LOCUS       40924_35743        9139 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pPYPX99, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9139)
  AUTHORS   Yao Q, Peng R, Xiong A.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-NOV-2002) Shanghai Academy of Agricultural Sciences, 
            Agro-Biotechnology Research Center, 2901 Beidi Rd, Shanghai 201106, 
            China
REFERENCE   2  (bases 1 to 9139)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9139)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (12-NOV-2002) Shanghai Academy of Agricultural Sciences, 
            Agro-Biotechnology Research Center, 2901 Beidi Rd, Shanghai 201106, 
            China"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     NCBI staff are still waiting for submitters to provide appropriate 
            coding region information.
FEATURES             Location/Qualifiers
     source          1..9139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..936
                     /label=modified AOX1 promoter
                     /note="modified AOX1 promoter"
     misc_feature    937..1212
                     /label=similar to modified alpha factor secretion signal
                     /note="similar to modified alpha factor secretion signal"
     terminator      1343..1589
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     misc_feature    complement(1994..4527)
                     /label=similar to phosphoribosyl-ATP pyrophosphohydrolase
                     /note="similar to phosphoribosyl-ATP pyrophosphohydrolase"
     CDS             4798..5610
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_feature    5985..6741
                     /label=AOX1 3' fragment
                     /note="region downstream of Pichia pastoris AOX1 gene"
     misc_feature    6884..7024
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(7210..7798)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7972..8829)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8830..8934)
                     /label=AmpR promoter
     promoter        9139
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
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 ttggcgctca ttccaattcc ttctattagg ctactaacac
      241 catgacttta ttagcctgtc tatcctggcc cccctggcga ggttcatgtt tgtttatttc
      301 cgaatgcaac aagctccgca ttacacccga acatcactcc agatgagggc tttctgagtg
      361 tggggtcaaa tagtttcatg ttccccaaat ggcccaaaac tgacagttta aacgctgtct
      421 tggaacctaa tatgacaaaa gcgtgatctc atccaagatg aactaagttt ggttcgttga
      481 aatgctaacg gccagttggt caaaaagaaa cttccaaaag tcgccatacc gtttgtcttg
      541 tttggtattg attgacgaat gctcaaaaat aatctcatta atgcttagcg cagtctctct
      601 atcgcttctg aaccccggtg cacctgtgcc gaaacgcaaa tggggaaaca cccgcttttt
      661 ggatgattat gcattgtctc cacattgtat gcttccaaga ttctggtggg aatactgctg
      721 atagcctaac gttcatgatc aaaatttaac tgttctaacc cctacttgac agcaatatat
      781 aaacagaagg aagctgccct gtcttaaacc ttttttttta tcatcattat tagcttactt
      841 tcataattgc gactggttcc aattgacaag cttttgattt taacgacttt taacgacaac
      901 ttgagaagat caaaaaacaa ctaattattc gaaaccatgg ctattccacc atctatcttc
      961 atcgctgtct tgttcgctgc ttcttctgcc ttggctgctc ctgtcaacac tactaccgag
     1021 gatgaaactg ctcaaatccc tgctgaggct gtcatcggtt actctgacct ggagggtgac
     1081 ttcgacgtcg ctgtcttgcc attctctaac tccaccaaca acggtttgtt ggaggaggct
     1141 gaagctgaag ctgaacctaa attcatcaac actactatcg cttctatcgc tgctaaggag
     1201 gagggtgttt ccctcgagaa aagagaggct gaggcttacg tagagttgag ctcgggcggc
     1261 cgcgaattaa ttcgccttag acatgactgt tcctcagttc aagttgggca cttacgagaa
     1321 gaccggtctt gctagattct aatcaagagg atgtcagaat gccatttgcc tgagagatgc
     1381 aggcttcatt tttgatactt ttttatttgt aacctatata gtataggatt ttttttgtca
     1441 ttttgtttct tctcgtacga gcttgctcct gatcagccta tctcgcagct gatgaatatc
     1501 ttgtggtagg ggtttgggaa aatcattcga gtttgatgtt tttcttggta tttcccactc
     1561 ctcttcagag tacagaagat taagtgagaa gttcgtttgt gcaagctagc tttaatgcgg
     1621 tagtttatca cagttaaatt gctaacgcag tcaggcaccg tgtatgaaat ctaacaatgc
     1681 gctcatcgtc atcctcggca ccgtcaccct ggatgctgta ggcataggct tggttatgcc
     1741 ggtactgccg ggcctcttgc gggatatcgt ccattccgac agcatcgcca gtcactatgg
     1801 cgtgctgcta gcgctatatg cgttgatgca atttctatgc gcacccgttc tcggagcact
     1861 gtccgaccgc tttggccgcc gcccagtcct gctcgcttcg ctacttggag ccactatcga
     1921 ctacgcgatc atggcgacca cacccgtcct gtggatctat cgaatctaaa tgtaagttaa
     1981 aatctctaaa taattaaata agtcccagtt tctccatacg aaccttaaca gcattgcggt
     2041 gagcatctag accttcaaca gcagccagat ccatcactgc ttggccaata tgtttcagtc
     2101 cctcaggagt tacgtcttgt gaagtgatga acttctggaa ggttgcagtg ttaactccgc
     2161 tgtattgacg ggcatatccg tacgttggca aagtgtggtt ggtaccggag gagtaatctc
     2221 cacaactctc tggagagtag gcaccaacaa acacagatcc agcgtgttgt acttgatcaa
     2281 cataagaaga agcattctcg atttgcagga tcaagtgttc aggagcgtac tgattggaca
     2341 tttccaaagc ctgctcgtag gttgcaaccg atagggttgt agagtgtgca atacacttgc
     2401 gtacaatttc aacccttggc aactgcacag cttggttgtg aacagcatct tcaattctgg
     2461 caagctcctt gtctgtcata tcgacagcca acagaatcac ctgggaatca ataccatgtt
     2521 cagcttgagc agaaggtctg aggcaacgaa atctggatca gcgtatttat cagcaataac
     2581 tagaacttca gaaggcccag caggcatgtc aatactacac agggctgatg tgtcattttg
     2641 aaccatcatc ttggcagcag taacgaactg gtttcctgga ccaaatattt tgtcacactt
     2701 aggaacagtt tctgttccgt aagccatagc agctactgcc tgggcgcctc ctgctagcac
     2761 gatacactta gcaccaacct tgtgggcaac gtagatgact tctggggtaa gggtaccatc
     2821 cttcttaggt ggagatgcaa aaacaatttc tttgcaacca gcaactttgg caggaacacc
     2881 cagcatcagg gaagtggaag gcagaattgc ggttccacca ggaatataga ggccaacttt
     2941 ctcaataggt cttgcaaaac gagagcagac tacaccaggg caagtctcaa cttgcaacgt
     3001 ctccgttagt tgagcttcat ggaatttcct gacgttatct atagagagat caatggctct
     3061 cttaacgtta tctggcaatt gcataagttc ctctgggaaa ggagcttcta acacaggtgt
     3121 cttcaaagcg actccatcaa acttggcagt tagttctaaa agggctttgt caccattttg
     3181 acgaacattg tcgacaattg gtttgactaa ttccataatc tgttccgttt tctggatagg
     3241 acgacgaagg gcatcttcaa tttcttgtga ggaggcctta gaaacgtcaa ttttgcacaa
     3301 ttcaatacga ccttcagaag ggacttcttt aggtttggat tcttctttag gttgttcctt
     3361 ggtgtatcct ggcttggcat ctcctttcct tctagtgacc tttagggact tcatatccag
     3421 gtttctctcc acctcgtcca acgtcacacc gtacttggca catctaacta atgcaaaata
     3481 aaataagtca gcacattccc aggctatatc ttccttggat ttagcttctg caagttcatc
     3541 agcttcctcc ctaattttag cgttcaaaca aaacttcgtc gtcaaataac cgtttggtat
     3601 aagaaccttc tggagcattg ctcttacgat cccacaaggt gcttccatgg ctctaagacc
     3661 ctttgattgg ccaaaacagg aagtgcgttc caagtgacag aaaccaacac ctgtttgttc
     3721 aaccacaaat ttcaagcagt ctccatcaca atccaattcg atacccagca acttttgagt
     3781 tcgtccagat gtagcacctt tataccacaa accgtgacga cgagattggt agactccagt
     3841 ttgtgtcctt atagcctccg gaatagactt tttggacgag tacaccaggc ccaacgagta
     3901 attagaagag tcagccacca aagtagtgaa tagaccatcg gggcggtcag tagtcaaaga
     3961 cgccaacaaa atttcactga cagggaactt tttgacatct tcagaaagtt cgtattcagt
     4021 agtcaattgc cgagcatcaa taatggggat tataccagaa gcaacagtgg aagtcacatc
     4081 taccaacttt gcggtctcag aaaaagcata aacagttcta ctaccgccat tagtgaaact
     4141 tttcaaatcg cccagtggag aagaaaaagg cacagcgata ctagcattag cgggcaagga
     4201 tgcaacttta tcaaccaggg tcctatagat aaccctagcg cctgggatca tcctttggac
     4261 aactctttct gccaaatcta ggtccaaaat cacttcattg ataccattat tgtacaactt
     4321 gagcaagttg tcgatcagct cctcaaattg gtcctctgta acggatgact caacttgcac
     4381 attaacttga agctcagtcg attgagtgaa cttgatcagg ttgtgcagct ggtcagcagc
     4441 atagggaaac acggcttttc ctaccaaact caaggaatta tcaaactctg caacacttgc
     4501 gtatgcaggt agcaagggaa atgtcatact tgaagtcgga cagtgagtgt agtcttgaga
     4561 aattctgaag ccgtattttt attatcagtg agtcagtcat caggagatcc tctacgccgg
     4621 acgcatcgtg gccgatggtg cttgggtgaa caggtttatt cctggcatcc actaaatata
     4681 gcatgcgaaa gccacgttgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     4741 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     4801 agccatattc aacgggaaac gtcttgttcg aggccgcgat taaattccaa catggatgct
     4861 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     4921 cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     4981 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     5041 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     5101 ccagggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     5161 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     5221 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     5281 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     5341 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     5401 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     5461 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     5521 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     5581 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     5641 tggcagagca ttacgctgac ttgacgggac ggcggctttg ttgaataaat cgaacttttg
     5701 ctgagttgaa ggatcagatc acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc
     5761 aaaagttcaa aatcaccaac tggtccacct acaacaaagc tctcatcaac cgtggctccc
     5821 tcactttctg gctggatgat ggggcgattc aggcctggta tgagtcagca acaccttctt
     5881 cacgaggcag acctcagcgc cgcggattcc ttgcggcggc ggtgctaacg gcctcaacct
     5941 acgcgtggct gcttcctaat gcaggagtcg cataagggag agcgtcgagt atctatgatt
     6001 ggaagtatgg gaatggtgat acccgcattc ttcagtgtct tgaggtctcc tatcagatta
     6061 tgcccaacta aagcaaccgg aggaggagat ttcatggtaa atttctctga cttttggtca
     6121 tcagtagact cgaactgtga gactatctcg gttatgacag cagaaatgtc cttcttggag
     6181 acagtaaatg aagtcccacc aataaagaaa tccttgttat caggaacaaa cttcttgttt
     6241 cgaacttttt cggtgccttg aactataaaa tgtagagtgg atatgtcggg taggaatgga
     6301 gcgggcaaat gcttaccttc tggaccttca agaggtatgt agggtttgta gatactgatg
     6361 ccaacttcag tgacaacgtt gctatttcgt tcaaaccatt ccgaatccag agaaatcaaa
     6421 gttgtttgtc tactattgat ccaagccagt gcggtcttga aactgacaat agtgtgctcg
     6481 tgttttgagg tcatctttgt atgaataaat ctagtctttg atctaaataa tcttgacgag
     6541 ccaaggcgat aaatacccaa atctaaaact cttttaaaac gttaaaagga caagtatgtc
     6601 tgcctgtatt aaaccccaaa tcagctcgta gtctgatcct catcaacttg aggggcacta
     6661 tcttgtttta gagaaatttg cggagatgcg atatcgagaa aaaggtacgc tgattttaaa
     6721 cgtgaaattt atctcaagat ctctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     6781 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     6841 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     6901 cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact
     6961 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     7021 caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7081 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7141 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7201 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7261 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7321 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7381 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     7441 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7501 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7561 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7621 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     7681 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7741 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7801 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7861 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7921 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7981 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8041 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8101 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8161 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8221 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8281 tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8341 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8401 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8461 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8521 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8581 gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8641 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8701 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8761 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8821 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8881 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8941 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     9001 aaataggcgt atcacgaggc cctttcgtct tcaagaatta attctcatgt ttgacagctt
     9061 atcatcgata agctgactca tgttggtatt gtgaaataga cgcagatcgg gaacactgaa
     9121 aaataacagt tattattcg
//
