Basic Vector Information
IKKalpha (S176E,S180E)-pcw107-V5 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 ℃.
IKKalpha (S176E,S180E)-pcw107-V5 vector Sequence
LOCUS IKKalpha_(S176E, 9880 bp DNA circular SYN 13-MAY-2021 DEFINITION when used to produce lentivirus, express physiological levels of insert . ACCESSION . VERSION . KEYWORDS IKKalpha (S176E,S180E)-pcw107-V5. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9880) AUTHORS Martz CA, Ottina KA, Singleton KR, Jasper JS, Wardell SE, Peraza-Penton A, Anderson GR, Winter PS, Wang T, Alley HM, Kwong LN, Cooper ZA, Tetzlaff M, Chen PL, Rathmell JC, Flaherty KT, Wargo JA, McDonnell DP, Sabatini DM, Wood KC TITLE Systematic identification of signaling pathways with potential to confer anticancer drug resistance. JOURNAL Sci Signal. 2014 Dec 23;7(357):ra121. doi: 10.1126/scisignal.aaa1877. PUBMED 25538079 REFERENCE 2 (bases 1 to 9880) TITLE Direct Submission REFERENCE 3 (bases 1 to 9880) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Sci Signal."; date: "2014-12-23"; pages: " 10.1126/scisignal.aaa1877" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..9880 /mol_type="other DNA" /organism="synthetic DNA construct" LTR 1..181 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 228..353 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 846..1079 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1264..1308 /label=gp41 peptide /note="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" promoter 1539..1749 /label=SV40 promoter /note="SV40 early promoter" CDS 1789..2385 /label=PuroR /note="puromycin N-acetyltransferase" misc_feature 2434..2551 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2606..3109 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" protein_bind 3130..3154 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 3172..5406 /gene="CHUK" /label=CHUK /note="Inhibitor of nuclear factor kappa-B kinase subunit alpha from Homo sapiens. Accession#: O15111" protein_bind complement(5411..5431) /label=attB2 /note="core recombination site for the Gateway(R) BP reaction" CDS 5440..5481 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" misc_feature 5506..6094 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 6166..6399 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 6477..6611 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 6638..6773 /label=SV40 ori /note="SV40 origin of replication" promoter complement(6794..6812) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6822..6838) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 6980..7435 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 7461..7565 /label=AmpR promoter CDS 7566..8423 /label=AmpR /note="beta-lactamase" rep_origin 8597..9185 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9339..9356 /label=L4440 /note="L4440 vector, forward primer" protein_bind 9473..9494 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 9509..9539 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9547..9563 /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)." primer_bind 9571..9587 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 9608..9626 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 9654..9880 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter"
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