Basic Vector Information
- Vector Name:
- TRMPV-ns
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7838 bp
- Type:
- Retroviral Tet-shRNA expression vector
- Replication origin:
- ori
- Source/Author:
- Zuber J, McJunkin K, Fellmann C, Dow LE, Taylor MJ, Hannon GJ, Lowe SW.
- Promoter:
- mPGK
TRMPV-ns vector Vector Map
Plasmid Resuspension 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 ℃.
TRMPV-ns vector Sequence
LOCUS 40924_48967 7838 bp DNA circular SYN 18-DEC-2018
DEFINITION Retroviral Tet-shRNA expression vector TRMPV-ns, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7838)
AUTHORS Zuber J, McJunkin K, Fellmann C, Dow LE, Taylor MJ, Hannon GJ, Lowe
SW.
TITLE Toolkit for evaluating genes required for proliferation and survival
using tetracycline-regulated RNAi
JOURNAL Nat. Biotechnol. (2010) In press
PUBMED 21131983
REFERENCE 2 (bases 1 to 7838)
AUTHORS Zuber J, McJunkin K, Fellmann C, Dow LE, Taylor MJ, Hannon GJ, Lowe
SW.
TITLE Direct Submission
JOURNAL Submitted (01-NOV-2010) Scott Lowe Lab, Cold Spring Harbor
Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USA
REFERENCE 3 (bases 1 to 7838)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7838)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat.
Biotechnol. (2010) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(01-NOV-2010) Scott Lowe Lab, Cold Spring Harbor Laboratory, 1
Bungtown Road, Cold Spring Harbor, NY 11724, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7838
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 5..309
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 310..513
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 514..689
/label=5' LTR (truncated)
/note="truncated long terminal repeat from Moloney murine
sarcoma virus"
misc_feature 752..951
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 1152..1568
/codon_start=1
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
/translation="GQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTF
NVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKP
PPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
protein_bind 1618..1888
/label=tetracycline response element
/note="contains seven copies of the tetracycline operator
tetO"
promoter 1921..1959
/label=minimal CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 2058..2732
/codon_start=1
/label=DsRed2
/note="improved tetrameric variant of DsRed fluorescent
protein"
/translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
LFL"
ncRNA 2804..2925
/label=5' miR-30a
/note="sequence upstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
ncRNA 2953..2967
/label=miR-30a loop
/note="loop from the the 71-nt precursor of the human
miR-30a microRNA (Zeng et al., 2002)"
/ncRNA_class="miRNA"
ncRNA 2995..3121
/label=3' miR-30a
/note="sequence downstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
promoter 3144..3643
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 3666..4382
/codon_start=1
/label=Venus
/note="yellow fluorescent protein (YFP) with fast and
efficient maturation (Nagai et al., 2002)"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
LTR 4544..4969
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
Moloney murine leukemia virus"
promoter 5235..5564
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin complement(5857..6445)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6619..7476)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(7477..7581)
/label=AmpR promoter
enhancer 7767..7838
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
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