LOCUS       sequence-441226-        8428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pGAP-like vector encoding hispidin-synthase from Mycena citricolor 
            under control of GAP promoter, for yeast expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8428)
  AUTHORS   Shakhova ES, Karataeva TA, Markina NM, Mitiouchkina T, Palkina KA, 
            Perfilov MM, Wood MG, Hoang TT, Hall MP, Fakhranurova LI, Alekberova
            AE, Malyshevskaia AK, Gorbachev DA, Bugaeva EN, Pletneva LK, Babenko
            VV, Boldyreva DI, Gorokhovatsky AY, Balakireva AV, Gao F, Choob VV, 
            Encell LP, Wood KV, Yampolsky IV, Sarkisyan KS, Mishin AS
  TITLE     An improved pathway for autonomous bioluminescence imaging in 
            eukaryotes.
  JOURNAL   Nat Methods. 2024 Mar;21(3):406-410. doi: 
            10.1038/s41592-023-02152-y. Epub 2024 Jan 22.
  PUBMED    38253843
REFERENCE   2  (bases 1 to 8428)
  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"; doi: 
            "10.1038/s41592-023-02152-y"; journalName: "Nat Methods"; date: 
            "2024-03"; volume: "21"; issue: "3"; pages: "406-410"
COMMENT     Sequence Label: pNK5867
FEATURES             Location/Qualifiers
     source          1..8428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1007..1483
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             6601..6630
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             6646..6663
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      6743..6988
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        7004..7417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        7424..7471
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7490..7864
                     /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      7930..8177
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(7946..7964)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(join(8246..8428,1..406))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8326..8345)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
       61 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
      121 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
      181 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
      241 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
      301 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
      361 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
      421 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
      481 gagatcagat ctctgctact ctggtcccaa gtgaaccacc ttttggaccc tattgaccgg
      541 accttaactt gccaaaccta aacgcttaat gcctcagacg ttttaatgcc tctcaacacc
      601 tccaaggttg ctttcttgag catgcctact aggaacttta acgaactgtg gggttgtaga
      661 cagtttcagg cgtgtcccga ccaatatggc ctactagact ctctgaaaaa tcacagtttt
      721 ccagtagttc cgatcaaatt accatcgaaa tggtcccata aacggacatt tgacatccgt
      781 tcctgaatta tagtcttcca ccgtggatca tggtgttcct ttttttccca aagaatatca
      841 gcatccctta actacgttag gtcagtgatg acaatggacc aaattgttgc aaggtttttc
      901 tttttctttc atcggcacat ttcagcctca catgcgacta ttatcgatca atgaaatcca
      961 tcaagattga aatcttaaaa ttgccccttt cacttgacag gatccttttt tgtagaaatg
     1021 tcttggtgtc ctcgtccaat caggtagcca tctctgaaat atctggctcc gttgcaactc
     1081 cgaacgacct gctggcaacg taaaattctc cggggtaaaa cttaaatgtg gagtaatgga
     1141 accagaaacg tctcttccct tctctctcct tccaccgccc gttaccgtcc ctaggaaatt
     1201 ttactctgct ggagagcttc ttctacggcc cccttgcagc aatgctcttc ccagcattac
     1261 gttgcgggta aaacggaggt cgtgtacccg acctagcagc ccagggatgg aaaagtcccg
     1321 gccgtcgctg gcaataatag cgggcggacg catgtcatga gattattgga aaccaccaga
     1381 atcgaatata aaaggcgaac acctttccca attttggttt ctcctgaccc aaagacttta
     1441 aatttaattt atttgtccct atttcaatca attgaacaac tatcaattcg aagaggaatt
     1501 caagcttcat atggctagca aatgtcagcc tcttcatctt attctgaatt ggagtctcca
     1561 acttctcttc ttgacgtgtt cgtgcacgcc gctcgggacc ctcacaccgc atcaagaagg
     1621 gtgttggagt gcggatctga cacatggaca tacgccgcct tggatgccgt gtctgatggt
     1681 atagccaggg agttggcccc tttcggtttg gccccaaagg tcgctgttgt ctctgagaac
     1741 catccatttg tgttcgcttt gttgttcgca gtctggaagt tgggtggaac tttcatccca
     1801 atcgacgcac atgtcccttt tgccatgttg acaggtatgg tgaacatcgt gaagccaaca
     1861 tgtctttacc ttcctgcctc agccacatca aacatatctc ttgtgaaggc cttcgacata
     1921 agaaccgtcg tgtttggaca caaggagaat tcaatgcaag ctttgttcga caagtactca
     1981 ttgcacgcag ccccattgca cgctgcacca ttgcattacg cccctccatc agccgaccat
     2041 gcctgtcttt atcttttcac ttcttcagcc tcttctacca aaaacttgaa ggccgtccct
     2101 cttacccata cacttgtctt gagaggttgt caatctaaac ttgcttggtg gaggagggtc
     2161 caaccaggaa agaatcttga cgctatcaga atccttggat gggcaccatg gtcccacgtt
     2221 cttgcctata tgcaggatat cggaacagct acacttctta acgccggttg ctacgtcttt
     2281 gcaaccatcc cttcttcata ccctcagctt cctaccacta ccccagtcga tttgaccact
     2341 tcacttatcg acgctttggt caccagaaga gtcgcagcat tcgcctgtgt ccctttcgtc
     2401 cttgccaacc ttaaagccgc ttgccagtca aaccacccag ccaggtcaca gttgcttgag
     2461 gcccttcaaa agaccatcat gttggagtgc ggtggtgccg tgttggacga cgctactgtg
     2521 gactgggccg agaggaacgg tatcaggatc ttcaccggaa tcggtatgac cgagactggt
     2581 ggagccgttt tcgtcggttt ggcctctgaa tctagaagag gatttttgcc tgaaggtttg
     2641 cttggtgatg catcattctc tatctcatct gacactgacg ctcttgacga aggtgaactt
     2701 gtcgtgaaat caaaattgat cgcaccagga tatgtcggat acgacgacgg agcccactct
     2761 gtggactgcg acggatgggt cacattcagg acaggagata ggtataggca gacccagcca
     2821 gacggaaggt ttacctggtt gggaaggatc acagacttca ttcagatggt gtctggtgag
     2881 tacttggatc caaggccatt ggaggagtct cttagagcct ctccattgat cgccaacgcc
     2941 tgcatcgtgg gtgacgcctt cctatcttct gcctcaactt caatcttggc catcatcgag
     3001 ttggcaaccc ctgatcttgc acacaccgcc tcaatcaggg cacagcttgc aagagtcctt
     3061 gcccctctta acagggattt gcctccacca cttaggatcg cagcatcttc aatccttgtt
     3121 ttggacggaa tgagaaagat ccctaagaca aagaagggtg acatcttcag aaaaaagctt
     3181 gaggacacat tcggtgctga gtttgagcag atgttgagaa ccgagaaagt cggattgggt
     3241 gacttggctg atgtggatgc cggtatcact agaatcatcg gaaacttgtt gggaatctca
     3301 gacgacgagc ttctttctac catgtctttt gctgagttgg gtatgacctc tcttttggcc
     3361 gtcaagatcg catcagaatt gaacaagttt ttggacggta gggctgtgtt gccaacaaac
     3421 atctgttaca tacactttga cgtgccttca ttgacttcat cagtcagaga aaggctttct
     3481 tctgcaccat catctatcac atctgccgcc tctgagcctg cagcctcttc accaggaacc
     3541 cctagggccg acgaaatcgt gatcgtggga aaggccttca gacttcctga cggtgtgaac
     3601 gacgacgccg ctttgtggga cgtccttacc ggtgagtctg catctatcat caaggacatc
     3661 ccagccgaca gatgggacca cgcctctttc tacccaaagg acatccactt tgggagagct
     3721 ggtcttgtgg atgtcgctag attcgactac ggttttttcg gaatgaccgc ttctgaggca
     3781 tattctcttt caccaaccat gaggttggct cttgaagtcg cttatgaggc cttggaggac
     3841 gcaaacattc ctttcagagc cgtcaagggt tcaaggatgg gtgtgttcgt cgctgtgaaa
     3901 gacgatggat tcgaaacctt gttgcatgca ggacagggtt atgacgctta cacaagattc
     3961 tatggaactg gaagggctcc ttcaaccgcc tctggtagaa tcaactattt gcttgatttg
     4021 cacggacctt caatcaccgt cgacacagcc tgctcaggag gaatcgtgtg catcgaccag
     4081 gctgtcacct atcttcaatc aggagctgca gagactgcca tcgtgtgttc atctaacaca
     4141 cactgctggc caggatcttt catgttcttg accgctcaag gaatggcctc acctaacggt
     4201 agatgcgcat ctttcacatc agacgccgac ggatacgctc cttctgaggg agctgtgggt
     4261 ttcgtgctta agaccaggtc tgccgctgtg agggacggag atagaatact tgctactatc
     4321 agggccaccg agatcggaca caatggaaga tctcagggac ttgctgctcc taatgtcaga
     4381 tctcaggcag ctgctcatag agccgtgttg cgtagagcaa ggttggaccc atctgagatt
     4441 gacttcatcg aagcacatgg tactggaacc acattgggag atctttgcga agtgcagggt
     4501 atcaacgact cttttgtctc tcctaagaaa agagccaacc ctcttgtagt ctctgcatct
     4561 aagtcaacca tcggtcacac cgagccatct gctggtttgg tcggaatcct ttcagctctt
     4621 atgtcattcg agaagaggat cgtgcctaga ttggcatatc ttactgagtc taatgtgaac
     4681 ccagcattgg acgcatcagt cgttccattg cactttccta caaagcacat cgagcttaga
     4741 gctgatgtgc cttacaaggc cgtggtgatg tcttacggat tcgcaggtac ccttgccgac
     4801 atcgtccttg agtcagaggt tccacaacca actccagccg tcgctcagga cacagccggt
     4861 caacagccaa tgcttttcgt gttgtcagcc aaaaccccaa gggcacttgc agcctacatc
     4921 gagctttacc ttggtttcct taggcacgcc gaccctggac ttttcgcaag gatctgctac
     4981 accgcatgtg tcgcaaggga gcactacaag cacagggtcg cctgcgttgc aacagacttg
     5041 gtcgacctta tcgctcagct tgagactagg cttgtgcaga ctgcctatgc aggaggagga
     5101 ggtgccaggg ccgctaggac cggtcctctt gtgtttgctt tttctggaca gggtactcag
     5161 ttccctgcta tggcagcacc acttgctagg agatacgcaa gattcggaga gatcgtgggt
     5221 ggttgtgcta ggatggcaag agagctttca ggtttcccag tggatggtat tcttcttggt
     5281 gacgatgtga cacctgtgaa ggacaactct gctgcagctg aggtgcactc tgaggtggac
     5341 cagatctgca tatttgtgta ccagtatgcc atgtgtagat ggttgggtga gcttggtgtg
     5401 gagccaaagg ctgccatagg tcactctttg ggagagatca cagcagctgt catcgcagga
     5461 gcacttcctt tcgaagccgc cttggatctt gtcgtcacta gagccaggtt gcttaagcca
     5521 tgcgccgagc aaccttctgg aatggcagcc ttggcctgca ccccagacgt cgcatcaaag
     5581 ttgacacttg gtgcttcagt gtctgtgtca gtctacaacg gacctcagtc tatctgcctt
     5641 tctggtgctt ctgccgagct tgacgatgcc gtgagggccg ctaaggccag gaacatcaag
     5701 gcaactagac ttcaggtcga ccagggtttc cactcacctt gcgtcgacgc cgcagtgcca
     5761 ggtttgcagg cttggtgcgc agctcacagg gcatcagctg ctccattgaa gatgcctctt
     5821 tactctaccg tgagagggga cgtcgttcca aagggagccg cccttgatcc agagcactgg
     5881 gtggcccacg ctaggaaccc tgtgttgttc gcccagacag cccaggctct taaagagtca
     5941 cttccacatg gaatcacttt ggacgtcggt cctcaggccg tggcttggtc tttgcttctt
     6001 ttgaacggtc tttcagctac cagaacagtc gctgccggag ccaagaaggg agcagaccag
     6061 gagagggccc ttttgggtgc tcttggagcc ttgttcgagc agcacaaggt cactcctgac
     6121 tttggtagac tttacgctcc tttggaaaag acaaggatcc caacctaccc atttgagaga
     6181 gctagatgct acccaacctt cataccatca aggttcgccc acggtgctgc tgctgctgcc
     6241 aagacagatg gtgaggtcct ttctgccgaa gaagaaaatg tggcacctgt gggattgttg
     6301 acaaaggagg atttgagggc agctttggtc gcttgtttga gggccacatt ggagcttagg
     6361 cctgacgagg agcttgacga agcagagcct ttgaccgtgc acggagtgga ctctatcgga
     6421 tttgcaaagc ttaggaagca cgtcgaggac aggtggggtc ttgacatccc agtcgtgtac
     6481 tggtcagacg ccttcaccgt cggagagatg ttgggtaact tggtcggaca gtatgacgtg
     6541 gtgtctactg ctgccaggta gttaactagc ttagctaaat aagcggccgc cagctttcta
     6601 gaacaaaaac tcatctcaga agaggatctg aatagcgccg tcgaccatca tcatcatcat
     6661 cattgagttt gtagccttag acatgactgt tcctcagttc aagttgggca cttacgagaa
     6721 gaccggtctt gctagattct aatcaagagg atgtcagaat gccatttgcc tgagagatgc
     6781 aggcttcatt tttgatactt ttttatttgt aacctatata gtataggatt tttttgtcat
     6841 tttgtttctt ctcgtacgag cttgctcctg atcagcctat ctcgcagctg atgaatatct
     6901 tgtggtaggg gtttgggaaa atcattcgag tttgatgttt ttcttggtat ttcccactcc
     6961 tcttcagagt acagaagatt aagtgagaac cttcgtttgt gcggatcccc cacacaccat
     7021 agcttcaaaa tgtttctact ccttttttac tcttccagat tttctcggac tccgcgcatc
     7081 gccgtaccac ttcaaaacac ccaagcacag catactaaat ttcccctctt tcttcctcta
     7141 gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc
     7201 tttttcttcg tcgaaaaagg caataaaaat ttttatcacg tttctttttc ttgaaaattt
     7261 ttttttttga tttttttctc tttcgacgac ctcccattga tatttaagtt aataaacggt
     7321 cttcaatttc tcaagtttca gtttcatttt tcttgttcta ttacaacttt ttttacttct
     7381 tgctcattag aaagaaagca tagcaatcta atctaagggc ggtgttgaca attaatcatc
     7441 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt
     7501 gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac
     7561 cgaccggctc gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga
     7621 cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc
     7681 ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac
     7741 gaacttccgg gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg
     7801 ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga
     7861 ctgacacgtc cgacgcggcc cgacgggtcc gaggcctcgg agatccgtcc cccttttcct
     7921 ttgtcgatat catgtaatta gttatgtcac gcttacattc acgccctccc cccacatccg
     7981 ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt tattttttta
     8041 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     8101 gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct
     8161 cgaaggcttt aatttgcaag ctgaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     8221 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     8281 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     8341 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     8401 tgtccgcctt tctcccttcg ggaagcgt
//
