Basic Vector Information
- Vector Name:
- pNUS-HcH
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6521 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Tetaud E, Lecuix I, Sheldrake T, Baltz T, Fairlamb AH.
pNUS-HcH 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 ℃.
pNUS-HcH vector Sequence
LOCUS 40924_33627 6521 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pNUS-HcH, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6521)
AUTHORS Tetaud E, Lecuix I, Sheldrake T, Baltz T, Fairlamb AH.
TITLE A new expression vector for Crithidia fasciculata and Leishmania
JOURNAL Mol. Biochem. Parasitol. 120 (2), 195-204 (2002)
PUBMED 11897125
REFERENCE 2 (bases 1 to 6521)
AUTHORS Tetaud E, Leciux I, Sheldrake T, Baltz T, Fairlamb AH.
TITLE Direct Submission
JOURNAL Submitted (24-SEP-2001) Parasitologie Moleculaire, UMR CNRS 5016,
146 Rue Leo Saignat, Bordeaux 33076, France
REFERENCE 3 (bases 1 to 6521)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6521)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol.
Biochem. Parasitol."; date: "2002"; volume: "120"; issue: "2";
pages: "195-204"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(24-SEP-2001) Parasitologie Moleculaire, UMR CNRS 5016, 146 Rue Leo
Saignat, Bordeaux 33076, France"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6521
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 583..600
/codon_start=1
/label=thrombin site
/note="thrombin recognition and cleavage site"
/translation="LVPRGS"
CDS 637..654
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
CDS 1723..2745
/codon_start=1
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
/translation="MKKPELTATSVEKFLIGKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRLSTRPRAK
E"
primer_bind complement(3902..3918)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3926..3942)
/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)."
promoter complement(3950..3980)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3995..4016)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4304..4892)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5066..5923)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5924..6028)
/label=AmpR promoter
primer_bind 6502..6518
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
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