LOCUS       pLyGo-Kp-2      9505 bp    DNA     circular UNA 05-NOV-2020
DEFINITION  .
KEYWORDS    "creator:Thermo Fisher (Invitrogen)" "marker:AmpR, KanR, USER,     
            USER" "organism:Pichia pastoris"
FEATURES             Location/Qualifiers
     promoter        2..937
                     /label="AOX1 promoter"
     sig_peptide     949..1008
                     /label="α-amylase signal sequence"
                     /label="AmySP"
     CDS             complement(1024..1329)
                     /label="ccdB"
     promoter        complement(1330..1432)
                     /label="PccdB"
     terminator      complement(1433..1466)
                     /label="Bba_B1002 Terminator"
     terminator      1550..1796
                     /label="AOX1 terminator"
     CDS             complement(2209..4743)
                     /label="PpHIS4"
     CDS             complement(5157..5972)
                     /label="KanR"
     misc_feature    6351..7107
                     /label="AOX1 3' fragment"
     rep_origin      complement(7576..8164)
                     /label="ori"
     CDS             complement(8335..9195)
                     /label="AmpR"
     promoter        complement(9196..9300)
                     /label="AmpR promoter"
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 AAAGTCGCCA 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 ATTCGAAGGA TCCAAACGAT GGTCGCTTGG
      961 TGGTCTTTGT TTCTGTACGG TCTTCAGGTC GCTGCACCTG CTTTGGCTCA TCGAAGAGCT
     1021 TAATTATATT CCCCAGAACA TCAGGTTAAT GGCGTTTTTG ATGTCATTTT CGCGGTGGCT
     1081 GAGATCAGCC ACTTCTTCCC CGATAACGGA GACCGGCACA CTGGCCATAT CGGTGGTCAT
     1141 CATGCGCCAG CTTTTATCCC CGATATGCAC CACCGGGTAA AGTTCACGGG AGACTTTATC
     1201 TGACAGCAGA CGTGCACTGG CCAGGGGGAT CACCATCCGT CGCCCGGGCG TGTCAATAAT
     1261 ATCACTCTGT ACATCCACAA ACAGACGATA ACGGCTCTCT CTTTTATAGG TGTAAACCTT
     1321 AAACTGCATA GCATACCTCT TTTTGACATA CTTCGGGTAT ACATATCAGT ATATATTCTT
     1381 ATACCGCAAA AATCAGCGCG CAAATACGCA TACTGTTATC TGGCTTTTAG TAGCGAAAAA
     1441 ACCCCGCCGA AGCGGGGTTT TTTGCGGCTC TTCGTAATTC GCCTTAGACA TGACTGTTCC
     1501 TCAGTTCAAG TTGGGCACTT ACGAGAAGAC CGGTCTTGCT AGATTCTAAT CAAGAGGATG
     1561 TCAGAATGCC ATTTGCCTGA GAGATGCAGG CTTCATTTTT GATACTTTTT TATTTGTAAC
     1621 CTATATAGTA TAGGATTTTT TTTGTCATTT TGTTTCTTCT CGTACGAGCT TGCTCCTGAT
     1681 CAGCCTATCT CGCAGCTGAT GAATATCTTG TGGTAGGGGT TTGGGAAAAT CATTCGAGTT
     1741 TGATGTTTTT CTTGGTATTT CCCACTCCTC TTCAGAGTAC AGAAGATTAA GTGAGAAGTT
     1801 CGTTTGTGCA AGCTTATCGA TAAGCTTTAA TGCGGTAGTT TATCACAGTT AAATTGCTAA
     1861 CGCAGTCAGG CACCGTGTAT GAAATCTAAC AATGCGCTCA TCGTCATCCT CGGCACCGTC
     1921 ACCCTGGATG CTGTAGGCAT AGGCTTGGTT ATGCCGGTAC TGCCGGGCCT CTTGCGGGAT
     1981 ATCGTCCATT CCGACAGCAT CGCCAGTCAC TATGGCGTGC TGCTAGCGCT ATATGCGTTG
     2041 ATGCAATTTC TATGCGCACC CGTTCTCGGA GCACTGTCCG ACCGCTTTGG CCGCCGCCCA
     2101 GTCCTGCTCG CTTCGCTACT TGGAGCCACT ATCGACTACG CGATCATGGC GACCACACCC
     2161 GTCCTGTGGA TCTATCGAAT CTAAATGTAA GTTAAAATCT CTAAATAATT AAATAAGTCC
     2221 CAGTTTCTCC ATACGAACCT TAACAGCATT GCGGTGAGCA TCTAGACCTT CAACAGCAGC
     2281 CAGATCCATC ACTGCTTGGC CAATATGTTT CAGTCCCTCA GGAGTTACGT CTTGTGAAGT
     2341 GATGAACTTC TGGAAGGTTG CAGTGTTAAC TCCGCTGTAT TGACGGGCAT ATCCGTACGT
     2401 TGGCAAAGTG TGGTTGGTAC CGGAGGAGTA ATCTCCACAA CTCTCTGGAG AGTAGGCACC
     2461 AACAAACACA GATCCAGCGT GTTGTACTTG ATCAACATAA GAAGAAGCAT TCTCGATTTG
     2521 CAGGATCAAG TGTTCAGGAG CGTACTGATT GGACATTTCC AAAGCCTGCT CGTAGGTTGC
     2581 AACCGATAGG GTTGTAGAGT GTGCAATACA CTTGCGTACA ATTTCAACCC TTGGCAACTG
     2641 CACAGCTTGG TTGTGAACAG CATCTTCAAT TCTGGCAAGC TCCTTGTCTG TCATATCGAC
     2701 AGCCAACAGA ATCACCTGGG AATCAATACC ATGTTCAGCT TGAGACAGAA GGTCTGAGGC
     2761 AACGAAATCT GGATCAGCGT ATTTATCAGC AATAACTAGA ACTTCAGAAG GCCCAGCAGG
     2821 CATGTCAATA CTACACAGGG CTGATGTGTC ATTTTGAACC ATCATCTTGG CAGCAGTAAC
     2881 GAACTGGTTT CCTGGACCAA ATATTTTGTC ACACTTAGGA ACAGTTTCTG TTCCGTAAGC
     2941 CATAGCAGCT ACTGCCTGGG CGCCTCCTGC TAGCACGATA CACTTAGCAC CAACCTTGTG
     3001 GGCAACGTAG ATGACTTCTG GGGTAAGGGT ACCATCCTTC TTAGGTGGAG ATGCAAAAAC
     3061 AATTTCTTTG CAACCAGCAA CTTTGGCAGG AACACCCAGC ATCAGGGAAG TGGAAGGCAG
     3121 AATTGCGGTT CCACCAGGAA TATAGAGGCC AACTTTCTCA ATAGGTCTTG CAAAACGAGA
     3181 GCAGACTACA CCAGGGCAAG TCTCAACTTG CAACGTCTCC GTTAGTTGAG CTTCATGGAA
     3241 TTTCCTGACG TTATCTATAG AGAGATCAAT GGCTCTCTTA ACGTTATCTG GCAATTGCAT
     3301 AAGTTCCTCT GGGAAAGGAG CTTCTAACAC AGGTGTCTTC AAAGCGACTC CATCAAACTT
     3361 GGCAGTTAGT TCTAAAAGGG CTTTGTCACC ATTTTGACGA ACATTGTCGA CAATTGGTTT
     3421 GACTAATTCC ATAATCTGTT CCGTTTTCTG GATAGGACGA CGAAGGGCAT CTTCAATTTC
     3481 TTGTGAGGAG GCCTTAGAAA CGTCAATTTT GCACAATTCA ATACGACCTT CAGAAGGGAC
     3541 TTCTTTAGGT TTGGATTCTT CTTTAGGTTG TTCCTTGGTG TATCCTGGCT TGGCATCTCC
     3601 TTTCCTTCTA GTGACCTTTA GGGACTTCAT ATCCAGGTTT CTCTCCACCT CGTCCAACGT
     3661 CACACCGTAC TTGGCACATC TAACTAATGC AAAATAAAAT AAGTCAGCAC ATTCCCAGGC
     3721 TATATCTTCC TTGGATTTAG CTTCTGCAAG TTCATCAGCT TCCTCCCTAA TTTTAGCGTT
     3781 CAACAAAACT TCGTCGTCAA ATAACCGTTT GGTATAAGAA CCTTCTGGAG CATTGCTCTT
     3841 ACGATCCCAC AAGGTGGCTT CCATGGCTCT AAGACCCTTT GATTGGCCAA AACAGGAAGT
     3901 GCGTTCCAAG TGACAGAAAC CAACACCTGT TTGTTCAACC ACAAATTTCA AGCAGTCTCC
     3961 ATCACAATCC AATTCGATAC CCAGCAACTT TTGAGTTGCT CCAGATGTAG CACCTTTATA
     4021 CCACAAACCG TGACGACGAG ATTGGTAGAC TCCAGTTTGT GTCCTTATAG CCTCCGGAAT
     4081 AGACTTTTTG GACGAGTACA CCAGGCCCAA CGAGTAATTA GAAGAGTCAG CCACCAAAGT
     4141 AGTGAATAGA CCATCGGGGC GGTCAGTAGT CAAAGACGCC AACAAAATTT CACTGACAGG
     4201 GAACTTTTTG ACATCTTCAG AAAGTTCGTA TTCAGTAGTC AATTGCCGAG CATCAATAAT
     4261 GGGGATTATA CCAGAAGCAA CAGTGGAAGT CACATCTACC AACTTTGCGG TCTCAGAAAA
     4321 AGCATAAACA GTTCTACTAC CGCCATTAGT GAAACTTTTC AAATCGCCCA GTGGAGAAGA
     4381 AAAAGGCACA GCGATACTAG CATTAGCGGG CAAGGATGCA ACTTTATCAA CCAGGGTCCT
     4441 ATAGATAACC CTAGCGCCTG GGATCATCCT TTGGACAACT CTTTCTGCCA AATCTAGGTC
     4501 CAAAATCACT TCATTGATAC CATTATTGTA CAACTTGAGC AAGTTGTCGA TCAGCTCCTC
     4561 AAATTGGTCC TCTGTAACGG ATGACTCAAC TTGCACATTA ACTTGAAGCT CAGTCGATTG
     4621 AGTGAACTTG ATCAGGTTGT GCAGCTGGTC AGCAGCATAG GGAAACACGG CTTTTCCTAC
     4681 CAAACTCAAG GAATTATCAA ACTCTGCAAC ACTTGCGTAT GCAGGTAGCA AGGGAAATGT
     4741 CATACTTGAA GTCGGACAGT GAGTGTAGTC TTGAGAAATT CTGAAGCCGT ATTTTTATTA
     4801 TCAGTGAGTC AGTCATCAGG AGATCCTCTA CGCCGGACGC ATCGTGGCCG ACCTGCAGGG
     4861 GGGGGGGGGG CGCTGAGGTC TGCCTCGTGA AGAAGGTGTT GCTGACTCAT ACCAGGCCTG
     4921 AATCGCCCCA TCATCCAGCC AGAAAGTGAG GGAGCCACGG TTGATGAGAG CTTTGTTGTA
     4981 GGTGGACCAG TTGGTGATTT TGAACTTTTG CTTTGCCACG GAACGGTCTG CGTTGTCGGG
     5041 AAGATGCGTG ATCTGATCCT TCAACTCAGC AAAAGTTCGA TTTATTCAAC AAAGCCGCCG
     5101 TCCCGTCAAG TCAGCGTAAT GCTCTGCCAG TGTTACAACC AATTAACCAA TTCTGATTAG
     5161 AAAAACTCAT CGAGCATCAA ATGAAACTGC AATTTATTCA TATCAGGATT ATCAATACCA
     5221 TATTTTTGAA AAAGCCGTTT CTGTAATGAA GGAGAAAACT CACCGAGGCA GTTCCATAGG
     5281 ATGGCAAGAT CCTGGTATCG GTCTGCGATT CCGACTCGTC CAACATCAAT ACAACCTATT
     5341 AATTTCCCCT CGTCAAAAAT AAGGTTATCA AGTGAGAAAT CACCATGAGT GACGACTGAA
     5401 TCCGGTGAGA ATGGCAAAAG CTTATGCATT TCTTTCCAGA CTTGTTCAAC AGGCCAGCCA
     5461 TTACGCTCGT CATCAAAATC ACTCGCATCA ACCAAACCGT TATTCATTCG TGATTGCGCC
     5521 TGAGCGAGAC GAAATACGCG ATCGCTGTTA AAAGGACAAT TACAAACAGG AATCGAATGC
     5581 AACCGGCGCA GGAACACTGC CAGCGCATCA ACAATATTTT CACCTGAATC AGGATATTCT
     5641 TCTAATACCT GGAATGCTGT TTTCCCGGGG ATCGCAGTGG TGAGTAACCA TGCATCATCA
     5701 GGAGTACGGA TAAAATGCTT GATGGTCGGA AGAGGCATAA ATTCCGTCAG CCAGTTTAGT
     5761 CTGACCATCT CATCTGTAAC ATCATTGGCA ACGCTACCTT TGCCATGTTT CAGAAACAAC
     5821 TCTGGCGCAT CGGGCTTCCC ATACAATCGA TAGATTGTCG CACCTGATTG CCCGACATTA
     5881 TCGCGAGCCC ATTTATACCC ATATAAATCA GCATCCATGT TGGAATTTAA TCGCGGCCTC
     5941 GAGCAAGACG TTTCCCGTTG AATATGGCTC ATAACACCCC TTGTATTACT GTTTATGTAA
     6001 GCAGACAGTT TTATTGTTCA TGATGATATA TTTTTATCTT GTGCAATGTA ACATCAGAGA
     6061 TTTTGAGACA CAACGTGGCT TTCCCCCCCC CCCCTGCAGG TCGGCATCAC CGGCGCCACA
     6121 GGTGCGGTTG CTGGCGCCTA TATCGCCGAC ATCACCGATG GGGAAGATCG GGCTCGCCAC
     6181 TTCGGGCTCA TGAGCGCTTG TTTCGGCGTG GGTATGGTGG CAGGCCCCGT GGCCGGGGGA
     6241 CTGTTGGGCG CCATCTCCTT GCATGCACCA TTCCTTGCGG CGGCGGTGCT CAACGGCCTC
     6301 AACCTACTAC TGGGCTGCTT CCTAATGCAG GAGTCGCATA AGGGAGAGCG TCGAGTATCT
     6361 ATGATTGGAA GTATGGGAAT GGTGATACCC GCATTCTTCA GTGTCTTGAG GTCTCCTATC
     6421 AGATTATGCC CAACTAAAGC AACCGGAGGA GGAGATTTCA TGGTAAATTT CTCTGACTTT
     6481 TGGTCATCAG TAGACTCGAA CTGTGAGACT ATCTCGGTTA TGACAGCAGA AATGTCCTTC
     6541 TTGGAGACAG TAAATGAAGT CCCACCAATA AAGAAATCCT TGTTATCAGG AACAAACTTC
     6601 TTGTTTCGAA CTTTTTCGGT GCCTTGAACT ATAAAATGTA GAGTGGATAT GTCGGGTAGG
     6661 AATGGAGCGG GCAAATGCTT ACCTTCTGGA CCTTCAAGAG GTATGTAGGG TTTGTAGATA
     6721 CTGATGCCAA CTTCAGTGAC AACGTTGCTA TTTCGTTCAA ACCATTCCGA ATCCAGAGAA
     6781 ATCAAAGTTG TTTGTCTACT ATTGATCCAA GCCAGTGCGG TCTTGAAACT GACAATAGTG
     6841 TGCTCGTGTT TTGAGGTCAT CTTTGTATGA ATAAATCTAG TCTTTGATCT AAATAATCTT
     6901 GACGAGCCAA GGCGATAAAT ACCCAAATCT AAAACTCTTT TAAAACGTTA AAAGGACAAG
     6961 TATGTCTGCC TGTATTAAAC CCCAAATCAG CTCGTAGTCT GATCCTCATC AACTTGAGGG
     7021 GCACTATCTT GTTTTAGAGA AATTTGCGGA GATGCGATAT CGAGAAAAAG GTACGCTGAT
     7081 TTTAAACGTG AAATTTATCT CAAGATCTCT GCCTCGCGCG TTTCGGTGAT GACGGTGAAA
     7141 ACCTCTGACA CATGCAGCTC CCGGAGACGG TCACAGCTTG TCTGTAAGCG GATGCCGGGA
     7201 GCAGACAAGC CCGTCAGGGC GCGTCAGCGG GTGTTGGCGG GTGTCGGGGC GCAGCCATGA
     7261 CCCAGTCACG TAGCGATAGC GGAGTGTATA CTGGCTTAAC TATGCGGCAT CAGAGCAGAT
     7321 TGTACTGAGA GTGCACCATA TGCGGTGTGA AATACCGCAC AGATGCGTAA GGAGAAAATA
     7381 CCGCATCAGG CGCTCAACCG CTTCCTCGCT CACTGACTCG CTGCGCTCGG TCGTTCGGCT
     7441 GCGGCGAGCG GTATCAGCTC ACTCAAAGGC GGTAATACGG TTATCCACAG AATCAGGGGA
     7501 TAACGCAGGA AAGAACATGT GAGCAAAAGG CCAGCAAAAG GCCAGGAACC GTAAAAAGGC
     7561 CGCGTTGCTG GCGTTTTTCC ATAGGCTCCG CCCCCCTGAC GAGCATCACA AAAATCGACG
     7621 CTCAAGTCAG AGGTGGCGAA ACCCGACAGG ACTATAAAGA TACCAGGCGT TTCCCCCTGG
     7681 AAGCTCCCTC GTGCGCTCTC CTGTTCCGAC CCTGCCGCTT ACCGGATACC TGTCCGCCTT
     7741 TCTCCCTTCG GGAAGCGTGG CGCTTTCTCA ATGCTCACGC TGTAGGTATC TCAGTTCGGT
     7801 GTAGGTCGTT CGCTCCAAGC TGGGCTGTGT GCACGAACCC CCCGTTCAGC CCGACCGCTG
     7861 CGCCTTATCC GGTAACTATC GTCTTGAGTC CAACCCGGTA AGACACGACT TATCGCCACT
     7921 GGCAGCAGCC ACTGGTAACA GGATTAGCAG AGCGAGGTAT GTAGGCGGTG CTACAGAGTT
     7981 CTTGAAGTGG TGGCCTAACT ACGGCTACAC TAGAAGGACA GTATTTGGTA TCTGCGCTCT
     8041 GCTGAAGCCA GTTACCTTCG GAAAAAGAGT TGGTAGCTCT TGATCCGGCA AACAAACCAC
     8101 CGCTGGTAGC GGTGGTTTTT TTGTTTGCAA GCAGCAGATT ACGCGCAGAA AAAAAGGATC
     8161 TCAAGAAGAT CCTTTGATCT TTTCTACGGG GTCTGACGCT CAGTGGAACG AAAACTCACG
     8221 TTAAGGGATT TTGGTCATGA GATTATCAAA AAGGATCTTC ACCTAGATCC TTTTAAATTA
     8281 AAAATGAAGT TTTAAATCAA TCTAAAGTAT ATATGAGTAA ACTTGGTCTG ACAGTTACCA
     8341 ATGCTTAATC AGTGAGGCAC CTATCTCAGC GATCTGTCTA TTTCGTTCAT CCATAGTTGC
     8401 CTGACTCCCC GTCGTGTAGA TAACTACGAT ACGGGAGGGC TTACCATCTG GCCCCAGTGC
     8461 TGCAATGATA CCGCGAGACC CACGCTCACC GGCTCCAGAT TTATCAGCAA TAAACCAGCC
     8521 AGCCGGAAGG GCCGAGCGCA GAAGTGGTCC TGCAACTTTA TCCGCCTCCA TCCAGTCTAT
     8581 TAATTGTTGC CGGGAAGCTA GAGTAAGTAG TTCGCCAGTT AATAGTTTGC GCAACGTTGT
     8641 TGCCATTGCT GCAGGCATCG TGGTGTCACG CTCGTCGTTT GGTATGGCTT CATTCAGCTC
     8701 CGGTTCCCAA CGATCAAGGC GAGTTACATG ATCCCCCATG TTGTGCAAAA AAGCGGTTAG
     8761 CTCCTTCGGT CCTCCGATCG TTGTCAGAAG TAAGTTGGCC GCAGTGTTAT CACTCATGGT
     8821 TATGGCAGCA CTGCATAATT CTCTTACTGT CATGCCATCC GTAAGATGCT TTTCTGTGAC
     8881 TGGTGAGTAC TCAACCAAGT CATTCTGAGA ATAGTGTATG CGGCGACCGA GTTGCTCTTG
     8941 CCCGGCGTCA ACACGGGATA ATACCGCGCC ACATAGCAGA ACTTTAAAAG TGCTCATCAT
     9001 TGGAAAACGT TCTTCGGGGC GAAAACTCTC AAGGATCTTA CCGCTGTTGA GATCCAGTTC
     9061 GATGTAACCC ACTCGTGCAC CCAACTGATC TTCAGCATCT TTTACTTTCA CCAGCGTTTC
     9121 TGGGTGAGCA AAAACAGGAA GGCAAAATGC CGCAAAAAAG GGAATAAGGG CGACACGGAA
     9181 ATGTTGAATA CTCATACTCT TCCTTTTTCA ATATTATTGA AGCATTTATC AGGGTTATTG
     9241 TCTCATGAGC GGATACATAT TTGAATGTAT TTAGAAAAAT AAACAAATAG GGGTTCCGCG
     9301 CACATTTCCC CGAAAAGTGC CACCTGACGT CTAAGAAACC ATTATTATCA TGACATTAAC
     9361 CTATAAAAAT AGGCGTATCA CGAGGCCCTT TCGTCTTCAA GAATTAATTC TCATGTTTGA
     9421 CAGCTTATCA TCGATAAGCT GACTCATGTT GGTATTGTGA AATAGACGCA GATCGGGAAC
     9481 ACTGAAAAAT AACAGTTATT ATTCG
//
