Basic Vector Information
- Vector Name:
- IgK-IFN-luc
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5863 bp
- Type:
- Mammalian Expression, Luciferase
- Replication origin:
- ori
- Copy Number:
- High Copy
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- na
- 3' Primer:
- LucNrev
IgK-IFN-luc vector Vector Map
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
LOCUS 40924_1404 5863 bp 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 . 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) TITLE Direct Submission REFERENCE 3 (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." COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..5863 /mol_type="other DNA" /organism="synthetic DNA construct" 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" protein_bind complement(96..112) /label=lac operator /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 complement(165..186) /label=CAP binding site /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" CDS complement(1236..2093) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(2094..2198) /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 2821..2837 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" 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..4859 /codon_start=1 /label=luciferase /note="firefly luciferase" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL" intron 5086..5151 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 5281..5301 /codon_start=1 /label=SV40 NLS /note="nuclear localization signal of SV40 (simian virus 40) large T antigen" /translation="PKKKRKV" polyA_signal 5726..5860 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal"
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