Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V011373 | GFP-TAF1 | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- GFP-TAF1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11569 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- 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 Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
GFP-TAF1 vector Sequence
LOCUS 40924_1059 11569 bp DNA circular SYN 13-MAY-2021
DEFINITION GFP-tagged BRD protein, which can be used to be expressed in
mammalian cells..
ACCESSION .
VERSION .
KEYWORDS .
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)
TITLE Direct Submission
REFERENCE 3 (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"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11569
/mol_type="other DNA"
/organism="synthetic DNA construct"
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"
CDS complement(944..1801)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(1802..1906)
/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 2752..2776
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 2880..3596
/codon_start=1
/label=EmGFP
/note="Emerald GFP"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
protein_bind 3612..3636
/label=attB1
/note="recombination site for the Gateway(R) BP reaction"
CDS complement(5197..5211)
/codon_start=1
/label=SNAC tag
/note="tag for protein cleavage using Ni2+ ion (Dang et
al., 2019)"
/translation="GSHHW"
protein_bind complement(9331..9355)
/label=attB2
/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"
promoter 10180..10509
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 10557..10604
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 10623..11018
/codon_start=1
/label=BSD
/note="blasticidin S deaminase"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
polyA_signal 11179..11312
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
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 complement(11442..11463)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."