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"
ORIGIN
        1 aagcttaccg acaagaccag aatagcgctt atgtgtgtgt gtttgtgtgt gggtgtgtgt
       61 gcccatgtgt gacaaagagg ccagtgcttt ctctaaagca agagcgcgag gttgctggct
      121 aaggactcta agtccctcac agtgatggcg tgttgcattt tctaaaccgt caatccaaca
      181 cgtcggtagc atatcggttt tgtaggctct tctggatacc ttccccagta agaccgccac
      241 tgcttcgtga ctgaagcact gtaaatatca ctgaaaccgt cgcgttgctt tgtctaatta
      301 tcaccgcctt tgccagctat cgcctcaggc tgcgctcttt gactaatcca ctcaccactt
      361 ctctgccttc ttcctccggt tgttgttgtg tcctgcgtgt acatggcgcg tgtccttttc
      421 gagcaaacag ctgtcttttc tttcacgata acacactcat attaaacgcg agtattactc
      481 atcagtcaac gtcacattcc gctctgtcca cttcgacctt acacctctac ttgtccaact
      541 atcttccact tgtcaagcga attccatatg agatctggta ccatgagcaa gggcgaggaa
      601 ctgttcactg gcgtggtccc aattctcgtg gaactggatg gcgatgtgaa tgggcacaaa
      661 ttttctgtca gcggagaggg tgaaggtgat gccacatacg gaaagctcac cctgaaattc
      721 atctgcacca ctggaaagct ccctgtgcca tggccaacac tggtcactac cttcacctat
      781 ggcgtgcagt gcttttccag atacccagac cacatgaagc agcatgactt tttcaagagc
      841 gccatgcccg agggctatgt gcaggagaga accatctttt tcaaagatga cgggaactac
      901 aagacccgcg ctgaagtcaa gttcgaaggt gacaccctgg tgaatagaat cgagctgaag
      961 ggcattgact ttaaggagga tggaaacatt ctcggccaca agctggaata caactataac
     1021 tcccacaatg tgtacatcat ggccgacaag caaaagaatg gcatcaaggt caacttcaag
     1081 atcagacaca acattgagga tggatccgtg cagctggccg accattatca acagaacact
     1141 ccaatcggcg acggccctgt gctcctccca gacaaccatt acctgtccac ccagtctgcc
     1201 ctgtctaaag atcccaacga aaagagagac cacatggtcc tgctggagtt tgtgaccgct
     1261 gctgggatca cacatggcat ggacgagctg tacaagtgac tcgagcacca ccaccaccac
     1321 cactgagaat tctgtattac gccgttttaa gagctccttc gacgcctttt tgtccttcag
     1381 ctgctccacg gtgactcgct tctctctctt tccacagtgt ctcttttttc ttttcactct
     1441 ctatacaaat gtgagcgacc tccttttctg tacaacggcc ttccggcgtg tgcttttctc
     1501 atcgacctct cctcgcttct tgggcactcc ttcatcgagc aaacaagacg agggaggtga
     1561 cgcgattggt gagaccacta cgagtatgga cggagctgtc tgataccccc tgttttttat
     1621 cttgtacccc gctctgcagt gacggtagcc cgctgctgtg cttcggtatc gccgcttcat
     1681 acgcttctct cttttttcaa cgtgcggcgc tgattcaaag gttacctcaa cgcgacccgc
     1741 cgctgcatcc tttgttgctc ctcttgtcgc aaacaaacaa aaaacgtgct gtgctttttc
     1801 ctttatcgtg tctttttgca aagtctagag tttaccgaca agaccagaat agcgcttatg
     1861 tgtgtgtgtt tgtgtgtggg tgtgtgtgcc catgtgtgac aaagaggcca gtgctttctc
     1921 taaagcaaga gcgcgaggtt gctggctaag gactctaagt ccctcacagt gatggcgtgt
     1981 tgcattttct aaaccgtcaa tccaacacgt cggtagcata tcggttttgt aggctcttct
     2041 ggataccttc cccagtaaga ccgccactgc ttcgtgactg aagcactgta aatatcactg
     2101 aaaccgtcgc gttgctttgt ctaattatca ccgcctttgc cagctatcgc ctcaggctgc
     2161 gctctttgac taatccactc accacttctc tgccttcttc ctccggttgt tgttgtgtcc
     2221 tgcgtgtaca tggcgcgtgt ccttttcgag caaacagctg tcttttcttt cacgataaca
     2281 cactcatatt aaacgcgagt attactcatc agtcaacgtc acattccgct ctgtccactt
     2341 cgaccttaca cctctacttg tccaactatc ttccacttgt caagcgtcga catgaaaaag
     2401 cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg gaaagttcga cagcgtctcc
     2461 gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg
     2521 cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt
     2581 tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggagttc
     2641 agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg
     2701 cctgaaaccg aactgcccgc tgttcttcag ccggtcgcgg aggccatgga tgcgatcgct
     2761 gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa
     2821 tacactacat ggcgtgattt catttgcgcg attgctgatc cccatgtgta tcactggcaa
     2881 actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt
     2941 tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat
     3001 gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg
     3061 gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag
     3121 cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg
     3181 gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc
     3241 gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact
     3301 gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa
     3361 gtactcgccg atagtggaaa ccgacgcctc agcactcgtc cgagggcaaa ggaataggga
     3421 tccagcaggc ggagaaagag ggaagtataa ggcggacgca taggtcggag tatcgagaaa
     3481 aagaggcaga gatgggtggt ggcggagcgc ccctctctgc ccttctctgt tttactgttc
     3541 ccgccacgtc gccaactcct ctttttgttt gctctaatgt cggtgctatc ctcttcctct
     3601 ttttactctc cgtttttctt ccttccgttt tgtcttgtta tcaccgttat ttttcttctt
     3661 ctttcttagc cgatttgggt ctcctgccta cggcagcggt gatgagtcgc acttccgtcc
     3721 ccctttcctc tccgtaagta ctccctcgat gcctcaggcg cttctatttt gcggcactgt
     3781 gctgaccacc tcccacgtgt gcagtgagag cgccagagac attcaggaga agagggaaga
     3841 ggggaagtaa ataccaaagc gaggaagatg tctttctcgc tgcttctcat cctgttgacc
     3901 ggtgtgcacg gcggtgtgtc ctctcggctt ccctcctcct tccctcgctc ccctttttct
     3961 gtgttttttc tttctaacat gattgcgcct gctctttttg ccccgcagcg tcgcagtggg
     4021 tgtcacctgc gaccgccgta cgttttacta tatacatatt tgtttctgtg ctgatgccat
     4081 ttgtccgcta ctccatatcg tgcgtgtttg ttttccagtg gtgacgcctt ccgcatcatt
     4141 ttaatcaatt cgaggcgaaa agggtgattt gagtgttgga gtgggctttt ttatttattt
     4201 gactccaaac atcttttctt ttcccggtat cttcgggtcc gtcacgcagg tcggtgggtg
     4261 tgtccgagtg cgcttcttta ctcacgtgcc gctgcgcaca tacacagatt ttaaaagcac
     4321 gcatacacga tctgtgcctt caaaactaat caacaaaaac aaaaatacaa aaaagacaag
     4381 aataagaggt cagcaacgca cccacggctc ctttcttcct gatccacgtc gtggccgctg
     4441 tacgctctca aacacgcttt ggcgctgatg cgtgccttta tacaaagaac aaagaagaag
     4501 cgaaaaacgg gagcgtgttg ctttggggat gtatgcgtgc cacagttctg cggtacaatt
     4561 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4621 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4681 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4741 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4801 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4861 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4921 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4981 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5041 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5101 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5161 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5221 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5281 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5341 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5401 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5461 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5521 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5581 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5641 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5701 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5761 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5821 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5881 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5941 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6001 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6061 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6121 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6181 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6241 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6301 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6361 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6421 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6481 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6541 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6601 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6661 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6721 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     6781 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     6841 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     6901 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     6961 tactgagagt gcacgatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     7021 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     7081 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     7141 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
//
