| 质粒名称: | pWUR702 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 53079 |
| 存储实验室: | John van der Oost |
|
载体骨架基本信息 |
|
| 载体名称: | pCDF-1b |
| 载体出品公司: | EMD Millipore |
| 载体抗性: | 大观霉素 |
| 空载体大小: | 3621 bp |
| 完整质粒大小: | 5844 bp |
| 载体类型: | 细菌表达 |
| 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: | TtAgo |
| 物种来源: | Thermus thermophilus |
| 插入大小: | 2223 bp |
| 基因又名: | TTHB068 |
| GenBank编号 | AP008227 |
| 启动子 | T7 |
| 标签/融合蛋白: | Strep-II tag (N terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NcoI(未破坏) |
| 3' 克隆位点: | AvrII (未破坏) |
| 5' 测序引物 | T7 |
| 3' 测序引物 | T7-Term |
|
文献引用方法 |
|
| 材料和方法部分: | pWUR702 was a gift from John van der Oost (爱迪基因 plasmid # 53079) |
| 参考文献部分: | DNA-guided DNA interference by a prokaryotic Argonaute. Swarts DC, Jore MM, Westra ER, Zhu Y, Janssen JH, Snijders AP, Wang Y, Patel DJ, Berenguer J, Brouns SJ, van der Oost J. Nature. 2014 Mar 13;507(7491):258-61. doi: 10.1038/nature12971. Epub 2014 Feb 16. 10.1038/nature12971 PubMed 24531762 |
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 pWUR702 5634 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Inducible TtAgo expression.
ACCESSION .
VERSION .
KEYWORDS pWUR702
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5634)
AUTHORS Swarts DC, Jore MM, Westra ER, Zhu Y, Janssen JH, Snijders AP, Wang
Y, Patel DJ, Berenguer J, Brouns SJ, van der Oost J
TITLE DNA-guided DNA interference by a prokaryotic Argonaute.
JOURNAL Nature. 2014 Mar 13;507(7491):258-61. doi: 10.1038/nature12971. Epub
2014 Feb 16.
PUBMED 24531762
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pWUR702|
FEATURES Location/Qualifiers
source 1..5634
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 3..27
/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)."
CDS 80..103
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 110..133
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
CDS 2225..2242
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS 2249..2278
/codon_start=1
/product="10xHis affinity tag"
/label=10xHis
/translation="HHHHHHHHHH"
primer_bind complement(2301..2319)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2315..2362
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(2533..3324)
/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(3325..3416)
/gene="bla"
/label=AmpR promoter
rep_origin complement(3464..4202)
/direction=LEFT
/label=CloDF13 ori
/note="Plasmids containing the CloDF13 (CDF) origin of
replication can be propagated in E. coli cells that contain
additional plasmids with compatible origins."
CDS complement(4412..5494)
/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 5456..5475
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(5495..5572)
/gene="lacI"
/label=lacI promoter
primer_bind 5618..3
/label=T7
/note="T7 promoter, forward primer"
promoter 5618..2
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
61 gagatatacc atgtcaagct ggagccaccc gcagttcgaa aagggtgcac ttgaagtcct
121 ctttcaggga cccgggtacc agatgaacca ccttggaaaa acggaagtgt tcctaaaccg
181 cttcgcccta cggcctctga accccgagga actcaggcct tggcgcctcg aggtggttct
241 ggaccctcct ccggggcggg aggaggtgta tccccttctc gcccaggtgg cccgccgggc
301 ggggggcgtc acggtgcgca tgggagacgg cctcgcctcc tggtcacccc ctgaggtcct
361 ggtcttggag ggcaccttgg cgcggatggg gcagaactac gcctaccgcc tctaccccaa
421 ggggaggagg cctctggacc ccaaggaccc gggggagcgg agcgtgcttt cggccctagc
481 ccgaaggctc cttcaggagc gcctcaggcg cctcgagggg gtctgggtgg aggggcttgc
541 ggtgtaccgg agggagcacg cccgggggcc cgggtggcgg gtgcttgggg gggcggtctt
601 ggacctttgg gtctccgact cgggggcgtt cctcctggag gtggaccccg cttaccgcat
661 cctctgcgag atgtccctcg aggcctggct tgcccaaggc caccctctgc ccaaacgggt
721 ccggaacgcc tacgaccggc gcacctggga gcttctccgg ctgggagagg aagaccccaa
781 ggagctccct ctcccgggcg gcttgagcct cctggactac cacgctttca agggccgcct
841 ccagggccgg gaaggggggc gggtggcctg ggtggcggac ccaaaggatc cgcgaaagcc
901 aatcccccat ctgacgggcc ttttggtgcc cgttctgacc ctggaagacc ttcatgaaga
961 ggagggaagc ctggccctct ccttgccttg ggaagagcgg cgtcggcgga cccgggagat
1021 cgccagctgg atcggccggc gcctggggct gggcacacct gaggcggtgc gcgcccaggc
1081 ttaccgcttg agcatcccca agctcatggg cagaagagcc gtgagcaagc ctgcggacgc
1141 cctccgcgtg gggttttacc gggcccagga aaccgccctg gccctcttgc gcctggacgg
1201 ggcccaagga tggccggagt ttctccggcg ggccttgctc cgggcctttg gggcgagcgg
1261 ggcttccctc cgtttgcaca ccctccacgc ccatccctcc caaggcctgg ctttccgcga
1321 ggcgctgcgg aaggccaagg aggagggggt ccaggccgtg ctggtcctca ccccgcccat
1381 ggcctgggaa gaccgcaacc gcctgaaggc ccttcttctc agggaaggcc ttcccagcca
1441 aatcctcaac gtccccctcc gggaggagga acgccaccgc tgggagaacg ccctcctggg
1501 cctcctggcc aaagcggggc ttcaggtggt ggccctgagc ggcgcctatc cggcggagct
1561 cgccgtgggc tttgacgccg gcggaaggga gtcctttcgc ttcgggggcg cggcctgcgc
1621 cgtgggcggg gacggcggcc acctcctctg gaccctcccc gaggcccagg ccggggagcg
1681 gatcccccag gaggtggtct gggacctctt ggaggagacc ctctgggcct tcagacgcaa
1741 ggcggggagg cttccttccc gggtcctcct ccttcgggac ggccgcgtgc cccaggacga
1801 gttcgccctg gccttggagg cccttgcccg ggaaggcata gcctacgact tggtttcggt
1861 gcgcaagtcg ggtggggggc gggtctaccc cgtgcagggg cgcctggcgg acgggcttta
1921 cgtccccttg gaggacaaga cctttcttct ccttaccgtc caccgggact tccggggcac
1981 gccccgaccc ctgaagctgg tgcacgaggc gggggacacg cccctcgagg ccctggccca
2041 ccagattttc catctgaccc gcctctaccc ggcgagcggt ttcgccttcc cccggcttcc
2101 cgctcccctt cacctggccg accgcctggt gaaggaggtg ggccggttgg ggatccgtca
2161 cctcaaggag gtggaccggg aaaagctctt cttcgtttag gaattcgcgg ccgcagagct
2221 cgctctggtg ccacgcggta gttccgctca tcaccaccat catcaccatc accaccacta
2281 attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
2341 acgggtcttg aggggttttt tgctgaaacc tcaggcattt gagaagcaca cggtcacact
2401 gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc
2461 ctgccctgaa ccgacgaccg ggtcatcgtg gccggatctt gcggcccctc ggcttgaacg
2521 aattgttaga cattatttgc cgactacctt ggtgatctcg cctttcacgt agtggacaaa
2581 ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa gataagcctg
2641 tctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc
2701 agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt
2761 aagcactaca tttcgctcat cgccagccca gtcgggcggc gagttccata gcgttaaggt
2821 ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc
2881 tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat
2941 gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa
3001 ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt
3061 gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc
3121 gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc
3181 aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag
3241 caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac
3301 ttcggcgatc accgcttccc tcatactctt cctttttcaa tattattgaa gcatttatca
3361 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagc
3421 tagctcactc ggtcgctacg ctccgggcgt gagactgcgg cgggcgctgc ggacacatac
3481 aaagttaccc acagattccg tggataagca ggggactaac atgtgaggca aaacagcagg
3541 gccgcgccgg tggcgttttt ccataggctc cgccctcctg ccagagttca cataaacaga
3601 cgcttttccg gtgcatctgt gggagccgtg aggctcaacc atgaatctga cagtacgggc
3661 gaaacccgac aggacttaaa gatccccacc gtttccggcg ggtcgctccc tcttgcgctc
3721 tcctgttccg accctgccgt ttaccggata cctgttccgc ctttctccct tacgggaagt
3781 gtggcgcttt ctcatagctc acacactggt atctcggctc ggtgtaggtc gttcgctcca
3841 agctgggctg taagcaagaa ctccccgttc agcccgactg ctgcgcctta tccggtaact
3901 gttcacttga gtccaacccg gaaaagcacg gtaaaacgcc actggcagca gccattggta
3961 actgggagtt cgcagaggat ttgtttagct aaacacgcgg ttgctcttga agtgtgcgcc
4021 aaagtccggc tacactggaa ggacagattt ggttgctgtg ctctgcgaaa gccagttacc
4081 acggttaagc agttccccaa ctgacttaac cttcgatcaa accacctccc caggtggttt
4141 tttcgtttac agggcaaaag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
4201 tcttttctac tgaaccgctc tagatttcag tgcaatttat ctcttcaaat gtagcacctg
4261 aagtcagccc catacgatat aagttgtaat tctcatgtta gtcatgcccc gcgcccaccg
4321 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
4381 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
4441 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
4501 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
4561 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
4621 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
4681 ccactaccga gatgtccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
4741 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
4801 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
4861 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
4921 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
4981 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
5041 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
5101 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
5161 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
5221 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
5281 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
5341 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
5401 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
5461 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
5521 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct
5581 cgacgctctc ccttatgcga ctcctgcatt aggaaattaa tacgactcac tata
//
