LOCUS       40924_33632        6500 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pNUS-HnH, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6500)
  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 6500)
  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 6500)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6500)
  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..6500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             568..585
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             595..612
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     CDS             1702..2724
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIGKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRLSTRPRAK
                     E"
     primer_bind     complement(3881..3897)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3905..3921)
                     /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(3929..3959)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3974..3995)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4283..4871)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5045..5902)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5903..6007)
                     /label=AmpR promoter
     primer_bind     6481..6497
                     /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 attcatgcat catcatcatc atcacagcag cggcctggtg
      601 ccgcgcggca gccatatgct cgaggatatc ggtaccgaat tctgtattac gccgttttaa
      661 gagctccttc gacgcctttt tgtccttcag ctgctccacg gtgactcgct tctctctctt
      721 tccacagtgt ctcttttttc ttttcactct ctatacaaat gtgagcgacc tccttttctg
      781 tacaacggcc ttccggcgtg tgcttttctc atcgacctct cctcgcttct tgggcactcc
      841 ttcatcgagc aaacaagacg agggaggtga cgcgattggt gagaccacta cgagtatgga
      901 cggagctgtc tgataccccc tgttttttat cttgtacccc gctctgcagt gacggtagcc
      961 cgctgctgtg cttcggtatc gccgcttcat acgcttctct cttttttcaa cgtgcggcgc
     1021 tgattcaaag gttacctcaa cgcgacccgc cgctgcatcc tttgttgctc ctcttgtcgc
     1081 aaacaaacaa aaaacgtgct gtgctttttc ctttatcgtg tctttttgca aagtctagag
     1141 tttaccgaca agaccagaat agcgcttatg tgtgtgtgtt tgtgtgtggg tgtgtgtgcc
     1201 catgtgtgac aaagaggcca gtgctttctc taaagcaaga gcgcgaggtt gctggctaag
     1261 gactctaagt ccctcacagt gatggcgtgt tgcattttct aaaccgtcaa tccaacacgt
     1321 cggtagcata tcggttttgt aggctcttct ggataccttc cccagtaaga ccgccactgc
     1381 ttcgtgactg aagcactgta aatatcactg aaaccgtcgc gttgctttgt ctaattatca
     1441 ccgcctttgc cagctatcgc ctcaggctgc gctctttgac taatccactc accacttctc
     1501 tgccttcttc ctccggttgt tgttgtgtcc tgcgtgtaca tggcgcgtgt ccttttcgag
     1561 caaacagctg tcttttcttt cacgataaca cactcatatt aaacgcgagt attactcatc
     1621 agtcaacgtc acattccgct ctgtccactt cgaccttaca cctctacttg tccaactatc
     1681 ttccacttgt caagcgtcga catgaaaaag cctgaactca ccgcgacgtc tgtcgagaag
     1741 tttctgatcg gaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     1801 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     1861 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
     1921 attccggaag tgcttgacat tggggagttc agcgagagcc tgacctattg catctcccgc
     1981 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttcttcag
     2041 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
     2101 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catttgcgcg
     2161 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
     2221 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
     2281 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
     2341 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc
     2401 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
     2461 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa
     2521 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
     2581 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
     2641 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcctc
     2701 agcactcgtc cgagggcaaa ggaataggga tccagcaggc ggagaaagag ggaagtataa
     2761 ggcggacgca taggtcggag tatcgagaaa aagaggcaga gatgggtggt ggcggagcgc
     2821 ccctctctgc ccttctctgt tttactgttc ccgccacgtc gccaactcct ctttttgttt
     2881 gctctaatgt cggtgctatc ctcttcctct ttttactctc cgtttttctt ccttccgttt
     2941 tgtcttgtta tcaccgttat ttttcttctt ctttcttagc cgatttgggt ctcctgccta
     3001 cggcagcggt gatgagtcgc acttccgtcc ccctttcctc tccgtaagta ctccctcgat
     3061 gcctcaggcg cttctatttt gcggcactgt gctgaccacc tcccacgtgt gcagtgagag
     3121 cgccagagac attcaggaga agagggaaga ggggaagtaa ataccaaagc gaggaagatg
     3181 tctttctcgc tgcttctcat cctgttgacc ggtgtgcacg gcggtgtgtc ctctcggctt
     3241 ccctcctcct tccctcgctc ccctttttct gtgttttttc tttctaacat gattgcgcct
     3301 gctctttttg ccccgcagcg tcgcagtggg tgtcacctgc gaccgccgta cgttttacta
     3361 tatacatatt tgtttctgtg ctgatgccat ttgtccgcta ctccatatcg tgcgtgtttg
     3421 ttttccagtg gtgacgcctt ccgcatcatt ttaatcaatt cgaggcgaaa agggtgattt
     3481 gagtgttgga gtgggctttt ttatttattt gactccaaac atcttttctt ttcccggtat
     3541 cttcgggtcc gtcacgcagg tcggtgggtg tgtccgagtg cgcttcttta ctcacgtgcc
     3601 gctgcgcaca tacacagatt ttaaaagcac gcatacacga tctgtgcctt caaaactaat
     3661 caacaaaaac aaaaatacaa aaaagacaag aataagaggt cagcaacgca cccacggctc
     3721 ctttcttcct gatccacgtc gtggccgctg tacgctctca aacacgcttt ggcgctgatg
     3781 cgtgccttta tacaaagaac aaagaagaag cgaaaaacgg gagcgtgttg ctttggggat
     3841 gtatgcgtgc cacagttctg cggtacaatt cgtaatcatg gtcatagctg tttcctgtgt
     3901 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3961 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4021 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4081 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4141 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4201 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4261 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4321 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4381 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4441 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     4501 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4561 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4621 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4681 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     4741 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4801 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4861 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4921 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4981 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5041 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5101 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5161 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5221 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5281 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5341 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5401 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5461 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5521 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5581 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5641 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5701 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5761 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5821 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5881 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5941 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6001 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     6061 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     6121 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     6181 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     6241 ggcttaacta tgcggcatca gagcagattg tactgagagt gcacgatatg cggtgtgaaa
     6301 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
     6361 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     6421 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     6481 gtaaaacgac ggccagtgcc
//
