Basic Vector Information
pTrypRNAiGate 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 ℃.
pTrypRNAiGate vector Sequence
LOCUS 40924_44304 9177 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pTrypRNAiGate, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9177)
AUTHORS Kalidas S, Li Q, Phillips MA.
TITLE A Gateway((R)) compatible vector for gene silencing in bloodstream
form Trypanosoma brucei
JOURNAL Mol. Biochem. Parasitol. 178 (1-2), 51-55 (2011)
PUBMED 21420443
REFERENCE 2 (bases 1 to 9177)
AUTHORS Kalidas S, Li Q, Phillips M.
TITLE Direct Submission
JOURNAL Submitted (30-MAR-2011) Department of Pharmacology, UT Southwestern
Medical Center, 6001 Forest Park Rd, Dallas, TX 75390-9041, USA
REFERENCE 3 (bases 1 to 9177)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9177)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol.
Biochem. Parasitol. 178 (1-2), 51-55 (2011)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(30-MAR-2011) Department of Pharmacology, UT Southwestern Medical
Center, 6001 Forest Park Rd, Dallas, TX 75390-9041, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9177
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature complement(100..732)
/label=rRNA locus intergenic fragment
/note="rRNA locus intergenic fragment"
CDS complement(1085..1456)
/codon_start=1
/label=BleoR
/note="antibiotic-binding protein"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
promoter complement(1600..1618)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1883..1901
/label=tet operator
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 1904..1922
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2053..2177
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 2202..2232
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 2286..2942
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 3287..3589
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(3633..3757)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
protein_bind 4213..4337
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
CDS complement(4381..4683)
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
CDS complement(5028..5684)
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(5738..5768)
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
protein_bind complement(5793..5917)
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter complement(6413..6431)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
promoter 7217..7321
/label=AmpR promoter
CDS 7322..8179
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHSMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 8353..8941
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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