Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000856 | TLCV2 | 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:
- TLCV2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 16733 bp
- Type:
- Mammalian Expression, Lentiviral, CRISPR ; Doxycyc
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Promoter:
- U6
- 5' Primer:
- LNCX [5'-AGCTCGTTTAGTGAACCGTCAGATC-3']
- 3' Primer:
- EF1a-R [5'CACGGCGACTACTGCACTTA-3']
TLCV2 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.
TLCV2 vector Sequence
LOCUS V000856 16733 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000856
VERSION V000856
KEYWORDS TLCV2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 16733)
AUTHORS Barger CJ, Branick C, Chee L, Karpf AR
TITLE Pan-Cancer Analyses Reveal Genomic Features of FOXM1 Overexpression
in Cancer.
JOURNAL Cancers (Basel). 2019 Feb 21;11(2). pii: cancers11020251. doi:
10.3390/cancers11020251.
PUBMED 30795624
REFERENCE 2 (bases 1 to 16733)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 16733)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cancers
(Basel)."; date: "2019-02-21"; pages: " 10.3390/cancers11020251"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..16733
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 14..32
/label="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 49..67
/gene="tetO"
/label="tet operator"
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 85..103
/gene="tetO"
/label="tet operator"
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 121..139
/gene="tetO"
/label="tet operator"
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 156..174
/gene="tetO"
/label="tet operator"
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 192..210
/gene="tetO"
/label="tet operator"
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
promoter 222..260
/label="minimal CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 257..281
/label="LNCX"
/note="Human CMV promoter, forward primer"
CDS 316..4419
/label="Cas9"
/note="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
CDS 4420..4467
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label="nucleoplasmin NLS"
/translation="KRPAATKKAGQAKKKK"
CDS 4468..4491
/label="FLAG"
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 4501..4554
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label="T2A"
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 4555..5268
/label="EGFP"
/note="enhanced GFP"
promoter 5282..5493
/label="EF-1-alpha core promoter"
/note="core promoter for human elongation factor
EF-1-alpha"
CDS 5515..6111
/label="PuroR"
/note="puromycin N-acetyltransferase"
CDS 6121..6177
/label="P2A"
/note="2A peptide from porcine teschovirus-1 polyprotein"
CDS 6178..6921
/label="rtTA-Advanced"
/note="improved tetracycline-controlled transactivator"
misc_feature 6945..7533
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 7605..7838
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 7870..8094
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 8140..8568
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 8582..8911
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
promoter 8959..9006
/label="EM7 promoter"
/note="synthetic bacterial promoter"
CDS 9025..9396
/label="BleoR"
/note="antibiotic-binding protein"
polyA_signal 9529..9662
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(9699..9715)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9699..9715)
/label="M13 Reverse"
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(9712..9734)
/label="M13/pUC Reverse"
/note="In lacZ gene"
protein_bind 9723..9739
/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(9747..9777)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(9792..9813)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(9930..9947)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(10101..10689)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(10863..11720)
/label="AmpR"
/note="beta-lactamase"
promoter complement(11721..11825)
/label="AmpR promoter"
primer_bind complement(11900..11919)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 12091..12470
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 12471..12673
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 12688..12868
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 12915..13040
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 13533..13766
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 13951..13995
/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 14144..14185
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 14293..14410
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 14461..14701
/label="U6 promoter"
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 16591..16666
/label="gRNA scaffold"
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind 16711..16729
/label="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"