| 质粒名称: | pFlag-CMV2-Brd4 (1-1362) |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 22304 |
| 存储实验室: | Eric Verdin |
|
载体骨架基本信息 |
|
| 载体名称: | pFLAG-CMV2 |
| 载体出品公司: | Sigma |
| 空载体大小: | 4700 bp |
| 完整质粒大小: | 8729bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | BRD4 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 4100 bp |
| 标签/融合蛋白: | Flag (N terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | HindIII(未破坏) |
| 3’克隆位点: | BamHI (未破坏) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | hGH-PA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pFlag-CMV2-Brd4 (1-1362) was a gift from Eric Verdin (Addgene plasmid # 22304) |
| 参考文献部分: | Conserved P-TEFb-interacting domain of BRD4 inhibits HIV transcription. Bisgrove DA, Mahmoudi T, Henklein P, Verdin E. Proc Natl Acad Sci U S A. 2007 Aug 21. 104(34):13690-5. 10.1073/pnas.0705053104 PubMed 17690245 |
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
质粒图谱.png)
载体简介
载体序列
LOCUS pFlag-CMV2-Brd4 (1-1362) 8729 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFlag-CMV2-Brd4 (1-1362)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8729)
AUTHORS Bisgrove DA, Mahmoudi T, Henklein P, Verdin E
TITLE Conserved P-TEFb-interacting domain of BRD4 inhibits HIV
transcription.
JOURNAL Proc Natl Acad Sci U S A. 2007 Aug 21. 104(34):13690-5.
PUBMED 17690245
REFERENCE 2 (bases 1 to 8729)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8729
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(63..80)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(234..822)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(314..333)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(993..1853)
/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 1616..1635
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1854..1958)
/gene="bla"
/label=AmpR promoter
rep_origin 1985..2440
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2072..2091)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2282..2303
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2566..2588
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2580..2597
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2581..2597
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
enhancer 2758..3137
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3138..3341
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 3292..3312
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 3375..3398
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 5613..5630
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
polyA_signal 7510..8132
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(7709..7728)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
primer_bind complement(8156..8176)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 8161..8490
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 8341..8476
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 8403..8422
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(8528..8547)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(8529..8547)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(8561..8577)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8561..8577)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8574..8596)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8585..8601
/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)."
promoter complement(8609..8639)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8654..8675
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
ORIGIN
1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
1981 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
2041 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
2101 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
2161 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
2221 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
2281 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
2341 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
2401 tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca ttcaggctgc
2461 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
2521 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
2581 gtaaaacgac ggccagtgcc aagctgatct atacattgaa tcaatattgg caattagcca
2641 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
2701 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac
2761 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
2821 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
2881 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
2941 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
3001 tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
3061 attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag
3121 tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt
3181 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
3241 accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg
3301 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
3361 attgatctac catggactac aaagacgatg acgacaagct tatgtctgcg gagagcggcc
3421 ctgggacgag attgagaaat ctgccagtaa tgggggatgg actagaaact tcccaaatgt
3481 ctacaacaca ggcccaggcc caaccccagc cagccaacgc agccagcacc aaccccccgc
3541 ccccagagac ctccaaccct aacaagccca agaggcagac caaccaactg caatacctgc
3601 tcagagtggt gctcaagaca ctatggaaac accagtttgc atggcctttc cagcagcctg
3661 tggatgccgt caagctgaac ctccctgatt actataagat cattaaaacg cctatggata
3721 tgggaacaat aaagaagcgc ttggaaaaca actattactg gaatgctcag gaatgtatcc
3781 aggacttcaa cactatgttt acaaattgtt acatctacaa caagcctgga gatgacatag
3841 tcttaatggc agaagctctg gaaaagctct tcttgcaaaa aataaatgag ctacccacag
3901 aagaaaccga gatcatgata gtccaggcaa aaggaagagg acgtgggagg aaagaaacag
3961 ggacagcaaa acctggcgtt tccacggtac caaacacaac tcaagcatcg actcctccgc
4021 agacccagac ccctcagccg aatcctcctc ctgtgcaggc cacgcctcac cccttccctg
4081 ccgtcacccc ggacctcatc gtccagaccc ctgtcatgac agtggtgcct ccccagccac
4141 tgcagacgcc cccgccagtg cccccccagc cacaaccccc acccgctcca gctccccagc
4201 ccgtacagag ccacccaccc atcatcgcgg ccaccccaca gcctgtgaag acaaagaagg
4261 gagtgaagag gaaagcagac accaccaccc ccaccaccat tgaccccatt cacgagccac
4321 cctcgctgcc cccggagccc aagaccacca agctgggcca gcggcgggag agcagccggc
4381 ccgtgaaacc tccaaagaag gacgtgcccg actctcagca gcacccagca ccagagaaga
4441 gcagcaaggt ctcggagcag ctcaagtgct gcagcggcat cctcaaggag atgtttgcca
4501 agaagcacgc cgcctacgcc tggcccttct acaagcctgt ggacgtggag gcactgggcc
4561 tacacgacta ctgtgacatc atcaagcacc ccatggacat gagcacaatc aagtctaaac
4621 tggaggcccg tgagtaccgt gatgctcagg agtttggtgc tgacgtccga ttgatgttct
4681 ccaactgcta taagtacaac cctcctgacc atgaggtggt ggccatggcc cgcaagctcc
4741 aggatgtgtt cgaaatgcgc tttgccaaga tgccggacga gcctgaggag ccagtggtgg
4801 ccgtgtcctc cccggcagtg ccccctccca ccaaggttgt ggccccgccc tcatccagcg
4861 acagcagcag cgatagctcc tcggacagtg acagttcgac tgatgactct gaggaggagc
4921 gagcccagcg gctggctgag ctccaggagc agctcaaagc cgtgcacgag cagcttgcag
4981 ccctctctca gccccagcag aacaaaccaa agaaaaagga gaaagacaag aaggaaaaga
5041 aaaaagaaaa gcacaaaagg aaagaggaag tggaagagaa taaaaaaagc aaagccaagg
5101 aacctcctcc taaaaagacg aagaaaaata atagcagcaa cagcaatgtg agcaagaagg
5161 agccagcgcc catgaagagc aagccccctc ccacgtatga gtcggaggaa gaggacaagt
5221 gcaagcctat gtcctatgag gagaagcggc agctcagctt ggacatcaac aagctccccg
5281 gcgagaagct gggccgcgtg gtgcacatca tccagtcacg ggagccctcc ctgaagaatt
5341 ccaaccccga cgagattgaa atcgactttg agaccctgaa gccgtccaca ctgcgtgagc
5401 tggagcgcta tgtcacctcc tgtttgcgga agaaaaggaa acctcaagct gagaaagttg
5461 atgtgattgc cggctcctcc aagatgaagg gcttctcgtc ctcagagtcg gagagctcca
5521 gtgagtccag ctcctctgac agcgaagact ccgaaacaga gatggctccg aagtcaaaaa
5581 agaaggggca ccccgggagg gagcagaaga agcaccatca tcaccaccat cagcagatgc
5641 agcaggcccc ggctcctgtg ccccagcagc cgcccccgcc tccccagcag cccccaccgc
5701 ctccacctcc gcagcagcaa cagcagccgc cacccccgcc tcccccaccc tccatgccgc
5761 agcaggcagc cccggcgatg aagtcctcgc ccccaccctt cattgccacc caggtgcccg
5821 tcctggagcc ccagctccca ggcagcgtct ttgaccccat cggccacttc acccagccca
5881 tcctgcacct gccgcagcct gagctgcccc ctcacctgcc ccagccgcct gagcacagca
5941 ctccacccca tctcaaccag cacgcagtgg tctctcctcc agctttgcac aacgcactac
6001 cccagcagcc atcacggccc agcaaccgag ccgctgccct gcctcccaag cccgcccggc
6061 ccccagccgt gtcaccagcc ttgacccaaa cacccctgct cccacagccc cccatggccc
6121 aaccccccca agtgctgctg gaggatgaag agccacctgc cccacccctc acctccatgc
6181 agatgcagct gtacctgcag cagctgcaga aggtgcagcc ccctacgccg ctactccctt
6241 ccgtgaaggt gcagtcccag cccccacccc ccctgccgcc cccaccccac ccctctgtgc
6301 agcagcagct gcagcagcag ccgccaccac ccccaccacc ccagccccag cctccacccc
6361 agcagcagca tcagccccct ccacggcccg tgcacttgca gcccatgcag ttttccaccc
6421 acatccaaca gcccccgcca ccccagggcc agcagccccc ccatccgccc ccaggccagc
6481 agccaccccc gccgcagcct gccaagcctc agcaagtcat ccagcaccac cattcacccc
6541 ggcaccacaa gtcggacccc tactcaaccg gtcacctccg cgaagccccc tccccgctta
6601 tgatacattc cccccagatg tcacagttcc agagcctgac ccaccagtct ccaccccagc
6661 aaaacgtcca gcctaagaaa caggagctgc gtgctgcctc cgtggtccag ccccagcccc
6721 ttgtggtggt gaaggaggag aagatccact cacccatcat ccgcagcgag cccttcagcc
6781 cctcgctgcg gccggagccc cccaagcacc cggagagcat caaggccccc gtccacctgc
6841 cccagcggcc ggaaatgaag cctgtggatg tcgggaggcc tgtgatccgg cccccagagc
6901 agaacgcacc gccaccaggg gcccctgaca aggacaaaca gaaacaggag ccgaagactc
6961 cagttgcgcc caaaaaggac ctgaaaatca agaacatggg ctcctgggcc agcctagtgc
7021 agaagcatcc gaccaccccc tcctccacag ccaagtcatc cagcgacagc ttcgagcagt
7081 tccgccgcgc cgctcgggag aaagaggagc gtgagaaggc cctgaaggct caggccgagc
7141 acgctgagaa ggagaaggag cggctgcggc aggagcgcat gaggagccga gaggacgagg
7201 atgcgctgga gcaggcccgg cgggcccatg aggaggcacg tcggcgccag gagcagcagc
7261 agcagcagcg ccaggagcaa cagcagcagc agcaacagca agcagctgcg gtggctgccg
7321 ccgccacccc acaggcccag agctcccagc cccagtccat gctggaccag cagagggagt
7381 tggcccggaa gcgggagcag gagcgaagac gccgggaagc catggcagct accattgaca
7441 tgaatttcca gagtgatcta ttgtcaatat ttgaagaaaa tcttttctga gcgcacctag
7501 gtggatcccg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
7561 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
7621 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
7681 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
7741 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
7801 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
7861 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
7921 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
7981 gtccttctga ttttaaaata actataccag caggaggacg tccagacaca gcataggcta
8041 cctggccatg cccaaccggt gggacatttg agttgcttgc ttggcactgt cctctcatgc
8101 gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc ggccagcttg gctgtggaat
8161 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
8221 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
8281 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
8341 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
8401 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
8461 ggcttttttg gaggcctagg cttttgcaaa aagctcctcg aggaactgaa aaaccagaaa
8521 gttaattccc tatagtgagt cgtattaaat tcgtaatcat gtcatagctg tttcctgtgt
8581 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
8641 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
8701 tccagtcggg aaacctgtcg tgccagctg
//
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