pNUS-GFPcH vector (V004256)

MSDS

Basic Vector Information

Vector Name:: pNUS-GFPcH

Antibiotic Resistance:: Ampicillin

Length:: 7190 bp

Type:: Expression vector

Replication origin:: ori

Source/Author:: Tetaud E, Lecuix I, Sheldrake T, Baltz T, Fairlamb AH.

pNUS-GFPcH 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-GFPcH vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_33622        7190 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pNUS-GFPcH, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7190)
  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 7190)
  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 7190)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7190)
  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..7190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             583..1296
                     /codon_start=1
                     /label=GFP (S65T)
                     /note="S65T variant of Aequorea victoria green fluorescent 
                     protein (Heim et al., 1995)"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1306..1323
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             2392..3414
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIGKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRLSTRPRAK
                     E"
     primer_bind     complement(4571..4587)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4595..4611)
                     /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(4619..4649)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4664..4685)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4973..5561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5735..6592)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6593..6697)
                     /label=AmpR promoter
     primer_bind     7171..7187
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"

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