MG15-luc (mut p53 binding sites) vector (V000003)

Basic Vector Information

      • Vector Name:
      • MG15-luc (mut p53 binding sites)
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 5961 bp
      • Type:
      • Mammalian Expression, Luciferase
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • M13pUC-Fwd
      • 3' Primer:
      • LucNRev

MG15-luc (mut p53 binding sites) vector Vector Map

MG15-luc (mut p53 binding sites)5961 bp60012001800240030003600420048005400SK primerluciferasesmall t intronSV40 NLSSV40 poly(A) signalT3 promoterM13 revlac operatorlac promoterCAP binding siteL4440oriAmpRAmpR promoterf1 oriIn lacZ geneT7pBluescriptKS

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 ℃.

MG15-luc (mut p53 binding sites) vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                5961 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    MG15-luc (mut p53 binding sites)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5961)
  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 5961)
  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..5961
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(410..426)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             480..2132
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(546..564)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          2263..2328
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2458..2478
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2903..3037
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2940..2959)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2994..3013
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3034..3050)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3087..3105)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3126..3142)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3126..3142)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3139..3161)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3150..3166
                     /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(3174..3204)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3219..3240
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3357..3374)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3528..4116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3608..3627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4287..5147)
                     /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     4910..4929
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5148..5252)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5278..5733)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5415..5436)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5627..5646
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5860..5882
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5874..5891
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5875..5891
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5898..5917
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5898..5916
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     5941..5957
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5942..5958
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"

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