| 质粒名称: | pEBB HA RelA |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 74892 |
| 存储实验室: | Ezra Burstein, Colin Duckett |
|
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: | 5350 bp |
| 载体类型: | 哺乳细胞表达 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | RelA |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1656 bp |
| GenBank编号 | NM_021975 |
| 启动子 | EF-1alpha promoter |
| 标签/融合蛋白: | HA（主干上的N终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI (未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CTC AAG CCT CAG ACA GTG G |
| 3' 测序引物 | GAG GGG TGC AAG GAC CCA G |
|
文献引用方法 |
|
| 材料和方法部分: | pEBB HA RelA was a gift from Ezra Burstein & Colin Duckett (Addgene plasmid # 74892 ; http://n2t.net/addgene:74892 ; RRID:Addgene_74892) |
| 参考文献部分: | COMMD proteins, a novel family of structural and functional homologs of MURR1. Burstein E, Hoberg JE, Wilkinson AS, Rumble JM, Csomos RA, Komarck CM, Maine GN, Wilkinson JC, Mayo MW, Duckett CS. J Biol Chem. 2005 Jun 10;280(23):22222-32. Epub 2005 Mar 30. 10.1074/jbc.M501928200 PubMed 15799966 |
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 HA RelA 7046 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Express RelA in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pEBB HA RelA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7046)
AUTHORS Burstein E, Hoberg JE, Wilkinson AS, Rumble JM, Csomos RA, Komarck
CM, Maine GN, Wilkinson JC, Mayo MW, Duckett CS
TITLE COMMD proteins, a novel family of structural and functional homologs
of MURR1.
JOURNAL J Biol Chem. 2005 Jun 10;280(23):22222-32. Epub 2005 Mar 30.
PUBMED 15799966
REFERENCE 2 (bases 1 to 7046)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7046
/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 1804..1830
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 3124..3495
/codon_start=1
/product="transcriptional activation domain of human RelA,
also known as p65 (O'Shea and Perkins, 2008)"
/label=RelA (p65) AD
/translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
EDFSSIADMDFSALLSQISS"
primer_bind complement(4180..4197)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4180..4196)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4189..4211)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 4409..4864
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4496..4515)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4706..4727
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4881..4900)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5000..5022
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5060..5078)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5146..5250
/gene="bla"
/label=AmpR promoter
CDS 5251..6111
/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(5469..5488)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6282..6870
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6771..6790
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7024..7041
/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 tggatctcac catggcttct
1801 agctatcctt atgacgtgcc tgactatgcc agcctgggat ccatggacga actgttcccc
1861 ctcatcttcc cggcagagcc agcccaggcc tctggcccct atgtggagat cattgagcag
1921 cccaagcagc ggggcatgcg cttccgctac aagtgcgagg ggcgctccgc gggcagcatc
1981 ccaggcgaga ggagcacaga taccaccaag acccacccca ccatcaagat caatggctac
2041 acaggaccag ggacagtgcg catctccctg gtcaccaagg accctcctca ccggcctcac
2101 ccccacgagc ttgtaggaaa ggactgccgg gatggcttct atgaggctga gctctgcccg
2161 gaccgctgca tccacagttt ccagaacctg ggaatccagt gtgtgaagaa gcgggacctg
2221 gagcaggcta tcagtcagcg catccagacc aacaacaacc ccttccaagt tcctatagaa
2281 gagcagcgtg gggactacga cctgaatgct gtgcggctct gcttccaggt gacagtgcgg
2341 gacccatcag gcaggcccct ccgcctgccg cctgtccttt ctcatcccat ctttgacaat
2401 cgtgccccca acactgccga gctcaagatc tgccgagtga accgaaactc tggcagctgc
2461 ctcggtgggg atgagatctt cctactgtgt gacaaggtgc agaaagagga cattgaggtg
2521 tatttcacgg gaccaggctg ggaggcccga ggctcctttt cgcaagctga tgtgcaccga
2581 caagtggcca ttgtgttccg gacccctccc tacgcagacc ccagcctgca ggctcctgtg
2641 cgtgtctcca tgcagctgcg gcggccttcc gaccgggagc tcagtgagcc catggaattc
2701 cagtacctgc cagatacaga cgatcgtcac cggattgagg agaaacgtaa aaggacatat
2761 gagaccttca agagcatcat gaagaagagt cctttcagcg gacccaccga cccccggcct
2821 ccacctcgac gcattgctgt gccttcccgc agctcagctt ctgtccccaa gccagcaccc
2881 cagccctatc cctttacgtc atccctgagc accatcaact atgatgagtt tcccaccatg
2941 gtgtttcctt ctgggcagat cagccaggcc tcggccttgg ccccggcccc tccccaagtc
3001 ctgccccagg ctccagcccc tgcccctgct ccagccatgg tatcagctct ggcccaggcc
3061 ccagcccctg tcccagtcct agccccaggc cctcctcagg ctgtggcccc acctgccccc
3121 aagcccaccc aggctgggga aggaacgctg tcagaggccc tgctgcagct gcagtttgat
3181 gatgaagacc tgggggcctt gcttggcaac agcacagacc cagctgtgtt cacagacctg
3241 gcatccgtcg acaactccga gtttcagcag ctgctgaacc agggcatacc tgtggccccc
3301 cacacaactg agcccatgct gatggagtac cctgaggcta taactcgcct agtgacaggg
3361 gcccagaggc cccccgaccc agctcctgct ccactggggg ccccggggct ccccaatggc
3421 ctcctttcag gagatgaaga cttctcctcc attgcggaca tggacttctc agccctgctg
3481 agtcagatca gctcctaagc ggccgcgact ctagagtgag ggtccccacc tgggaccctt
3541 gagagtatca ggtctcccac gtgggagaca agaaatccct gtttaatatt taaacagcag
3601 tgttccccat ctgggtcctt gcacccctca ctctggcctc agccgactgc acagcggccc
3661 ctgcatcccc ttggctgtga ggcccctgga caagcagagg tggccagagc tgggaggcat
3721 ggccctgggg tcccacgaat ttgctgggga atctcgtttt tcttcttaag acttttggga
3781 catggtttga ctcccgaaca tcaccgacgc gtctcctgtt tttctgggtg gcctcgggac
3841 acctgccctg cccccacgag ggtcaggact gtgactcttt ttagggccag gcaggtgcct
3901 ggacatttgc cttgctggac ggggactggg gatgtgggag ggagcagaca ggaggaatca
3961 tgtcaggcct gtgtgtgaaa ggaagctcca ctgtcaccct ccacctcttc accccccact
4021 caccagtgtc ccctccactg tcacattgta actgaacttc aggataataa agtgtttgcc
4081 tccaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
4141 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaagaattca ctggccgtcg ttttacaacg
4201 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
4261 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
4321 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
4381 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
4441 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
4501 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
4561 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
4621 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
4681 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
4741 cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
4801 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
4861 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
4921 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
4981 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
5041 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
5101 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
5161 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
5221 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
5281 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
5341 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
5401 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
5461 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
5521 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
5581 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
5641 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
5701 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
5761 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
5821 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
5881 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
5941 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
6001 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
6061 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
6121 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
6181 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
6241 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
6301 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
6361 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
6421 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
6481 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
6541 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
6601 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
6661 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
6721 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
6781 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
6841 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
6901 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
6961 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
7021 cgcagcgagt cagtgagcga ggaagc
//
