PPRE X3-TK-luc vector (V012167)

Basic Vector Information

      • Vector Name:
      • PPRE X3-TK-luc
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 6763 bp
      • Type:
      • Mammalian Expression, Luciferase
      • Copy Number:
      • High Copy
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • M13pUC-fwd
      • 3' Primer:
      • LucNrev

PPRE X3-TK-luc vector Vector Map

PPRE X3-TK-luc6763 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600M13 ForwardM13 oripRS-markerpGEX 3'pBRforEcoAmpR promoterAmpRoriL4440CAP binding sitelac promoterlac operatorM13 revSV40 poly(A) signalSV40 NLSsmall t intronluciferaseMini-TK promoter

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 ℃.

PPRE X3-TK-luc vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6763 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  PPAR reporter.  3xDR1 sites upstream of a luciferase reporter..
ACCESSION   .
VERSION     .
KEYWORDS    PPRE X3-TK-luc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6763)
  AUTHORS   Kim JB, Wright HM, Wright M, Spiegelman BM
  TITLE     ADD1/SREBP1 activates PPARgamma through the production of endogenous
            ligand.
  JOURNAL   Proc Natl Acad Sci U S A 1998 Apr 14;95(8):4333-7.
  PUBMED    9539737
REFERENCE   2  (bases 1 to 6763)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A"; date: "1998-04-14"; volume: "95(8)"; pages: 
            "4333-7"
FEATURES             Location/Qualifiers
     source          1..6763
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(274..291)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(274..290)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(283..305)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      731..1244
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(818..837)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1028..1049
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1726..1745)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1845..1867
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1905..1923)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1991..2095
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2096..2956
                     /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     complement(2314..2333)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3127..3715
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3616..3635
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3869..3886
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4003..4024
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4039..4069
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4077..4093
                     /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     4082..4104
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4101..4117
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4101..4117
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4135..4269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4159..4178)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4213..4232
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4694..4714)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4844..4909
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(5040..6692)
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     6608..6626
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     promoter        complement(join(6752..6763,1..51))
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"

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