LOCUS       pFLD.        4409 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Pichia pastoris vector for methylamine-inducible intracellular 
            expression of a protein.
ACCESSION   .
VERSION     .
KEYWORDS    pFLD
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4409)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4409)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Linearize within the FLD1 promoter for integration.
FEATURES             Location/Qualifiers
     source          1..4409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..597
                     /label=FLD1 promoter
                     /note="methanol- and methylamine-inducible promoter for
                     Pichia pastoris formaldehyde dehydrogenase"
     misc_feature    598..674
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             679..720
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
     CDS             730..747
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      827..1073
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1440..1544
                     /label=AmpR promoter
     CDS             1545..2402
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2576..3164
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        3218..3625
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3632..3679
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             3698..4069
                     /label=BleoR
                     /note="antibiotic-binding protein"
     terminator      4138..4385
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
ORIGIN
        1 gcatgcagga atctctggca cggtgctaat ggtagttatc caacggagct gaggtagtcg
       61 atatatctgg atatgccgcc tataggataa aaacaggaga gggtgaacct tgcttatggc
      121 tactagattg ttcttgtact ctgaattctc attatgggaa actaaactaa tctcatctgt
      181 gtgttgcagt actattgaat cgttgtagta tctacctgga gggcattcca tgaattagtg
      241 agataacaga gttgggtaac tagagagaat aatagacgta tgcatgatta ctacacaacg
      301 gatgtcgcac tctttcctta gttaaaacta tcatccaatc acaagatgcg ggctggaaag
      361 acttgctccc gaaggataat cttctgcttc tatctccctt cctcatatgg tttcgcaggg
      421 ctcatgcccc ttcttccttc gaactgcccg atgaggaagt ccttagccta tcaaagaatt
      481 cgggaccatc atcgattttt agagccttac ctgatcgcaa tcaggatttc actactcata
      541 taaatacatc gctcaaagct ccaactttgc ttgttcatac aattcttgat attcacacaa
      601 ttgttcacgt ggcccagccg gccgtctcgg atcggtacct cgagccgcgg cggccgccag
      661 cttgggccca attggggtgg taagcctatc cctaaccctc tcctcggtct cgattctacg
      721 ggtgtcgacc atcatcatca tcatcattga gtttgtagcc ttagacatga ctgttcctca
      781 gttcaagttg ggcacttacg agaagaccgg tcttgctaga ttctaatcaa gaggatgtca
      841 gaatgccatt tgcctgagag atgcaggctt catttttgat acttttttat ttgtaaccta
      901 tatagtatag gatttttttt gtcattttgt ttcttctcgt acgagcttgc tcctgatcag
      961 cctatctcgc agctgatgaa tatcttgtgg taggggtttg ggaaaatcat tcgagtttga
     1021 tgtttttctt ggtatttccc actcctcttc agagtacaga agattaagtg agaccttcgt
     1081 ttgtgcggat ccggcgcgcc gatccgcctg atgcggtatt ttctccttac gcatctgtgc
     1141 ggtatttcac accgcatata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     1201 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     1261 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     1321 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     1381 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     1441 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1501 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1561 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     1621 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     1681 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     1741 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     1801 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     1861 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     1921 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     1981 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2041 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2101 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2161 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2221 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2281 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2341 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2401 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     2461 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     2521 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2581 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2641 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2701 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2761 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2821 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2881 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2941 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3001 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3061 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3121 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3181 cctttttacg gttcctggcc ttttgctggc cttttgctca cacaccatag cttcaaaatg
     3241 tttctactcc ttttttactc ttccagattt tctcggactc cgcgcatcgc cgtaccactt
     3301 caaaacaccc aagcacagca tactaaattt cccctctttc ttcctctagg gtgtcgttaa
     3361 ttacccgtac taaaggtttg gaaaagaaaa aagagaccgc ctcgtttctt tttcttcgtc
     3421 gaaaaaggca ataaaaattt ttatcacgtt tctttttctt gaaaattttt ttttttgatt
     3481 tttttctctt tcgatgacct cccattgata tttaagttaa taaacggtct tcaatttctc
     3541 aagtttcagt ttcatttttc ttgttctatt acaacttttt ttacttcttg ctcattagaa
     3601 agaaagcata gcaatctaat ctaagggcgg cgttgacaat taatcatcgg catagtatat
     3661 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
     3721 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
     3781 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
     3841 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
     3901 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
     3961 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcagg agttcgccct
     4021 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtccg
     4081 acgcggcccg acgggtccga ggcctcggag atccgtcccc cttttccttt gtcgatatca
     4141 tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa
     4201 aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata gttatgttag
     4261 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg
     4321 catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa
     4381 tttgcaagct gaattgcggc cgcagatct
//
