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"
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 ccctggtgcc acgcggttcc
      601 atggcgatat cggatcagct tgcggccgca ctcgagcacc accaccacca ccactgagaa
      661 ttctgtatta cgccgtttta agagctcctt cgacgccttt ttgtccttca gctgctccac
      721 ggtgactcgc ttctctctct ttccacagtg tctctttttt cttttcactc tctatacaaa
      781 tgtgagcgac ctccttttct gtacaacggc cttccggcgt gtgcttttct catcgacctc
      841 tcctcgcttc ttgggcactc cttcatcgag caaacaagac gagggaggtg acgcgattgg
      901 tgagaccact acgagtatgg acggagctgt ctgatacccc ctgtttttta tcttgtaccc
      961 cgctctgcag tgacggtagc ccgctgctgt gcttcggtat cgccgcttca tacgcttctc
     1021 tcttttttca acgtgcggcg ctgattcaaa ggttacctca acgcgacccg ccgctgcatc
     1081 ctttgttgct cctcttgtcg caaacaaaca aaaaacgtgc tgtgcttttt cctttatcgt
     1141 gtctttttgc aaagtctaga gtttaccgac aagaccagaa tagcgcttat gtgtgtgtgt
     1201 ttgtgtgtgg gtgtgtgtgc ccatgtgtga caaagaggcc agtgctttct ctaaagcaag
     1261 agcgcgaggt tgctggctaa ggactctaag tccctcacag tgatggcgtg ttgcattttc
     1321 taaaccgtca atccaacacg tcggtagcat atcggttttg taggctcttc tggatacctt
     1381 ccccagtaag accgccactg cttcgtgact gaagcactgt aaatatcact gaaaccgtcg
     1441 cgttgctttg tctaattatc accgcctttg ccagctatcg cctcaggctg cgctctttga
     1501 ctaatccact caccacttct ctgccttctt cctccggttg ttgttgtgtc ctgcgtgtac
     1561 atggcgcgtg tccttttcga gcaaacagct gtcttttctt tcacgataac acactcatat
     1621 taaacgcgag tattactcat cagtcaacgt cacattccgc tctgtccact tcgaccttac
     1681 acctctactt gtccaactat cttccacttg tcaagcgtcg acatgaaaaa gcctgaactc
     1741 accgcgacgt ctgtcgagaa gtttctgatc ggaaagttcg acagcgtctc cgacctgatg
     1801 cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat
     1861 gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac
     1921 tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggagtt cagcgagagc
     1981 ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc
     2041 gaactgcccg ctgttcttca gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat
     2101 cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca
     2161 tggcgtgatt tcatttgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg
     2221 gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag
     2281 gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg
     2341 gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa
     2401 tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg
     2461 cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg ggcgtatatg
     2521 ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca
     2581 gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt
     2641 acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc
     2701 gatagtggaa accgacgcct cagcactcgt ccgagggcaa aggaataggg atccagcagg
     2761 cggagaaaga gggaagtata aggcggacgc ataggtcgga gtatcgagaa aaagaggcag
     2821 agatgggtgg tggcggagcg cccctctctg cccttctctg ttttactgtt cccgccacgt
     2881 cgccaactcc tctttttgtt tgctctaatg tcggtgctat cctcttcctc tttttactct
     2941 ccgtttttct tccttccgtt ttgtcttgtt atcaccgtta tttttcttct tctttcttag
     3001 ccgatttggg tctcctgcct acggcagcgg tgatgagtcg cacttccgtc cccctttcct
     3061 ctccgtaagt actccctcga tgcctcaggc gcttctattt tgcggcactg tgctgaccac
     3121 ctcccacgtg tgcagtgaga gcgccagaga cattcaggag aagagggaag aggggaagta
     3181 aataccaaag cgaggaagat gtctttctcg ctgcttctca tcctgttgac cggtgtgcac
     3241 ggcggtgtgt cctctcggct tccctcctcc ttccctcgct cccctttttc tgtgtttttt
     3301 ctttctaaca tgattgcgcc tgctcttttt gccccgcagc gtcgcagtgg gtgtcacctg
     3361 cgaccgccgt acgttttact atatacatat ttgtttctgt gctgatgcca tttgtccgct
     3421 actccatatc gtgcgtgttt gttttccagt ggtgacgcct tccgcatcat tttaatcaat
     3481 tcgaggcgaa aagggtgatt tgagtgttgg agtgggcttt tttatttatt tgactccaaa
     3541 catcttttct tttcccggta tcttcgggtc cgtcacgcag gtcggtgggt gtgtccgagt
     3601 gcgcttcttt actcacgtgc cgctgcgcac atacacagat tttaaaagca cgcatacacg
     3661 atctgtgcct tcaaaactaa tcaacaaaaa caaaaataca aaaaagacaa gaataagagg
     3721 tcagcaacgc acccacggct cctttcttcc tgatccacgt cgtggccgct gtacgctctc
     3781 aaacacgctt tggcgctgat gcgtgccttt atacaaagaa caaagaagaa gcgaaaaacg
     3841 ggagcgtgtt gctttgggga tgtatgcgtg ccacagttct gcggtacaat tcgtaatcat
     3901 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3961 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4021 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4081 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4141 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4201 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4261 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4321 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4381 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4441 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     4501 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4561 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4621 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4681 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4741 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4801 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4861 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4921 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4981 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5041 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5101 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5161 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5221 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5281 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5341 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5401 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5461 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5521 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5581 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5641 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5701 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5761 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5821 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5881 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5941 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6001 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     6061 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     6121 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     6181 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     6241 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     6301 tgcacgatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     6361 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     6421 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     6481 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc c
//
