质粒名称: | Ube1/PET21d ,pET21d-Ube1 |
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出品公司: | 爱迪基因 |
目录编号: | 34965 |
存储实验室: | Cynthia Wolberger |
载体骨架基本信息 |
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载体名称: | PET21d |
载体出品公司: | Novagen |
载体抗性: | 氨苄青霉素 |
空载体大小: | 5400 bp |
完整质粒大小: | -- |
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 |
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插入基因名称: | Ube1,UBA1 |
物种来源: | H. sapiens (human) |
插入大小: | 3176 bp |
基因突变: | -- |
GenBank编号 | NM_153280 |
启动子 | T7 |
标签/融合蛋白: | His(主干上的N终端) |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | BglII(未破坏) |
3' 克隆位点: | Not I (未破坏) |
5' 测序引物 | T7 |
3' 测序引物 | T7term |
文献引用方法 |
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材料和方法部分: | Ube1/PET21d was a gift from Cynthia Wolberger (Addgene plasmid # 34965 ; http://n2t.net/addgene:34965 ; RRID:Addgene_34965) |
参考文献部分: | A spectrophotometric assay for conjugation of ubiquitin and ubiquitin-like proteins. Berndsen CE, Wolberger C. Anal Biochem. 2011 Nov 1;418(1):102-10. Epub 2011 Jul 2. 10.1016/j.ab.2011.06.034 PubMed 21771579 |
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Ube1PET21d载体基本信息
Ube1PET21d载体图谱质粒图谱和多克隆位点信息
Ube1PET21d载体简介
Ube1PET21d载体序列
LOCUS Ube1/PET21d 8609 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Ube1/PET21d SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8609) AUTHORS Berndsen CE, Wolberger C TITLE A spectrophotometric assay for conjugation of ubiquitin and ubiquitin-like proteins. JOURNAL Anal Biochem. 2011 Nov 1;418(1):102-10. Epub 2011 Jul 2. PUBMED 21771579 REFERENCE 2 (bases 1 to 8609) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8609 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 106..125 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 209..228 /label=T7 /note="T7 promoter, forward primer" promoter 209..227 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 228..252 /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)." RBS 267..289 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 302..319 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 3549..3566 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(3619..3637) /label=T7 Term /note="T7 terminator, reverse primer" terminator 3633..3680 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 3717..4172 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3804..3823) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4014..4035 /label=F1ori-F /note="F1 origin, forward primer" promoter 4198..4302 /gene="bla" /label=AmpR promoter CDS 4303..5163 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(4521..4540) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5334..5922 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5823..5842 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6076..6093 /label=L4440 /note="L4440 vector, forward primer" misc_feature 6108..6247 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(6173..6192) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6333..6355 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(6349..6540) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(7349..8431) /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 8393..8412 /label=LacI-R /note="LacI, reverse primer" promoter complement(8432..8509) /gene="lacI" /label=lacI promoter ORIGIN 1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa 61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat 121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta 181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg 241 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat ataccatggg 301 tcatcaccat catcatcacg gaggtatgtc cagctcgccg ctgtccaaga aacgtcgcgt 361 gtccgggcct gatccaaagc cgggttctaa ctgctcccct gcccagtccg tgttgtccga 421 agtgccctcg gtgccaacca acggaatggc caagaacggc agtgaagcag acatagacga 481 gggcctttac tcccggcagc tgtatgtgtt gggccatgag gcaatgaagc ggctccagac 541 atccagtgtc ctggtatcag gcctgcgggg cctgggcgtg gagatcgcta agaacatcat 601 ccttggtggg gtcaaggctg ttaccctaca tgaccagggc actgcccagt gggctgatct 661 ttcctcccag ttctacctgc gggaggagga catcggtaaa aaccgggccg aggtatcaca 721 gccccgcctc gctgagctca acagctatgt gcctgtcact gcctacactg gacccctcgt 781 tgaggacttc cttagtggtt tccaggtggt ggtgctcacc aacacccccc tggaggacca 841 gctgcgagtg ggtgagttct gtcacaaccg tggcatcaag ctggtggtgg cagacacgcg 901 gggcctgttt gggcagctct tctgtgactt tggagaggaa atgatcctca cagattccaa 961 tggggagcag ccactcagtg ctatggtttc tatggttacc aaggacaacc ccggtgtggt 1021 tacctgcctg gatgaggccc gacacgggtt tgagagcggg gactttgtct ccttttcaga 1081 agtacagggc atggttgaac tcaacggaaa tcagcccatg gagatcaaag tcctgggtcc 1141 ttataccttt agcatctgtg acacctccaa cttctccgac tacatccgtg gaggcatcgt 1201 cagtcaggtc aaagtaccta agaagattag ctttaaatcc ttggtggcct cactggcaga 1261 acctgacttt gtggtgacgg acttcgccaa gttttctcgc cctgcccagc tgcacattgg 1321 cttccaggcc ctgcaccagt tctgtgctca gcatggccgg ccacctcggc cccgcaatga 1381 ggaggatgca gcagaactgg tagccttagc acaggctgtg aatgctcgag ccctgccagc 1441 agtgcagcaa aataacctgg acgaggacct catccggaag ctggcatatg tggctgctgg 1501 ggatctggca cccataaacg ccttcattgg gggcctggct gcccaggaag tcatgaaggc 1561 ctgctccggg aagttcatgc ccatcatgca gtggctatac tttgatgccc ttgagtgtct 1621 ccctgaggac aaagaggtcc tcacagagga caagtgcctc cagcgccaga accgttatga 1681 cgggcaagtg gctgtgtttg gctcagacct gcaagagaag ctgggcaagc agaagtattt 1741 cctggtgggt gcgggggcca ttggctgtga gctgctcaag aactttgcca tgattgggct 1801 gggctgcggg gagggtggag aaatcatcgt tacagacatg gacaccattg agaagtcaaa 1861 tctgaatcga cagtttcttt tccggccctg ggatgtcacg aagttaaagt ctgacacggc 1921 tgctgcagct gtgcgccaaa tgaatccaca tatccgggtg acaagccacc agaaccgtgt 1981 gggtcctgac acggagcgca tctatgatga cgattttttc caaaacctag atggcgtggc 2041 caatgccctg gacaacgtgg atgcccgcat gtacatggac cgccgctgtg tctactaccg 2101 gaagccactg ctggagtcag gcacactggg caccaaaggc aatgtgcagg tggtgatccc 2161 cttcctgaca gagtcgtaca gttccagcca ggacccacct gagaagtcca tccccatctg 2221 taccctgaag aacttcccta atgccatcga gcacaccctg cagtgggctc gggatgagtt 2281 tgaaggcctc ttcaagcagc cagcagaaaa tgtcaaccag tacctcacag accccaagtt 2341 tgtggagcga acactgcggc tggcaggcac tcagcccttg gaggtgctgg aggctgtgca 2401 gcgcagcctg gtgctgcagc gaccacagac ctgggctgac tgcgtgacct gggcctgcca 2461 ccactggcac acccagtact cgaacaacat ccggcagctg ctgcacaact tccctcctga 2521 ccagctcaca agctcaggag cgccgttctg gtctgggccc aaacgctgtc cacacccgct 2581 cacctttgat gtcaacaatc ccctgcatct ggactatgtg atggctgctg ccaacctgtt 2641 tgcccagacc tacgggctga caggctctca ggaccgagct gctgtggcca cattcctgca 2701 gtctgtgcag gtccccgaat tcacccccaa gtctggcgtc aagatccatg tttctgacca 2761 ggagctgcag agcgccaatg cctctgttga tgacagtcgt ctagaggagc tcaaagccac 2821 tctgcccagc ccagacaagc tccctggatt caagatgtac cccattgact ttgagaagga 2881 tgatgacagc aactttcata tggatttcat cgtggctgca tccaacctcc gggcagaaaa 2941 ctatgacatt ccttctgcag accggcacaa gagcaagctg attgcaggga agatcatccc 3001 agccattgcc acgaccacag cagccgtggt tggccttgtg tgtctggagc tgtacaaggt 3061 tgtgcagggg caccgacagc ttgactccta caagaatggt ttcctcaact tggccctgcc 3121 tttctttggt ttctctgaac cccttgccgc accacgtcac cagtactata accaagagtg 3181 gacattgtgg gatcgctttg aggtacaagg gctgcagcct aatggtgagg agatgaccct 3241 caaacagttc ctcgactatt ttaagacaga gcacaaatta gagatcacca tgctgtccca 3301 gggcgtgtcc atgctctatt ccttcttcat gccagctgcc aagctcaagg aacggttgga 3361 tcagccgatg acagagattg tgagccgtgt gtcgaagcga aagctgggcc gccacgtgcg 3421 ggcgctggtg cttgagctgt gctgtaacga cgagagcggc gaggatgtcg aggttcccta 3481 tgtccgatac accatccgct gaggatccga attcgagctc cgtcgacaag cttgcggccg 3541 cactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag 3601 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac 3661 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc 3721 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 3781 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 3841 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 3901 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc 3961 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 4021 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg 4081 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 4141 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc 4201 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa 4261 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc 4321 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa 4381 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa 4441 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg 4501 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa 4561 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc 4621 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc 4681 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta 4741 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag 4801 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca 4861 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata 4921 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc 4981 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca 5041 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca 5101 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg 5161 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa 5221 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt 5281 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat 5341 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 5401 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga 5461 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac 5521 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt 5581 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag 5641 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc 5701 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag 5761 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca 5821 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt 5881 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc 5941 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc 6001 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc 6061 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat 6121 tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct 6181 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat 6241 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc 6301 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc 6361 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag 6421 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt 6481 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac 6541 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga 6601 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga 6661 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg 6721 ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat 6781 gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac 6841 acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc 6901 gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca 6961 gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg ccgccatgcc 7021 ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg 7081 agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca 7141 gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg 7201 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa 7261 ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt 7321 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 7381 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 7441 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct 7501 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct 7561 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca 7621 ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca 7681 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt 7741 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct 7801 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag 7861 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca 7921 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag 7981 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct 8041 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca 8101 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac 8161 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca 8221 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc 8281 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag 8341 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact 8401 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg 8461 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga 8521 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg 8581 agcaccgccg ccgcaaggaa tggtgcatg //
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Ube1PET21d载体质粒应用举例
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