Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000355 | pBABE-neo-hTERT | 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:
- pBABE-neo-hTERT
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8709 bp
- Type:
- Mammalian Expression, Retroviral
- Selection Marker:
- Neomycin (select with G418)
- Copy Number:
- High Copy
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- pBABE 5'
- 3' Primer:
- pBABE 3'
pBABE-neo-hTERT 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.
pBABE-neo-hTERT vector Sequence
LOCUS Exported 8709 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Retroviral expression of hTERT. Used to create immortalized cell
lines..
ACCESSION .
VERSION .
KEYWORDS pBABE-neo-hTERT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8709)
AUTHORS Counter CM, Hahn WC, Wei W, Caddle SD, Beijersbergen RL, Lansdorp
PM, Sedivy JM, Weinberg RA
TITLE Dissociation among in vitro telomerase activity, telomere
maintenance, and cellular immortalization.
JOURNAL Proc Natl Acad Sci U S A 1998 Dec 8;95(25):14723-8.
PUBMED 9843956
REFERENCE 2 (bases 1 to 8709)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl
Acad Sci U S A"; date: "1998-12-8"; volume: "95(25)"; pages:
"14723-8"
FEATURES Location/Qualifiers
source 1..8709
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 11..480
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 545..744
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 945..1361
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1291..1313
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1329..1345
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
CDS 1441..4839
/codon_start=1
/gene="TERT"
/product="human telomerase reverse transcriptase, the
catalytic subunit of telomerase"
/label=hTERT
/note="prevents telomere shortening and allows
postsenescent cells to proliferate beyond crisis (Counter
et al., 1998); also known as EST2"
/translation="MPRAPRCRAVRSLLRSHYREVLPLATFVRRLGPQGWRLVQRGDPA
AFRALVAQCLVCVPWDARPPPAAPSFRQVSCLKELVARVLQRLCERGAKNVLAFGFALL
DGARGGPPEAFTTSVRSYLPNTVTDALRGSGAWGLLLRRVGDDVLVHLLARCALFVLVA
PSCAYQVCGPPLYQLGAATQARPPPHASGPRRRLGCERAWNHSVREAGVPLGLPAPGAR
RRGGSASRSLPLPKRPRRGAAPEPERTPVGQGSWAHPGRTRGPSDRGFCVVSPARPAEE
ATSLEGALSGTRHSHPSVGRQHHAGPPSTSRPPRPWDTPCPPVYAETKHFLYSSGDKEQ
LRPSFLLSSLRPSLTGARRLVETIFLGSRPWMPGTPRRLPRLPQRYWQMRPLFLELLGN
HAQCPYGVLLKTHCPLRAAVTPAAGVCAREKPQGSVAAPEEEDTDPRRLVQLLRQHSSP
WQVYGFVRACLRRLVPPGLWGSRHNERRFLRNTKKFISLGKHAKLSLQELTWKMSVRGC
AWLRRSPGVGCVPAAEHRLREEILAKFLHWLMSVYVVELLRSFFYVTETTFQKNRLFFY
RKSVWSKLQSIGIRQHLKRVQLRELSEAEVRQHREARPALLTSRLRFIPKPDGLRPIVN
MDYVVGARTFRREKRAERLTSRVKALFSVLNYERARRPGLLGASVLGLDDIHRAWRTFV
LRVRAQDPPPELYFVKVDVTGAYDTIPQDRLTEVIASIIKPQNTYCVRRYAVVQKAAHG
HVRKAFKSHVSTLTDLQPYMRQFVAHLQETSPLRDAVVIEQSSSLNEASSGLFDVFLRF
MCHHAVRIRGKSYVQCQGIPQGSILSTLLCSLCYGDMENKLFAGIRRDGLLLRLVDDFL
LVTPHLTHAKTFLRTLVRGVPEYGCVVNLRKTVVNFPVEDEALGGTAFVQMPAHGLFPW
CGLLLDTRTLEVQSDYSSYARTSIRASLTFNRGFKAGRNMRRKLFGVLRLKCHSLFLDL
QVNSLQTVCTNIYKILLLQAYRFHACVLQLPFHQQVWKNPTFFLRVISDTASLCYSILK
AKNAGMSLGAKGAAGPLPSEAVQWLCHQAFLLKLTRHRVTYVPLLGSLRTAQTQLSRKL
PGTTLTALEAAANPALPSDFKTILD"
primer_bind complement(4851..4871)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4856..5185
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 5036..5171
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5098..5117
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 5542..6336
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(5596..5615)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 6206..6225
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 6859..6878
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
LTR 7140..7609
/note="long terminal repeat from Moloney murine leukemia
virus"
CDS complement(7724..8584)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 8347..8366
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8585..8689)
/gene="bla"
/label=AmpR promoter