Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012227 | pTALETF_v2 (HD) | 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:
- pTALETF_v2 (HD)
- Antibiotic Resistance:
- Ampicillin, Chloramphenicol
- Length:
- 8901 bp
- Type:
- Tale ToolBox kit
- Replication origin:
- ori
- Copy Number:
- High copy number
- Cloning Method:
- Enzyme digestion and ligation
pTALETF_v2 (HD) 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.
pTALETF_v2 (HD) vector Sequence
LOCUS 40924_42634 8901 bp DNA circular SYN 13-JAN-2022
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8901)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 8901)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8901
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 86..674
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 944..960
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 1416..1432
/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)."
enhancer 1459..1838
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1839..2042
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 2087..2105
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 2876..2906
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 2960..3616
/label=CmR
/note="chloramphenicol acetyltransferase"
CDS 3961..4263
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
CDS 4915..4935
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 4960..5109
/codon_start=1
/product="tetrameric repeat of the minimal activation
domain of herpes simplex virus VP16 (Beerli et al., 1998)"
/label=tetrameric repeat of the minimal activation
dom
/note="VP64"
/translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
DLDML"
CDS 5131..5184
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=2A peptide from Thosea asigna virus capsid
protein
/note="T2A"
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 5185..5898
/label=EGFP
/note="enhanced GFP"
polyA_signal 5922..6033
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
promoter 6156..6485
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 6534..7556
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
polyA_signal 7689..7822
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
CDS complement(7866..8723)
/label=AmpR
/note="beta-lactamase"
promoter complement(8724..8815)
/gene="bla"
/label=bla promoter
/note="AmpR promoter"