WWP-Luc (p21/WAF1 promoter) vector (V006865)

Basic Vector Information

      • Vector Name:
      • WWP-Luc (p21/WAF1 promoter)
      • Antibiotic Resistance:
      • Chloramphenicol
      • Length:
      • 8366 bp
      • Type:
      • Mammalian Expression, Luciferase
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • T7

WWP-Luc (p21/WAF1 promoter) vector Vector Map

WWP-Luc (p21/WAF1 promoter)8366 bp400800120016002000240028003200360040004400480052005600600064006800720076008000CAP binding sitelac promoterlac operatorM13 revT3SV40 poly(A) signalSV40 NLSsmall t intronluciferaseSK primerKS primerSK primerT7M13 Forwardf1 oriAmpR promotercat promoterCmRoriL4440

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 ℃.

WWP-Luc (p21/WAF1 promoter) vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                8366 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    WWP-Luc (p21/WAF1 promoter)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8366)
  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 8366)
  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..8366
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     240..260
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    291..425
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(315..334)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     369..388
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(850..870)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          1000..1065
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(1196..2848)
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     2764..2782
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     2902..2918
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2917..2933
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2918..2934
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5306..5322)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5354..5373)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5355..5373)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5383..5400)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5383..5399)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5392..5414)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5541..5996
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5628..5647)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5838..5859
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6022..6126
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6446..6548
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             6549..7208
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6615..6634)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      7597..8185
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8086..8105
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8339..8356
                     /label=L4440
                     /note="L4440 vector, forward primer"

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