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PG13-luc (wt p53 binding sites) vector (V012574#)

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

SK primer luciferase small t intron SV40 NLS SV40 poly(A) signal T3 promoter M13 rev lac operator lac promoter CAP binding site L4440 ori AmpR AmpR promoter f1 ori M13/pUC Forward T7 pBluescriptKS

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

Copy Sequence

Download GeneBank File(.gb)

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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