| 质粒名称: | pEBB-BT-Ubiquitin |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 36098 |
| 存储实验室: | Ezra Burstein |
|
载体骨架基本信息 |
|
| 载体名称: | pEBB-TB |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5416 bp |
| 完整质粒大小: | 5894bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | Ubiquitin |
| 物种来源: | -- |
| 插入大小: | 228 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pEBB-BT-Ubiquitin was a gift from Ezra Burstein (Addgene plasmid # 36098 ; http://n2t.net/addgene:36098 ; RRID:Addgene_36098) |
| 参考文献部分: | A bimolecular affinity purification method under denaturing conditions for rapid isolation of a ubiquitinated protein for mass spectrometry analysis. Maine GN, Li H, Zaidi IW, Basrur V, Elenitoba-Johnson KS, Burstein E. Nat Protoc. 2010 Aug;5(8):1447-59. Epub 2010 Jul 22. 10.1038/nprot.2010.109 PubMed 20671728 |
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 pEBB-BT-Ubiquitin 5892 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pEBB-BT-Ubiquitin
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5892)
AUTHORS Maine GN, Li H, Zaidi IW, Basrur V, Elenitoba-Johnson KS, Burstein E
TITLE A bimolecular affinity purification method under denaturing
conditions for rapid isolation of a ubiquitinated protein for mass
spectrometry analysis.
JOURNAL Nat Protoc. 2010 Aug;5(8):1447-59. Epub 2010 Jul 22.
PUBMED 20671728
REFERENCE 2 (bases 1 to 5892)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5892
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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 186..208
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 205..221
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 252..462
/label=SV40 promoter
/note="SV40 early promoter"
promoter 274..470
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 321..456
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 383..402
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 558..1739
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 788..1730
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1687..1707
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
regulatory 1787..1796
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2048..2068
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 2114..2341
/codon_start=1
/product="human ubiquitin"
/label=ubiquitin
/translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
primer_bind complement(3026..3043)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3026..3042)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3035..3057)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3255..3710
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3342..3361)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3552..3573
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3727..3746)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3846..3868
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(3906..3924)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3992..4096
/gene="bla"
/label=AmpR promoter
CDS 4097..4957
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(4315..4334)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5128..5716
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5617..5636
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5870..5887
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttc
241 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
301 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
361 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
421 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctttgca
481 aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct tctaggtctt
541 gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
601 ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg
661 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
721 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
781 aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc
841 ttgcgtgcct tgaattactt ccacgcccct ggctgcagta cgtgattctt gatcccgagc
901 ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc
961 gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc
1021 ttcgcgcctg tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg
1081 ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg
1141 gtatttcggt ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt
1201 cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg
1261 gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa
1321 ggctggcccg gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg
1381 cagggagctc aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac
1441 aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg
1501 cgccgtccag gcacctcgat tagttctcga gcttttggag tacgtcgtct ttaggttggg
1561 gggaggggtt ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc
1621 cagcttggca cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt
1681 tcattctcaa gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtgag
1741 gattctctag agatccctcg acctcgagat ccattgtgct ggatctgcca ccatggctgg
1801 aaaggccggt gaaggtgaaa tccctgcccc tcttgctggt accgtttcta agatactggt
1861 aaaagaaggt gacactgtta aagctggtca aacagttctg gtgctggagg ctatgaaaat
1921 ggagacagaa attaacgctc ctactgacgg aaaagttgaa aaggtgttag ttaaggaaag
1981 agatgctgtt caaggtggtc aaggtctaat caagatcggc gactacgata tacccacaac
2041 cgccagcgag aatctgtact ttcaatcaga gttaaaaacc gccgcactag ctcaacatga
2101 cgaggctgga tccatgcaga ttttcgtgaa aacccttacg gggaagacca tcaccctcga
2161 ggttgaaccc tcggatacga tagaaaatgt aaaggccaag atccaggata aggaaggaat
2221 tcctcctgat cagcagagac tgatctttgc tggcaagcag ctggaagatg gacgtacttt
2281 gtctgactac aatattcaaa aggagtctac tcttcatctt gtgttgagac ttcgtggtgg
2341 ttaagcggcc gcgactctag agtgagggtc cccacctggg acccttgaga gtatcaggtc
2401 tcccacgtgg gagacaagaa atccctgttt aatatttaaa cagcagtgtt ccccatctgg
2461 gtccttgcac ccctcactct ggcctcagcc gactgcacag cggcccctgc atccccttgg
2521 ctgtgaggcc cctggacaag cagaggtggc cagagctggg aggcatggcc ctggggtccc
2581 acgaatttgc tggggaatct cgtttttctt cttaagactt ttgggacatg gtttgactcc
2641 cgaacatcac cgacgcgtct cctgtttttc tgggtggcct cgggacacct gccctgcccc
2701 cacgagggtc aggactgtga ctctttttag ggccaggcag gtgcctggac atttgccttg
2761 ctggacgggg actggggatg tgggagggag cagacaggag gaatcatgtc aggcctgtgt
2821 gtgaaaggaa gctccactgt caccctccac ctcttcaccc cccactcacc agtgtcccct
2881 ccactgtcac attgtaactg aacttcagga taataaagtg tttgcctcca aaaaaaaaaa
2941 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
3001 aaaaaaaaaa aaaaaaaaag aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
3061 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
3121 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
3181 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
3241 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
3301 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
3361 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
3421 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
3481 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
3541 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta
3601 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
3661 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
3721 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
3781 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
3841 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
3901 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
3961 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
4021 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
4081 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
4141 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
4201 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
4261 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
4321 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
4381 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
4441 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
4501 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
4561 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
4621 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
4681 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
4741 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
4801 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
4861 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
4921 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
4981 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
5041 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
5101 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
5161 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
5221 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
5281 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
5341 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
5401 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
5461 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
5521 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
5581 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
5641 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
5701 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
5761 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
5821 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
5881 gagcgaggaa gc
//
