| 质粒名称: | pWUR790 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 101723 |
| 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: | Novagen |
| 载体抗性: | 链霉素 |
| 空载体大小: | 3621 bp |
| 完整质粒大小: | 5807 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | PfAgo |
| 物种来源: | Pyrococcus furiosus |
| 插入大小: | 2313 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | T7 Promotor |
| 标签/融合蛋白: | Strep(II)-tag（插入物上的N端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NcoI（销毁后未知） |
| 3' 克隆位点: | AvrII（销毁后未知） |
| 5' 测序引物 | TAATACGACTCACTATAGGG |
| 3' 测序引物 | GCTAGTTATTGCTCAGCGG |
|
文献引用方法 |
|
| 材料和方法部分: | pWUR790 was a gift from John van der Oost (Addgene plasmid # 101723 ; http://n2t.net/addgene:101723 ; RRID:Addgene_101723) |
| 参考文献部分: | Argonaute of the archaeon Pyrococcus furiosus is a DNA-guided nuclease that targets cognate DNA. Swarts DC, Hegge JW, Hinojo I, Shiimori M, Ellis MA, Dumrongkulraksa J, Terns RM, Terns MP, van der Oost J. Nucleic Acids Res. 2015 May 26;43(10):5120-9. doi: 10.1093/nar/gkv415. Epub 2015 Apr 29. 10.1093/nar/gkv415 PubMed 25925567 |
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 pWUR790 5872 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Synthetic codon optimized P. furiosus ago with N-terminal
Strep(II)-tag in pCDF-1b vector, expression vector for PfAgo.
ACCESSION .
VERSION .
KEYWORDS pWUR790
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5872)
AUTHORS Swarts DC, Hegge JW, Hinojo I, Shiimori M, Ellis MA, Dumrongkulraksa
J, Terns RM, Terns MP, van der Oost J
TITLE Argonaute of the archaeon Pyrococcus furiosus is a DNA-guided
nuclease that targets cognate DNA.
JOURNAL Nucleic Acids Res. 2015 May 26;43(10):5120-9. doi:
10.1093/nar/gkv415. Epub 2015 Apr 29.
PUBMED 25925567
REFERENCE 2 (bases 1 to 5872)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5872
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 1..20
/label=T7
/note="T7 promoter, forward primer"
promoter 1..19
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 20..44
/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 97..120
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 127..150
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
CDS 160..2472
/codon_start=1
/product="Argonaute protein from the archaeon Pyrococcus
furiosus"
/label=PfAgo
/note="DNA-guided nuclease (Swarts et al., 2015)"
/translation="MKAKVVINLVKINKKIIPDKIYVYRLFNDPEEELQKEGYSIYRLA
YENVGIVIDPENLIIATTKELEYEGEFIPEGEISFSELRNDYQSKLVLRLLKENGIGEY
ELSKLLRKFRKPKTFGDYKVIPSVEMSVIKHDEDFYLVIHIIHQIQSMKTLWELVNKDP
KELEEFLMTHKENLMLKDIASPLKTVYKPCFEEYTKKPKLDHNQEIVKYWYNYHIERYW
NTPEAKLEFYRKFGQVDLKQPAILAKFASKIKKNKNYKIYLLPQLVVPTYNAEQLESDV
AKEILEYTKLMPEERKELLENILAEVDSDIIDKSLSEIEVEKIAQELENKIRVRDDKGN
SVPISQLNVQKSQLLLWTNYSRKYPVILPYEVPEKFRKIREIPMFIILDSGLLADIQNF
ATNEFRELVKSMYYSLAKKYNSLAKKARSTNEIGLPFLDFRGKEKVITEDLNSDKGIIE
VVEQVSSFMKGKELGLAFIAARNKLSSEKFEEIKRRLFNLNVISQVVNEDTLKNKRDKY
DRNRLDLFVRHNLLFQVLSKLGVKYYVLDYRFNYDYIIGIDVAPMKRSEGYIGGSAVMF
DSQGYIRKIVPIKIGEQRGESVDMNEFFKEMVDKFKEFNIKLDNKKILLLRDGRITNNE
EEGLKYISEMFDIEVVTMDVIKNHPVRAFANMKMYFNLGGAIYLIPHKLKQAKGTPIPI
KLAKKRIIKNGKVEKQSITRQDVLDIFILTRLNYGSISADMRLPAPVHYAHKFANAIRN
EWKIKEEFLAEGFLYFV"
primer_bind complement(2491..2509)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2505..2552
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(2723..3514)
/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(3515..3606)
/gene="bla"
/label=AmpR promoter
CDS complement(4667..5749)
/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 5711..5730
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(5750..5827)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
61 tgtttaactt taataaggag atataccatg gcaagctggt cacatccgca gtttgaaaaa
121 ggtgcactgg aagttctgtt tcagggtcct gggtaccaga tgaaagcaaa agttgttatt
181 aacctggtga aaatcaacaa aaaaatcatc ccggataaaa tctacgtgta ccgcctgttt
241 aatgatccgg aagaagaact gcagaaagaa ggctatagca tttatcgtct ggcctatgaa
301 aatgtgggca ttgttatcga tccggaaaat ctgattattg ccaccaccaa agaactggaa
361 tatgaaggtg aatttattcc ggaaggcgaa atcagcttta gcgaactgcg taatgattat
421 cagagcaaac tggttctgcg tctgctgaaa gaaaatggta ttggtgaata tgaactgtcc
481 aaactgctgc gcaaatttcg taaaccgaaa acctttggcg attacaaagt tattccgagc
541 gttgaaatga gcgtgatcaa acacgatgaa gatttctatc tggtgatcca tatcatccat
601 cagatccaga gcatgaaaac cctgtgggaa ctggttaata aagatccgaa agagctggaa
661 gaatttctga tgacccataa agaaaacctg atgctgaaag atattgccag tccgctgaaa
721 accgtgtata aaccgtgttt tgaagaatat accaaaaaac cgaaactgga ccacaaccaa
781 gagatcgtga aatattggta taactatcac atcgagcgct attggaatac accggaagca
841 aaactggaat tctatcgcaa atttggtcag gtggatctga aacagcctgc aattctggca
901 aaatttgcaa gcaaaattaa aaaaaataaa aactacaaaa tctatctgct gccgcagctg
961 gttgttccga cctataatgc agaacagctg gaaagtgatg tggccaaaga aattctggaa
1021 tatacaaaac tgatgcctga ggaacgtaaa gagctgctgg aaaatattct ggcagaagtg
1081 gatagcgata tcatcgataa aagcctgagc gaaatcgagg ttgaaaaaat tgcacaagaa
1141 ctggaaaaca aaatccgcgt gcgtgatgat aaaggtaata gcgttccgat tagccagctg
1201 aatgttcaga aaagccagct gctgctgtgg accaattatt cacgtaaata tccggttatc
1261 ctgccgtatg aagtgccgga aaaatttcgc aaaattcgtg aaatcccgat gttcattatt
1321 ctggatagcg gtctgctggc agatattcag aactttgcaa ccaatgaatt tcgcgagctg
1381 gtcaaaagca tgtattatag cctggccaaa aaatacaact ccctggcaaa aaaagcacgc
1441 agcaccaatg aaattggtct gccgtttctg gattttcgcg gtaaagaaaa agtgatcacc
1501 gaagatctga atagcgataa aggcattatt gaagttgttg aacaggtgag cagctttatg
1561 aaaggtaaag aactgggtct ggcatttatt gcagcacgta ataaactgag cagcgagaaa
1621 tttgaagaaa tcaaacgccg tctgtttaac ctgaatgtta ttagccaggt ggtgaatgaa
1681 gataccctga aaaacaaacg cgataaatat gatcgtaatc gcctggacct gtttgttcgt
1741 cataatctgc tgttccaggt tctgagtaaa ctgggtgtta aatactatgt gctggactat
1801 cgcttcaact acgattatat cattggcatt gatgtggcac cgatgaaacg tagcgaaggt
1861 tatattggtg gtagcgcagt tatgtttgat agccagggtt atattcgtaa aatcgtgccg
1921 attaaaatcg gtgaacagcg tggtgaaagc gttgatatga acgaattttt caaagaaatg
1981 gtggacaaat tcaaagagtt caacatcaaa ctggataaca aaaaaatcct gctgctgcgt
2041 gatggtcgca ttaccaataa tgaagaagaa ggcctgaaat atatcagcga gatgttcgat
2101 attgaagtgg tgacaatgga tgtgatcaaa aaccatccgg ttcgtgcatt tgcaaacatg
2161 aaaatgtatt ttaacctggg tggtgccatt tatctgattc cgcataaact gaaacaggca
2221 aaaggcaccc cgattccgat taaactggcg aaaaaacgca ttatcaaaaa cggcaaagtg
2281 gaaaaacaga gcattacccg tcaggatgtt ctggatatct ttattctgac ccgtctgaat
2341 tatggtagca ttagcgcaga tatgcgtctg cctgcaccgg ttcattatgc acataaattt
2401 gccaatgcca ttcgcaacga gtggaaaatc aaagaagaat tcctggccga aggctttctg
2461 tattttgtgt aataacctag gctgctgcca ccgctgagca ataactagca taaccccttg
2521 gggcctctaa acgggtcttg aggggttttt tgctgaaacc tcaggcattt gagaagcaca
2581 cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta
2641 tttaacgacc ctgccctgaa ccgacgaccg ggtcatcgtg gccggatctt gcggcccctc
2701 ggcttgaacg aattgttaga cattatttgc cgactacctt ggtgatctcg cctttcacgt
2761 agtggacaaa ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa
2821 gataagcctg tctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg
2881 cccagtcggc agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc
2941 gggacaacgt aagcactaca tttcgctcat cgccagccca gtcgggcggc gagttccata
3001 gcgttaaggt ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt
3061 cctccgccgc tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag
3121 ccagatcaat gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc
3181 attctccaaa ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca
3241 caacaatggt gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt
3301 ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa
3361 ccagcaaatc aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat
3421 gtacggccag caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt
3481 gagtcgatac ttcggcgatc accgcttccc tcatactctt cctttttcaa tattattgaa
3541 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
3601 aacaaatagc tagctcactc ggtcgctacg ctccgggcgt gagactgcgg cgggcgctgc
3661 ggacacatac aaagttaccc acagattccg tggataagca ggggactaac atgtgaggca
3721 aaacagcagg gccgcgccgg tggcgttttt ccataggctc cgccctcctg ccagagttca
3781 cataaacaga cgcttttccg gtgcatctgt gggagccgtg aggctcaacc atgaatctga
3841 cagtacgggc gaaacccgac aggacttaaa gatccccacc gtttccggcg ggtcgctccc
3901 tcttgcgctc tcctgttccg accctgccgt ttaccggata cctgttccgc ctttctccct
3961 tacgggaagt gtggcgcttt ctcatagctc acacactggt atctcggctc ggtgtaggtc
4021 gttcgctcca agctgggctg taagcaagaa ctccccgttc agcccgactg ctgcgcctta
4081 tccggtaact gttcacttga gtccaacccg gaaaagcacg gtaaaacgcc actggcagca
4141 gccattggta actgggagtt cgcagaggat ttgtttagct aaacacgcgg ttgctcttga
4201 agtgtgcgcc aaagtccggc tacactggaa ggacagattt ggttgctgtg ctctgcgaaa
4261 gccagttacc acggttaagc agttccccaa ctgacttaac cttcgatcaa accacctccc
4321 caggtggttt tttcgaaggt taagtcagtt ggggaactgc ttaaccttcg atcaaaccac
4381 ctccccaggt ggttttttcg tttacagggc aaaagattac gcgcagaaaa aaaggatctc
4441 aagaagatcc tttgatcttt tctactgaac cgctctagat ttcagtgcaa tttatctctt
4501 caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca tgttagtcat
4561 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
4621 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
4681 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
4741 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
4801 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
4861 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
4921 cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa
4981 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
5041 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
5101 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
5161 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
5221 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
5281 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
5341 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
5401 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
5461 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
5521 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
5581 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
5641 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
5701 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
5761 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
5821 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa at
//
