Basic Vector Information
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | PG13-luc (wt p53 binding sites) | Antibiotic Resistance | Ampicillin |
| Length | 6038 bp | Type | Mammalian Expression, Luciferase |
| 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 Exported 6038 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Luciferase reporter containing 13 copies of the p53-binding
consensus sequence.
ACCESSION .
VERSION .
KEYWORDS PG13-luc (wt p53 binding sites)
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)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 1993
Nov 19. 75(4):817-25."
FEATURES Location/Qualifiers
source 1..6038
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(421..437)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
CDS 491..2143
/codon_start=1
/gene="luc"
/product="firefly luciferase"
/label=luciferase
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
primer_bind complement(557..575)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
intron 2274..2339
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 2469..2489
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 2914..3048
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2951..2970)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3005..3024
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3045..3061)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
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"
primer_bind complement(3137..3153)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3150..3172)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3161..3177
/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(3185..3215)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3230..3251
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/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"
primer_bind complement(3619..3638)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4298..5158)
/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 4921..4940
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5159..5263)
/gene="bla"
/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 complement(5426..5447)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5638..5657
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5871..5893
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 5885..5902
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 5886..5902
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5909..5928
/label=T7
/note="T7 promoter, forward primer"
promoter 5909..5927
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 5952..5968
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 5953..5969
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
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