta
1501 atgaacttga cgcactgacg atattagtca atttaaccga taagactgat gatcagaaag
1561 acctgctcga tcgagccaaa tgcaagcaaa aacttcgaga tgaaaaatgg tgggctagct
1621 gcataaattg cgttaattac agacaaagcc ataacccaaa attcccggat atacgttctg
1681 aaggcgtgat tcgaacccaa gccctgggtg aattaccgag tttcctactc tcgtcctcaa
1741 aaatcccacc ataccattgg tcatatagcc atgattcaaa atacgtcaac aaaagcgcat
1801 tcctcaccaa tgagttttgt tgggatggtg agatctcatg tttgggtgag cttcttaaag
1861 atgcagatca cccactttgg aatactttga aaaagttagg ttgttctcaa aaaacttgca
1921 aagcaatggc aaaacaacta gctgatatta ctctcacgac tatcaatgtc acactcgcac
1981 caaattacct gactcaaata tctctccccg atagtgatac atcctacatt tcactctctc
2041 ctgttgcatc gctatcgatg caaagccact ttcatcagag gctccaagat gaaaatcggc
2101 atagcgcgat aacgcggttt agccgaacta ccaacatggg ggtcactgcg atgacatgtg
2161 gcggcgcatt tagaatgtta aagtctggcg ctaagttttc cagcccccct catcaccgat
2221 taaacagtaa acgaagttgg ttgacgtcag agcatgttca gtcattaaaa cagtaccagc
2281 gcctcaacaa aagcctcata cctgaaaact ctcggattgc actccgtaga aaatacaaaa
2341 tcgagcttca aaatatggtc agatcttggt ttgcaatgca agaccatacc cttgattcga
2401 acatacttat ccaacatttg aatcatgacc tatcttactt aggagccaca aaacgttttg
2461 catacgatcc agcgatgacc aagctcttta ctgagctttt gaaacgagag ttatcaaatt
2521 caatcaataa tggtgagcaa cacactaatg gatcgttttt agtcctaccg aatatcaggg
2581 tttgtggcgc aacagcttta agctccccgg taacggtggg gattccatca cttacagctt
2641 tctttggctt cgttcacgca tttgaacgga atataaatcg caccacctca tcgtttcgtg
2701 ttgaatcctt tgcgatatgc gtccatcaac tacatgtcga gaagcgaggt ttgacagcag
2761 agtttgtgga aaaaggcgac gggactatat ccgctcccgc gacccgggat gactggcagt
2821 gtgatgtcgt atttagcctt attttgaaca ccaactttgc tcaacatatt gaccaagata
2881 cgttagttac atcactacca aagcgattgg ctcggggttc agcaaaaatt gcgattgatg
2941 actttaaaca tatcaactca ttctcgacat tagaaacagc gatcgaatct ctgccaatag
3001 aagctggtag gtggttatca ctttacgcac agtcaaacaa taatctaagt gatctattag
3061 cagccatgac agaggaccat cagctcatgg caagctgcgt cggttaccac ttgttagaag
3121 agcccaaaga taaaccaaac tccctcagag gttacaaaca cgctatcgcc gagtgcatca
3181 ttggactcat taactcaatc acctttagct cagagactga tcccaacaca atcttttggt
3241 cgctaaagaa ctatcaaaac tacctagtgg tacagccaag gagtatcaac gatgaaacta
3301 ccgacaaatc tagcctatga gcgctctatc gacccatcag atgtctgttt ttttgtcgtc
3361 tggcccgatg atagaaaaac acctttaacc tacaattctc gtactctgct cgggcaaatg
3421 gaagcggcat cattagccta tgatgtctca ggtcaaccaa taaaaagtgc caccgctgag
3481 gcgttagctc aagggaaccc tcatcaagtt gatttctgcc acgttccata cggtgcgagt
3541 catattgaat gcagtttctc cgtctcgttt tcttctgaac tacgtcaacc atataagtgt
3601 aactcaagca aagttaaaca aacgctagtg caattagtcg agctctacga aacgaaaatc
3661 ggctggactg agctagcaac ccgatatttg atgaacattt gcaacggtaa atggctgtgg
3721 aaaaataccc gtaaagccta ttgctggaac attgtactta caccttggcc ttggaacggg
3781 gaaaaggttg gatttgaaga tattcgtact aactacacct cacggcaaga ctttaaaaat
3841 aataaaaatt ggtctgctat agttgaaatg atcaaaaccg cattttctag tactgatggg
3901 ctggcgatat ttgaagtcag ggccaccttg cacttgccaa cgaatgctat ggtgcggcca
3961 agccaagttt tcacagaaaa agaaagtggc agtaaaagta aatctaaaac tcaaaacagt
4021 cgagtttttc agagtacaac tattgatggt gaacgatcgc caatactagg ggcctttaaa
4081 acgggagcag ctattgcaac cattgacgac tggtatcctg aagccactga gccactaagg
4141 gtcggacggt ttggggttca tcgcgaagat gtcacttgct accgtcatcc gtctaccgga
4201 aaagattttt tctcgatatt acaacaagca gagcactata ttgaagtgtt gagcgccaac
4261 aaaactcccg ctcaagaaac tatcaacgac atgcactttt taatggctaa cctgattaag
4321 ggtgggatgt tccagcataa aggagactga ctgtgaaatg gtattataag acaatcacct
4381 ttctgccaga gttgtgcaac aacgagtcac tggctgcaaa gtgtctccgc gttctgcatg
4441 gatttaacta tcagtatgag acacgaaata tcggcgtttc atttccgctt tggtgtgatg
4501 caacggttgg aaaaaagatt tcatttgtca gcaagaacaa gatagaactc gacttactac
4561 ttaaacaaca ctatttcgtc caaatggaac aacttcaata ctttcatata tccaacactg
4621 ttctcgtccc agaagattgt acatacgttt cctttagacg ctgtcaatct atagataagc
4681 tcacagcagc agggctggca aggaaaatca gacgcctgga gaaacgtgct ctatctagag
4741 gcgagcaatt tgacccatca tcttttgctc aaaaagagca tactgcaata gcgcactacc
4801 actcacttgg ggagtccagc aaacagacga accgcaactt tcgactcaat atcaggatgc
4861 tctcggagca accccgtgag gggaactcga tttttagtag ctatggctta tcaaattcag
4921 aaaactcgtt tcagcctgta cccttaatct gacctaggct gctgccaccg ctgagcaata
4981 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaacctca
5041 ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca
5101 atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt catcgtggcc
5161 ggatcttgcg gcccctcggc ttgaacgaat tgttagacat tatttgccga ctaccttggt
5221 gatctcgcct ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg aggccaagcg
5281 atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc
5341 cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac
5401 tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc
5461 gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg
5521 aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc
5581 ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag
5641 aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg
5701 aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc
5761 aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag
5821 ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac
5881 tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc
5941 aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca tactcttcct
6001 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
6061 atgtatttag aaaaataaac aaatagctag ctcactcggt cgctacgctc cgggcgtgag
6121 actgcggcgg gcgctgcgga cacatacaaa gttacccaca gattccgtgg ataagcaggg
6181 gactaacatg tgaggcaaaa cagcagggcc gcgccggtgg cgtttttcca taggctccgc
6241 cctcctgcca gagttcacat aaacagacgc ttttccggtg catctgtggg agccgtgagg
6301 ctcaaccatg aatctgacag tacgggcgaa acccgacagg acttaaagat ccccaccgtt
6361 tccggcgggt cgctccctct tgcgctctcc tgttccgacc ctgccgttta ccggatacct
6421 gttccgcctt tctcccttac gggaagtgtg gcgctttctc atagctcaca cactggtatc
6481 tcggctcggt gtaggtcgtt cgctccaagc tgggctgtaa gcaagaactc cccgttcagc
6541 ccgactgctg cgccttatcc ggtaactgtt cacttgagtc caacccggaa aagcacggta
6601 aaacgccact ggcagcagcc attggtaact gggagttcgc agaggatttg tttagctaaa
6661 cacgcggttg ctcttgaagt gtgcgccaaa gtccggctac actggaagga cagatttggt
6721 tgctgtgctc tgcgaaagcc agttaccacg gttaagcagt tccccaactg acttaacctt
6781 cgatcaaacc acctccccag gtggtttttt cgaaggttaa gtcagttggg gaactgctta
6841 accttcgatc aaaccacctc cccaggtggt tttttcgttt acagggcaaa agattacgcg
6901 cagaaaaaaa ggatctcaag aagatccttt gatcttttct actgaaccgc tctagatttc
6961 agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta
7021 attctcatgt tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
7081 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
7141 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
7201 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
7261 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
7321 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
7381 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg
7441 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
7501 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
7561 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
7621 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
7681 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
7741 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
7801 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
7861 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
7921 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
7981 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
8041 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
8101 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
8161 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
8221 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
8281 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
8341 ttaggaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctgtag
8401 aaataatttt gtttaacttt aataaggaga tataccatgg gtgaactgcc gagtaggtag
8461 ctgataacga gacctctggt ctcgtgaact gcc
//
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