| 质粒名称: | Ube1/PET21d ,pET21d-Ube1 |
|---|---|
| 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 |
|
| 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 |
|
| 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 |
|
| 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 |
买家导航
Ube1/PET21载体 Ube1PET21d质粒基本信息
Ube1/PET21质粒图谱 Ube1PET21d载体图谱和Ube1/PET21载体序列 Ube1PET21d质粒序列多克隆位点信息
Ube1/PET21质粒 Ube1PET21d载体简介
Ube1/PET21质粒序列 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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Ube1/PET21质粒 Ube1PET21d载体应用举例
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