LOCUS       Exported                7190 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pNUS-GFPcH, complete sequence.
ACCESSION   AY056838
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7190
                     /organism="Expression vector pNUS-GFPcH"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:174819"
     CDS             583..1299
                     /codon_start=1
                     /gene="GFP"
                     /product="green fluorescent protein"
                     /label=GFP
                     /protein_id="AAL17988.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            583..1299
                     /gene="GFP"
                     /label=GFP
     CDS             1306..1323
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2392..3417
                     /codon_start=1
                     /gene="hygR"
                     /product="hygromycin phosphotransferase"
                     /label=hygR
                     /note="provides resistance to hygromycin"
                     /protein_id="AAL17987.1"
                     /translation="MKKPELTATSVEKFLIGKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRLSTRPRAK
                     E"
     gene            2392..3417
                     /gene="hygR"
                     /label=hygR
     primer_bind     complement(4571..4587)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4595..4611
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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    4664..4685
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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(5732..6592)
                     /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"
     promoter        complement(6593..6697)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7171..7187
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"