LOCUS       40924_33642        6599 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pNUS-SPnHcH, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6599)
  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 6599)
  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 6599)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6599)
  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..6599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             661..678
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     CDS             715..732
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             1801..2823
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIGKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRLSTRPRAK
                     E"
     primer_bind     complement(3980..3996)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4004..4020)
                     /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(4028..4058)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4073..4094)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4382..4970)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5144..6001)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6002..6106)
                     /label=AmpR promoter
     primer_bind     6580..6596
                     /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 attcatggct cgagctcgtt tccttcagct tctgctggtg
      601 acgctgacgc tcctcagtac tgctgcactc cctgtcagcg cgcatatgag atctggtacc
      661 ctggtgccac gcggttccat ggcgatatcg gatcagcttg cggccgcact cgagcaccac
      721 caccaccacc actgagaatt ctgtattacg ccgttttaag agctccttcg acgccttttt
      781 gtccttcagc tgctccacgg tgactcgctt ctctctcttt ccacagtgtc tcttttttct
      841 tttcactctc tatacaaatg tgagcgacct ccttttctgt acaacggcct tccggcgtgt
      901 gcttttctca tcgacctctc ctcgcttctt gggcactcct tcatcgagca aacaagacga
      961 gggaggtgac gcgattggtg agaccactac gagtatggac ggagctgtct gataccccct
     1021 gttttttatc ttgtaccccg ctctgcagtg acggtagccc gctgctgtgc ttcggtatcg
     1081 ccgcttcata cgcttctctc ttttttcaac gtgcggcgct gattcaaagg ttacctcaac
     1141 gcgacccgcc gctgcatcct ttgttgctcc tcttgtcgca aacaaacaaa aaacgtgctg
     1201 tgctttttcc tttatcgtgt ctttttgcaa agtctagagt ttaccgacaa gaccagaata
     1261 gcgcttatgt gtgtgtgttt gtgtgtgggt gtgtgtgccc atgtgtgaca aagaggccag
     1321 tgctttctct aaagcaagag cgcgaggttg ctggctaagg actctaagtc cctcacagtg
     1381 atggcgtgtt gcattttcta aaccgtcaat ccaacacgtc ggtagcatat cggttttgta
     1441 ggctcttctg gataccttcc ccagtaagac cgccactgct tcgtgactga agcactgtaa
     1501 atatcactga aaccgtcgcg ttgctttgtc taattatcac cgcctttgcc agctatcgcc
     1561 tcaggctgcg ctctttgact aatccactca ccacttctct gccttcttcc tccggttgtt
     1621 gttgtgtcct gcgtgtacat ggcgcgtgtc cttttcgagc aaacagctgt cttttctttc
     1681 acgataacac actcatatta aacgcgagta ttactcatca gtcaacgtca cattccgctc
     1741 tgtccacttc gaccttacac ctctacttgt ccaactatct tccacttgtc aagcgtcgac
     1801 atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcgg aaagttcgac
     1861 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
     1921 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
     1981 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt
     2041 ggggagttca gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg
     2101 caagacctgc ctgaaaccga actgcccgct gttcttcagc cggtcgcgga ggccatggat
     2161 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
     2221 atcggtcaat acactacatg gcgtgatttc atttgcgcga ttgctgatcc ccatgtgtat
     2281 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag
     2341 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc
     2401 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg
     2461 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct
     2521 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg
     2581 cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
     2641 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
     2701 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc
     2761 tgtgtagaag tactcgccga tagtggaaac cgacgcctca gcactcgtcc gagggcaaag
     2821 gaatagggat ccagcaggcg gagaaagagg gaagtataag gcggacgcat aggtcggagt
     2881 atcgagaaaa agaggcagag atgggtggtg gcggagcgcc cctctctgcc cttctctgtt
     2941 ttactgttcc cgccacgtcg ccaactcctc tttttgtttg ctctaatgtc ggtgctatcc
     3001 tcttcctctt tttactctcc gtttttcttc cttccgtttt gtcttgttat caccgttatt
     3061 tttcttcttc tttcttagcc gatttgggtc tcctgcctac ggcagcggtg atgagtcgca
     3121 cttccgtccc cctttcctct ccgtaagtac tccctcgatg cctcaggcgc ttctattttg
     3181 cggcactgtg ctgaccacct cccacgtgtg cagtgagagc gccagagaca ttcaggagaa
     3241 gagggaagag gggaagtaaa taccaaagcg aggaagatgt ctttctcgct gcttctcatc
     3301 ctgttgaccg gtgtgcacgg cggtgtgtcc tctcggcttc cctcctcctt ccctcgctcc
     3361 cctttttctg tgttttttct ttctaacatg attgcgcctg ctctttttgc cccgcagcgt
     3421 cgcagtgggt gtcacctgcg accgccgtac gttttactat atacatattt gtttctgtgc
     3481 tgatgccatt tgtccgctac tccatatcgt gcgtgtttgt tttccagtgg tgacgccttc
     3541 cgcatcattt taatcaattc gaggcgaaaa gggtgatttg agtgttggag tgggcttttt
     3601 tatttatttg actccaaaca tcttttcttt tcccggtatc ttcgggtccg tcacgcaggt
     3661 cggtgggtgt gtccgagtgc gcttctttac tcacgtgccg ctgcgcacat acacagattt
     3721 taaaagcacg catacacgat ctgtgccttc aaaactaatc aacaaaaaca aaaatacaaa
     3781 aaagacaaga ataagaggtc agcaacgcac ccacggctcc tttcttcctg atccacgtcg
     3841 tggccgctgt acgctctcaa acacgctttg gcgctgatgc gtgcctttat acaaagaaca
     3901 aagaagaagc gaaaaacggg agcgtgttgc tttggggatg tatgcgtgcc acagttctgc
     3961 ggtacaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4021 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4081 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4141 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4201 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4261 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4321 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4381 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4441 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4501 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4561 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4621 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4681 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4741 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4801 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     4861 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4921 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4981 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5041 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5101 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5161 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5221 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5281 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5341 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5401 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5461 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5521 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5581 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5641 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5701 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5761 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5821 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5881 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5941 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6001 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6061 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6121 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     6181 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     6241 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     6301 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     6361 agcagattgt actgagagtg cacgatatgc ggtgtgaaat accgcacaga tgcgtaagga
     6421 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     6481 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     6541 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcc
//
