| 载体基本信息 |
| 载体名称 |
GFP-TAF1 |
载体抗性 |
Ampicillin |
| 载体长度 |
11569 bp |
载体类型 |
Mammalian Expression |
| 筛选标记 |
Blasticidin |
拷贝数 |
Low Copy |
| 启动子 |
CMV |
克隆方法 |
Gateway Cloning |
| 5引物 |
EGFP_C_primer |
3引物 |
TK polyA reverse primer |
GFP-TAF1 载体载体图谱
GFP-TAF1 载体载体序列
LOCUS Exported 11569 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION GFP-tagged BRD protein, which can be used to be expressed in
mammalian cells..
ACCESSION .
VERSION .
KEYWORDS GFP-TAF1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11569)
AUTHORS Gong F, Chiu LY, Cox B, Aymard F, Clouaire T, Leung JW, Cammarata M,
Perez M, Agarwal P, Brodbelt JS, Legube G, Miller KM
TITLE Screen identifies bromodomain protein ZMYND8 in chromatin
recognition of transcription-associated DNA damage that promotes
homologous recombination.
JOURNAL Genes Dev. 2015 Jan 15;29(2):197-211. doi: 10.1101/gad.252189.114.
PUBMED 25593309
REFERENCE 2 (bases 1 to 11569)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1101/gad.252189"; journalName: "Genes Dev"; date: "2015-01-15-
15"; volume: "29"; issue: "2"; pages: "197-211"
FEATURES Location/Qualifiers
source 1..11569
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(11..28)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(182..770)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(262..281)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(941..1801)
/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 1564..1583
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1802..1906)
/gene="bla"
/label=AmpR promoter
primer_bind complement(1981..2000)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 2172..2551
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2552..2755
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2706..2726
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 2752..2776
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 2880..3596
/codon_start=1
/product="Emerald GFP"
/label=EmGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(2925..2946)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3186..3205)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3533..3554
/label=EGFP-C
/note="EGFP, forward primer"
protein_bind 3612..3636
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(9331..9355)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 9363..9404
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
primer_bind complement(9444..9463)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 9488..9536
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 9738..10166
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(9825..9844)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 10035..10056
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(10175..10195)
/label=pBABE 3
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 10180..10509
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 10360..10495
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 10422..10441
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 10557..10604
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 10623..11021
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
polyA_signal 11179..11300
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(11216..11235)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 11270..11289
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(11349..11365)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(11349..11365)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(11362..11384)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 11373..11389
/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(11397..11427)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 11442..11463
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."