IgK-IFN-luc vector (V010735)

Basic Vector Information

      • Vector Name:
      • IgK-IFN-luc
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 5863 bp
      • Type:
      • Mammalian Expression, Luciferase
      • Copy Number:
      • High Copy
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • na
      • 3' Primer:
      • LucNrev

IgK-IFN-luc vector Vector Map

IgK-IFN-luc5863 bp60012001800240030003600420048005400T3 promoterM13 revlac operatorlac promoterCAP binding siteL4440oriAmpRAmpR promoterf1 oriIn lacZ geneT7SV40 poly(A) signalluciferasesmall t intronSV40 NLSSV40 poly(A) signal

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 ℃.

IgK-IFN-luc vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                5863 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Reporter construct for NF‐kappa-B activation. Contains two copies of
            immunoglobulin kappa light chain motif (5′‐GGGGACTTTCC‐3′) upstream 
            of the a minimal promoter driving luciferase expression.
ACCESSION   .
VERSION     .
KEYWORDS    IgK-IFN-luc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5863)
  AUTHORS   Pomerantz JL, Denny EM, Baltimore D
  TITLE     CARD11 mediates factor-specific activation of NF-kappaB by the T 
            cell receptor complex.
  JOURNAL   EMBO J. 2002 Oct 1. 21(19):5184-94.
  PUBMED    12356734
REFERENCE   2  (bases 1 to 5863)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "EMBO J. 
            2002 Oct 1. 21(19):5184-94."
FEATURES             Location/Qualifiers
     source          1..5863
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(33..51)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(72..88)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(72..88)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(85..107)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    96..112
                     /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(120..150)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    165..186
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(303..320)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(474..1062)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(554..573)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1233..2093)
                     /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     1856..1875
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2094..2198)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2224..2679)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2361..2382)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2573..2592
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2806..2828
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2820..2837
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2821..2837
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2844..2863
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2844..2862
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2872..3006
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             3210..4862
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(3276..3294)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          5086..5151
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5281..5301
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5726..5860
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"

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