Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V001643 | pLenti6/UbC/mSlc7a1 | 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:
- pLenti6/UbC/mSlc7a1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9570 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Selection Marker:
- Blasticidin
- Copy Number:
- High Copy
- Promoter:
- UbC
- Cloning Method:
- Gateway Cloning
- 5' Primer:
- hUBCpro-F (5'-TGAAGCTCCGGTTTTGAACT-3')
- 3' Primer:
- pBABE3
pLenti6/UbC/mSlc7a1 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.
pLenti6/UbC/mSlc7a1 vector Sequence
LOCUS V001643 9570 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V001643
VERSION V001643
KEYWORDS pLenti6/UbC/mSlc7a1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 9570)
AUTHORS Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K,
Yamanaka S
TITLE Induction of pluripotent stem cells from adult human fibroblasts by
defined factors.
JOURNAL Cell. 2007 Nov 30. 131(5):861-72.
PUBMED 18035408
REFERENCE 2 (bases 1 to 9570)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 9570)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2007
Nov 30. 131(5):861-72."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9570
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 3..229
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 230..410
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 457..582
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1075..1308
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1493..1537
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
CDS 1686..1727
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
promoter 1803..3014
/label="UbC promoter"
/note="human ubiquitin C promoter"
protein_bind 3074..3098
/label="attB1"
/note="recombination site for the Gateway(R) BP reaction"
CDS 3119..4984
/gene="Slc7a1"
/label="High affinity cationic amino acid transporter 1"
/note="High affinity cationic amino acid transporter 1 from
Mus musculus. Accession#: Q09143"
protein_bind complement(5003..5027)
/label="attB2"
/note="recombination site for the Gateway(R) BP reaction"
CDS 5080..5121
/label="V5 tag"
/note="epitope tag from simian virus 5"
promoter 5163..5492
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
promoter 5540..5587
/label="EM7 promoter"
/note="synthetic bacterial promoter"
CDS 5606..6001
/label="BSD"
/note="blasticidin S deaminase"
LTR 6091..6324
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 6396..6530
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 6557..6692
/label="SV40 ori"
/note="SV40 origin of replication"
promoter complement(6713..6731)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6741..6757)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 6899..7354
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 7380..7484
/label="AmpR promoter"
CDS 7485..8342
/label="AmpR"
/note="beta-lactamase"
rep_origin 8516..9104
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9258..9275
/label="L4440"
/note="L4440 vector, forward primer"
protein_bind 9392..9413
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 9428..9458
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 9466..9482
/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 9490..9506
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 9527..9545
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"