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GFP-TAF1 vector (V011373#)

Basic Vector Information

Basic Vector Information
Vector Name GFP-TAF1 Antibiotic Resistance Ampicillin
Length 11569 bp Type Mammalian Expression
Selection Marker Blasticidin Copy Number Low Copy
Promoter CMV Cloning Method Gateway Cloning
5' Primer EGFP_C_primer 3' Primer TK polyA reverse primer

GFP-TAF1 vector Vector Map

GFP-TAF111569 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000105001100011500L4440oriAmpRAmpR promoterpRS-markerCMV enhancerCMV promoterEmGFPattB1attB2V5 tagTK-pA-RHSV TK poly(A) signalf1 oripBABE 3'EM7 promoterBSDSV40 poly(A) signalM13 revlac operatorlac promoterCAP binding site

Plasmid Resuspension protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

GFP-TAF1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

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."

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