| 质粒名称: | pEBB-TB |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 36099 |
| 存储实验室: | Ezra Burstein |
|
载体骨架基本信息 |
|
| 载体名称: | pEBB |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5450 bp |
| 载体修饰: | BT-tag（P.freudenreichia-transcarboxilase的生物素化位点，通过TEV位点与MCS分离） |
| 载体类型: | 哺乳细胞表达 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 启动子 | EF1a |
| 标签/融合蛋白: | BT-tag（插入C端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 测序引物 | CTCAAGCCTCAGACAGTGG |
| 3' 测序引物 | GAGGGGTGCAAGGACCCAG |
|
文献引用方法 |
|
| 材料和方法部分: | pEBB-TB was a gift from Ezra Burstein (Addgene plasmid # 36099 ; http://n2t.net/addgene:36099 ; RRID:Addgene_36099) |
| 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
pEBBTB载体基本信息
pEBBTB载体图谱质粒图谱和多克隆位点信息
pEBBTB载体简介
pEBBTB载体序列
LOCUS pEBB-TB 5678 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pEBB-TB
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5678)
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 5678)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5678
/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"
CDS 1841..1861
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
primer_bind complement(2812..2829)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(2812..2828)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2821..2843)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3041..3496
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3128..3147)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3338..3359
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3513..3532)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3632..3654
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(3692..3710)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3778..3882
/gene="bla"
/label=AmpR promoter
CDS 3883..4743
/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(4101..4120)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4914..5502
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5403..5422
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5656..5673
/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 gaattctcta gagatccctc gacctcgaga tccattgtgc tggatccact agtggtacca
1801 tcgatgcggc cgcgactacg atatacccac aaccgccagc gagaatctgt actttcaatc
1861 agagttaaaa accgccgcac tagctcaaca tgacgaggct gctggaaagg ccggtgaagg
1921 tgaaatccct gcccctcttg ctggtaccgt ttctaagata ctggtaaaag aaggtgacac
1981 tgttaaagct ggtcaaacag ttctggtgct ggaggctatg aaaatggaga cagaaattaa
2041 cgctcctact gacggaaaag ttgaaaaggt gttagttaag gaaagagatg ctgttcaagg
2101 tggtcaaggt ctaatcaaga tcggcgttta agggccgcga ctctagagtg agggtcccca
2161 cctgggaccc ttgagagtat caggtctccc acgtgggaga caagaaatcc ctgtttaata
2221 tttaaacagc agtgttcccc atctgggtcc ttgcacccct cactctggcc tcagccgact
2281 gcacagcggc ccctgcatcc ccttggctgt gaggcccctg gacaagcaga ggtggccaga
2341 gctgggaggc atggccctgg ggtcccacga atttgctggg gaatctcgtt tttcttctta
2401 agacttttgg gacatggttt gactcccgaa catcaccgac gcgtctcctg tttttctggg
2461 tggcctcggg acacctgccc tgcccccacg agggtcagga ctgtgactct ttttagggcc
2521 aggcaggtgc ctggacattt gccttgctgg acggggactg gggatgtggg agggagcaga
2581 caggaggaat catgtcaggc ctgtgtgtga aaggaagctc cactgtcacc ctccacctct
2641 tcacccccca ctcaccagtg tcccctccac tgtcacattg taactgaact tcaggataat
2701 aaagtgtttg cctccaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
2761 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaagaatt cactggccgt
2821 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
2881 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
2941 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
3001 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
3061 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
3121 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
3181 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
3241 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
3301 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
3361 acaacactca accctatctc gggctattct tttgatttat aagggatttt gccgatttcg
3421 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
3481 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
3541 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
3601 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
3661 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
3721 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
3781 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
3841 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
3901 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
3961 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
4021 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
4081 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
4141 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
4201 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
4261 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
4321 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
4381 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
4441 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
4501 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
4561 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
4621 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
4681 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
4741 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
4801 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
4861 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
4921 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
4981 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
5041 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
5101 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
5161 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
5221 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
5281 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
5341 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
5401 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
5461 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
5521 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
5581 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
5641 cgaacgaccg agcgcagcga gtcagtgagc gaggaagc
//
