LOCUS       pFLD(alpha).        4694 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Pichia pastoris vector for methylamine-inducible expression of a 
            secreted protein.
ACCESSION   .
VERSION     .
KEYWORDS    pFLD(alpha)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4694)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4694)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Linearize within the FLD1 promoter for integration.
FEATURES             Location/Qualifiers
     source          1..4694
                     /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"
     CDS             622..888
                     /label=alpha-factor secretion signal
                     /note="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
     misc_feature    889..959
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             964..1005
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
     CDS             1015..1032
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      1112..1358
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1725..1829
                     /label=AmpR promoter
     CDS             1830..2687
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2861..3449
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        3503..3910
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3917..3964
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             3983..4354
                     /label=BleoR
                     /note="antibiotic-binding protein"
     terminator      4423..4670
                     /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 ttcaacaact atttcgaaac gatgagattt ccttcaattt ttactgctgt tttattcgca
      661 gcatcctccg cattagctgc tccagtcaac actacaacag aagatgaaac ggcacaaatt
      721 ccggctgaag ctgtcatcgg ttactcagat ttagaagggg atttcgatgt tgctgttttg
      781 ccattttcca acagcacaaa taacgggtta ttgtttataa atactactat tgccagcatt
      841 gctgctaaag aagaaggggt atctctcgag aaaagagaag ctgaagccca attgttcacg
      901 tggcccagcc ggccgtctcg gatcggtacc cgcggcggcc gccagcttgg gcccaattgg
      961 ggtggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgggtgt cgaccatcat
     1021 catcatcatc attgagtttg tagccttaga catgactgtt cctcagttca agttgggcac
     1081 ttacgagaag accggtcttg ctagattcta atcaagagga tgtcagaatg ccatttgcct
     1141 gagagatgca ggcttcattt ttgatacttt tttatttgta acctatatag tataggattt
     1201 tttttgtcat tttgtttctt ctcgtacgag cttgctcctg atcagcctat ctcgcagctg
     1261 atgaatatct tgtggtaggg gtttgggaaa atcattcgag tttgatgttt ttcttggtat
     1321 ttcccactcc tcttcagagt acagaagatt aagtgagacc ttcgtttgtg cggatccggc
     1381 gcgccgatcc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     1441 atatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     1501 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     1561 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     1621 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     1681 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1741 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     1801 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     1861 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     1921 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     1981 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     2041 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     2101 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     2161 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     2221 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     2281 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     2341 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     2401 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     2461 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     2521 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     2581 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     2641 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     2701 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     2761 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     2821 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2881 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2941 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3001 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3061 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3121 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3181 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3241 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3301 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3361 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3421 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3481 tggccttttg ctggcctttt gctcacacac catagcttca aaatgtttct actccttttt
     3541 tactcttcca gattttctcg gactccgcgc atcgccgtac cacttcaaaa cacccaagca
     3601 cagcatacta aatttcccct ctttcttcct ctagggtgtc gttaattacc cgtactaaag
     3661 gtttggaaaa gaaaaaagag accgcctcgt ttctttttct tcgtcgaaaa aggcaataaa
     3721 aatttttatc acgtttcttt ttcttgaaaa tttttttttt tgattttttt ctctttcgat
     3781 gacctcccat tgatatttaa gttaataaac ggtcttcaat ttctcaagtt tcagtttcat
     3841 ttttcttgtt ctattacaac tttttttact tcttgctcat tagaaagaaa gcatagcaat
     3901 ctaatctaag ggcggcgttg acaattaatc atcggcatag tatatcggca tagtataata
     3961 cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc
     4021 gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt
     4081 cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt
     4141 ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga
     4201 gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc
     4261 catgaccgag atcggcgagc agccgtgggg gcaggagttc gccctgcgcg acccggccgg
     4321 caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtccgacgcg gcccgacggg
     4381 tccgaggcct cggagatccg tccccctttt cctttgtcga tatcatgtaa ttagttatgt
     4441 cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac
     4501 aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat
     4561 ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac
     4621 tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc aagctgaatt
     4681 gcggccgcag atct
//
