Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000986 | IKKalpha (S176E,S180E)-pcw107-V5 | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- IKKalpha (S176E,S180E)-pcw107-V5
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9880 bp
- Type:
- Lentiviral
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Promoter:
- hPGK
- Cloning Method:
- Gateway Cloning
- 5' Primer:
- hPGK-F
- 3' Primer:
- WPRE-R
IKKalpha (S176E,S180E)-pcw107-V5 vector Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
IKKalpha (S176E,S180E)-pcw107-V5 vector Sequence
LOCUS V000986 9880 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000986
VERSION V000986
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"
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="Inhibitor of nuclear factor kappa-B kinase subunit
alpha"
/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"