LOCUS       40924_34600        3551 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pPICZalphaH6E, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3551)
  AUTHORS   Janakiraman VN, Cabanne C, Dieryck W, Hocquellet A, Joucla G, Le 
            Senechal C, Chaignepain S, Costaglioli P, Santarelli X, Garbay B, 
            Noubhani A.
  TITLE     Production and purification of recombinant human hepcidin-25 with 
            authentic N and C-termini
  JOURNAL   J. Biotechnol. 195, 89-92 (2015)
  PUBMED    25562424
REFERENCE   2  (bases 1 to 3551)
  AUTHORS   Dieryck W, Narasimhan Janakiraman V, Cabanne C, Hocquellet A, Joucla
            G, Le Senechal C, Chaignepain S, Costaglioli P, Santarelli X, Garbay
            B, Noubhani A.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-JUN-2014) ENSTBB, Institut Polytechnique de Bordeaux, 
            146 rue Leo Saignat, Bordeaux, gironde 33076, France
REFERENCE   3  (bases 1 to 3551)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3551)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Biotechnol. 195, 89-92 (2015)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-JUN-2014) ENSTBB, Institut Polytechnique de Bordeaux, 146 rue 
            Leo Saignat, Bordeaux, gironde 33076, France"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..940
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             941..1207
                     /codon_start=1
                     /label=alpha-factor secretion signal
                     /note="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
     CDS             1238..1255
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             1301..1315
                     /codon_start=1
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
                     /translation="DDDDK"
     misc_feature    1315..1359
                     /label=multiple cloning site
                     /note="multiple cloning site"
     terminator      1375..1621
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1636..2047
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2055..2102
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             2121..2492
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      2561..2808
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(2883..3471)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag
       61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt
      121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc
      181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta
      241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta
      301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg
      361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct
      421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg
      481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt
      541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct
      601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct
      661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact
      721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat
      781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
      841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
      901 caacttgaga agatcaaaaa acaactaatt attcgaaacg atgagatttc cttcaatttt
      961 tactgctgtt ttattcgcag catcctccgc attagctgct ccagtcaaca ctacaacaga
     1021 agatgaaacg gcacaaattc cggctgaagc tgtcatcggt tactcagatt tagaagggga
     1081 tttcgatgtt gctgttttgc cattttccaa cagcacaaat aacgggttat tgtttataaa
     1141 tactactatt gccagcattg ctgctaaaga agaaggggta tctctcgaga aaagagaggc
     1201 tgaagctgaa ttgtggtcac catcgtatta cgcgtctcat caccatcacc atcactcgag
     1261 ctcgaacaac aacaacaata acaataacaa caacctcggg gatgacgatg acaaggtacc
     1321 ggaattcgca tgctctagag tcgacctgca gtgaccggtc ttgctagatt ctaatcaaga
     1381 ggatgtcaga atgccatttg cctgagagat gcaggcttca tttttgatac ttttttattt
     1441 gtaacctata tagtatagga ttttttttgt cattttgttt cttctcgtac gagcttgctc
     1501 ctgatcagcc tatctcgcag ctgatgaata tcttgtggta ggggtttggg aaaatcattc
     1561 gagtttgatg tttttcttgg tatttcccac tcctcttcag agtacagaag attaagtgag
     1621 accttcgttt gtgcggatcc cccacacacc atagcttcaa aatgtttcta ctcctttttt
     1681 actcttccag attttctcgg actccgcgca tcgccgtacc acttcaaaac acccaagcac
     1741 agcatactaa attttccctc tttcttcctc tagggtgtcg ttaattaccc gtactaaagg
     1801 tttggaaaag aaaaaagaga ccgcctcgtt tctttttctt cgtcgaaaaa ggcaataaaa
     1861 atttttatca cgtttctttt tcttgaaatt ttttttttta gtttttttct ctttcagtga
     1921 cctccattga tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt
     1981 tcttgttcta ttacaacttt ttttacttct tgttcattag aaagaaagca tagcaatcta
     2041 atctaagggg cggtgttgac aattaatcat cggcatagta tatcggcata gtataatacg
     2101 acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc
     2161 gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg
     2221 tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc
     2281 aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc
     2341 tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca
     2401 tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca
     2461 actgcgtgca cttcgtggcc gaggagcagg actgacacgt ccgacggcgg cccacgggtc
     2521 ccaggcctcg gagatccgtc ccccttttcc tttgtcgata tcatgtaatt agttatgtca
     2581 cgcttacatt cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa
     2641 cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt
     2701 atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg
     2761 aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taatttgcaa gctggagacc
     2821 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2881 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2941 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3001 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3061 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3121 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3181 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3241 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3301 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     3361 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3421 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3481 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3541 gtcatgagat c
//
