Basic Vector Information
- Vector Name:
- PG13-luc (wt p53 binding sites)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6038 bp
- Type:
- Mammalian Expression, Luciferase
- Replication origin:
- ori
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- T7
- 3' Primer:
- LucNrev
PG13-luc (wt p53 binding sites) vector Vector Map
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 ℃.
PG13-luc (wt p53 binding sites) vector Sequence
LOCUS 40924_20830 6038 bp DNA circular SYN 13-MAY-2021
DEFINITION Luciferase reporter containing 13 copies of the p53-binding
consensus sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6038)
AUTHORS el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM,
Lin D, Mercer WE, Kinzler KW, Vogelstein B
TITLE WAF1, a potential mediator of p53 tumor suppression.
JOURNAL Cell. 1993 Nov 19. 75(4):817-25.
PUBMED 8242752
REFERENCE 2 (bases 1 to 6038)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6038)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 1993
Nov 19. 75(4):817-25."
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6038
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(421..437)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
CDS 491..2140
/codon_start=1
/label=luciferase
/note="firefly luciferase"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
intron 2274..2339
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 2469..2489
/codon_start=1
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/translation="PKKKRKV"
polyA_signal 2914..3048
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter complement(3098..3116)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3137..3153)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3161..3177)
/label=lac operator
/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(3185..3215)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3230..3251)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3368..3385)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3539..4127)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4301..5158)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5159..5263)
/label=AmpR promoter
rep_origin complement(5289..5744)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 5886..5902
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 5909..5927
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 5953..5969
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
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