LOCUS       40924_26531        4558 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Extracellular expression vector pKanB alpha, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4558)
  AUTHORS   Lin-Cereghino J, Hashimoto MD, Moy A, Castelo J, Orazem CC, Kuo P, 
            Xiong S, Gandhi V, Hatae CT, Chan A, Lin-Cereghino GP.
  TITLE     Direct selection of Pichia pastoris expression strains using new 
            G418 resistance vectors
  JOURNAL   Yeast 25 (4), 293-299 (2008)
  PUBMED    18327886
REFERENCE   2  (bases 1 to 4558)
  AUTHORS   Lin-Cereghino G, Lin-Cereghino J.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-NOV-2007) Biological Sciences, University of the 
            Pacific, 3601 Pacific Ave, Stockton, CA 95211, USA
REFERENCE   3  (bases 1 to 4558)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4558)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Yeast"; 
            date: "2008"; volume: "25"; issue: "4"; pages: "293-299"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (17-NOV-2007) Biological Sciences, University of the Pacific, 3601 
            Pacific Ave, Stockton, CA 95211, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4558
                     /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             1279..1308
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     CDS             1324..1341
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      1421..1667
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             2226..3038
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             3060..3089
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     CDS             3105..3122
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      3201..3447
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     terminator      3568..3815
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(3890..4478)
                     /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 tgaagctgca ggaattcacg tggcccagcc ggccgtctcg gatcggtacc tcgagccgcg
     1261 gcggccgcca gctttctaga acaaaaactc atctcagaag aggatctgaa tagcgccgtc
     1321 gaccatcatc atcatcatca ttgagtttgt agccttagac atgactgttc ctcagttcaa
     1381 gttgggcact tacgagaaga ccggtcttgc tagattctaa tcaagaggat gtcagaatgc
     1441 catttgcctg agagatgcag gcttcatttt tgatactttt ttatttgtaa cctatatagt
     1501 ataggatttt ttttgtcatt ttgtttcttc tcgtacgagc ttgctcctga tcagcctatc
     1561 tcgcagctga tgaatatctt gtggtagggg tttgggaaaa tcattcgagt ttgatgtttt
     1621 tcttggtatt tcccactcct cttcagagta cagaagatta agtgagacct tcgtttgtgc
     1681 ggatccttga agctatggtg tgtgggaggg ctaactcgag gtactgattg gacgaggaca
     1741 caggcctggt gtcctcgtcc aatcaggtag ccatctctga aatatctggc tccgttgcaa
     1801 ctccgaacga cctgctggca acgtaaaatt ctccggggta aaacttaaat gtggagtaat
     1861 ggaaccagaa acgtctcttc ccttctctct ccttccaccg cccgttaccg tccctaggaa
     1921 attttactct gctggagagc ttcttctacg gcccccttgc agcaatgctc ttcccagcat
     1981 tacgttgcgg gtaaaacgga ggtcgtgtac ccgacctagc agcccaggga tggaaaagtc
     2041 ccggccgtcg ctggcaataa tagcgggcgg acgcatgtca tgagattatt ggaaaccacc
     2101 agaatcgaat ataaaaggcg aacacctttc ccaattttgg tttctcctga cccaaagact
     2161 ttaaatttaa tttatttgtc cctatttcaa tcaattgaac aactatttcg aaacgaggaa
     2221 ttcttatgag ccatattcaa cgggaaacgt cttgctcgag gccgcgatta aattccaaca
     2281 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     2341 caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     2401 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     2461 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     2521 ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     2581 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     2641 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     2701 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     2761 ggaaagaaat gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt
     2821 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     2881 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     2941 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     3001 ataaattgca gtttcatttg atgctcgatg agtttttcta agcggccgcc agctttctag
     3061 aacaaaaact catctcagaa gaggatctga atagcgccgt cgaccatcat catcatcatc
     3121 attgagtttt agccttagac atgactgttc ctcagttcaa gttgggcact tacgagaaga
     3181 ccggtcttgc tagattctaa tcaagaggat gtcagaatgc catttgcctg agagatgcag
     3241 gcttcatttt tgatactttt ttatttgtaa cctatatagt ataggatttt ttttgtcatt
     3301 ttgtttcttc tcgtacgagc ttgctcctga tcagcctatc tcgcagctga tgaatatctt
     3361 gtggtagggg tttgggaaaa tcattcgagt ttgatgtttt tcttggtatt tcccactcct
     3421 cttcagagta cagaagatta agtgagacct tcgtttgtgc ggatccttga agctatggtg
     3481 tgtgtgggag ggctaactcg aggtactgat tggacgagga cacaggcctg gtcctcggag
     3541 atccgtcccc cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac
     3601 gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     3661 tccctattta tttttttata gttatgttag tattaagaac gttatttata tttcaaattt
     3721 ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg
     3781 agaaggtttt gggacgctcg aaggctttaa tttgcaagct ggagaccaac atgtgagcaa
     3841 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3901 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3961 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4021 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4081 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4141 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4201 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4261 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4321 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4381 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4441 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4501 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagatc
//
