LOCUS       sequence-247138-        4412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast plasmid complementing ALD1 from  Komagataella phaffii.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4412)
  AUTHORS   Liu Y, Bai C, Liu Q, Xu Q, Qian Z, Peng Q, Yu J, Xu M, Zhou X, Zhang
            Y, Cai M
  TITLE     Engineered ethanol-driven biosynthetic system for improving 
            production of acetyl-CoA derived drugs in Crabtree-negative yeast.
  JOURNAL   Metab Eng. 2019 Jul;54:275-284. doi: 10.1016/j.ymben.2019.05.001. 
            Epub 2019 May 9.
  PUBMED    31077813
REFERENCE   2  (bases 1 to 4412)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Metab 
            Eng."; date: "2019-07"; volume: "54:275-284. doi"; pages: " 
            10.1016/j.ymben.2019.05.001. Epub 2019 May 9"
COMMENT     Sequence Label: pZ_P-GAP-ALD1
FEATURES             Location/Qualifiers
     source          1..4412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        61..537
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             2143..2172
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2188..2205
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2285..2531
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2546..2959
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2966..3013
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3032..3406
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      3472..3719
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3488..3506)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3794..4382)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3874..3893)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgcatga gatcagatct
       61 tttttgtaga aatgtcttgg tgtcctcgtc caatcaggta gccatctctg aaatatctgg
      121 ctccgttgca actccgaacg acctgctggc aacgtaaaat tctccggggt aaaacttaaa
      181 tgtggagtaa tggaaccaga aacgtctctt cccttctctc tccttccacc gcccgttacc
      241 gtccctagga aattttactc tgctggagag cttcttctac ggcccccttg cagcaatgct
      301 cttcccagca ttacgttgcg ggtaaaacgg aggtcgtgta cccgacctag cagcccaggg
      361 atggaaaagt cccggccgtc gctggcaata atagcgggcg gacgcatgtc atgagattat
      421 tggaaaccac cagaatcgaa tataaaaggc gaacaccttt cccaattttg gtttctcctg
      481 acccaaagac tttaaattta atttatttgt ccctatttca atcaattgaa caactatatg
      541 cttagaactt ctccagctac taagaaagct ctcaagtcgc agattaacgc cttcaacgtt
      601 gctgccttga gattctactc ctcattgcct ttgcaggttc caattacctt gccaaacggt
      661 aagacctaca atcagccaac aggtttgttt atcaacaatg agttcgttcc ttctaagcaa
      721 ggtaagacct ttgctgtttt aaacccttcc actgaggagg agattactca cgtctacgag
      781 tccagagagg acgacgttga gttagccgtt gcagccgctc aaaaggcttt cgactcaacc
      841 tggtccaccc aggaccctgc tgagagaggt aaggtcttga acaagttggc tgacctgatc
      901 gaggagcact ctgagaccct tgccgccatc gagtccttgg acaacggtaa ggccatttcc
      961 tccgctagag gtgatgttgg tctggttgtc gcctacttga agtcctgtgc cggttgggcc
     1021 gacaaggttt tcggtagagt tgttgaaacc ggaagctccc acttcaacta cgttagaaga
     1081 gagccattgg gtgtttgtgg tcagattatc ccatggaact ttcctcttct gatgtggtcc
     1141 tggaaagttg gtccagcttt ggccactggt aacactgttg tcctgaagac agccgagtct
     1201 actcctctgt ccgccctgta cgtttcccaa ttggtcaagg aggccggtat cccagctggt
     1261 gtccacaaca ttgtgtccgg tttcggtaag attactggtg aagctattgc tactcatcct
     1321 aagatcaaga aggttgcctt cactggttct accgccactg gtcgtcacat catgaaggct
     1381 gctgccgaat ccaacttgaa gaaggttact ttggagttgg gtggtaaatc tcctaacatc
     1441 gtgttcaacg atgctaacat taagcaagct gtcgccaaca tcatcctcgg tatttactac
     1501 aactctggag aagtttgttg tgctggttcc agagtttatg ttcaatccgg tatttacgac
     1561 gagcttttgg ccgaattcaa gactgctgct gagaatgtca aggttggtaa cccattcgac
     1621 gaggacacct tccaaggtgc tcaaacctct cagcaacaat tggagaagat tttgggtttc
     1681 gttgagcgtg gtaagaagga cggtgctact ttgattactg gtggtggcag attaggtgac
     1741 aagggttact tcgtccagcc aactatcttc ggtgatgtta caccagagat ggagattgtc
     1801 aaggaagaga tctttggtcc tgttgtcact atcagcaagt ttgacaccat tgatgaggtt
     1861 gtcgaccttg ctaacgactc tcaatacggt cttgctgctg gtatccactc tgacgatatc
     1921 aacaaggtca ttgacgttgc tgctagaatc aagtccggta ccgtgtgggt caacacctac
     1981 aacgatttcc accaaatggt tccattcggt ggatttggcc aatccggtat tggtcgtgag
     2041 atgggtgttg aagctttgga aaactacacc caatacaagg ctatccgtgt caagatcaac
     2101 cacaagaacg agagtcgagc cgcggcggcc gccagctttc tagaacaaaa actcatctca
     2161 gaagaggatc tgaatagcgc cgtcgaccat catcatcatc atcattgagt ttgtagcctt
     2221 agacatgact gttcctcagt tcaagttggg cacttacgag aagaccggtc ttgctagatt
     2281 ctaatcaaga ggatgtcaga atgccatttg cctgagagat gcaggcttca tttttgatac
     2341 ttttttattt gtaacctata tagtatagga ttttttttgt cattttgttt cttctcgtac
     2401 gagcttgctc ctgatcagcc tatctcgcag ctgatgaata tcttgtggta ggggtttggg
     2461 aaaatcattc gagtttgatg tttttcttgg tatttcccac tcctcttcag agtacagaag
     2521 attaagtgag accttcgttt gtgcggatcc cccacacacc atagcttcaa aatgtttcta
     2581 ctcctttttt actcttccag attttctcgg actccgcgca tcgccgtacc acttcaaaac
     2641 acccaagcac agcatactaa atttcccctc tttcttcctc tagggtgtcg ttaattaccc
     2701 gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt tctttttctt cgtcgaaaaa
     2761 ggcaataaaa atttttatca cgtttctttt tcttgaaaat tttttttttt gatttttttc
     2821 tctttcgatg acctcccatt gatatttaag ttaataaacg gtcttcaatt tctcaagttt
     2881 cagtttcatt tttcttgttc tattacaact ttttttactt cttgctcatt agaaagaaag
     2941 catagcaatc taatctaagg gcggtgttga caattaatca tcggcatagt atatcggcat
     3001 agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt
     3061 gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc
     3121 ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat
     3181 cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg
     3241 cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc
     3301 cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga
     3361 cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg tccgacgcgg
     3421 cccgacgggt ccgaggcctc ggagatccgt cccccttttc ctttgtcgat atcatgtaat
     3481 tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag
     3541 gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa
     3601 gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta
     3661 acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgca
     3721 agctggagac caacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3781 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3841 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3901 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3961 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4021 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4081 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4141 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4201 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4261 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4321 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4381 aagaagatcc tttgatcttt tctacggggt ct
//
