LOCUS       sequence-438221-        4589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for expression HmS in yeast (P.pastoris).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4589)
  AUTHORS   Palkina KA, Karataeva TA, Perfilov MM, Fakhranurova LI, Markina NM, 
            Somermeyer LG, Garcia-Perez E, Vazquez-Vilar M, Rodriguez-Rodriguez 
            M, Vazquez-Vilriales V, Shakhova ES, Mitiouchkina T, Belozerova OA, 
            Kovalchuk SI, Alekberova A, Malyshevskaia AK, Bugaeva EN, Guglya EB,
            Balakireva A, Sytov N, Bezlikhotnova A, Boldyreva DI, Babenko VV, 
            Kondrashov FA, Choob VV, Orzaez D, Yampolsky IV, Mishin AS, 
            Sarkisyan KS
  TITLE     A hybrid pathway for self-sustained luminescence.
  JOURNAL   Sci Adv. 2024 Mar 8;10(10):eadk1992. doi: 10.1126/sciadv.adk1992. 
            Epub 2024 Mar 8.
  PUBMED    38457503
REFERENCE   2  (bases 1 to 4589)
  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: "Sci Adv."; 
            date: "2024-03-8"; volume: "10(10):eadk1992. doi"; pages: " 
            10.1126/sciadv.adk1992. Epub 2024 Mar 8"
COMMENT     Sequence Label: pNK7181
FEATURES             Location/Qualifiers
     source          1..4589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        452..928
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             2138..2155
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2201..2230
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2246..2263
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2343..2589
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2604..3017
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3024..3071
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3090..3464
                     /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      3530..3777
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3546..3564)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3852..4440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3932..3951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acctaaacgc ttaatgcctc agacgtttta atgcctctca acacctccaa ggttgctttc
       61 ttgagcatgc ctactaggaa ctttaacgaa ctgtggggtt gcagacagtt tcaggcgtgt
      121 cccgaccaat atggcctact agactctctg aaaaatcaca gttttccagt agttccgatc
      181 aaattaccat cgaaatggtc ccataaacgg acatttgaca tccgttcctg aattatagtc
      241 ttccaccgtg gatcatggtg ttcctttttt tcccaaagaa tatcagcatc ccttaactac
      301 gttaggtcag tgatgacaat ggaccaaatt gttgcaaggt ttttcttttt ctttcatcgg
      361 cacatttcag cctcacatgc gactattatc gatcaatgaa atccatcaag attgaaatct
      421 taaaattgcc cctttcactt gacaggatcc ttttttgtag aaatgtcttg gtgtcctcgt
      481 ccaatcaggt agccatctct gaaatatctg gctccgttgc aactccgaac gacctgctgg
      541 caacgtaaaa ttctccgggg taaaacttaa atgtggagta atggaaccag aaacgtctct
      601 tcccttctct ctccttccac cgcccgttac cgtccctagg aaattttact ctgctggaga
      661 gcttcttcta cggccccctt gcagcaatgc tcttcccagc attacgttgc gggtaaaacg
      721 gaggtcgtgt acccgaccta gcagcccagg gatggaaaag tcccggccgt cgctggcaat
      781 aatagcgggc ggacgcatgt catgagatta ttggaaacca ccagaatcga atataaaagg
      841 cgaacacctt tcccaatttt ggtttctcct gacccaaaga ctttaaattt aatttatttg
      901 tccctatttc aatcaattga acaactatca attcgaaacg atggctacca aaagtgtcgc
      961 cgttgaagaa atgtgcaaag cccaaaaagc cggaggccct gcaacaatcc ttgcaattgg
     1021 tactgctgtg cccagtaact gctactatca gagtgaatac cctgatttct acttccgtgt
     1081 cacgaaaagt gatcacctta cggacctgaa aagtaagttt aaacgtatgt gcgaacgttc
     1141 atccatcaaa aagcgttata tgcacctgac agaggagatt cttgaagaaa atcctaatat
     1201 gtgcacgttt gcagctccat ctatcgatgg ccgtcaggac attgtagtta aggagatccc
     1261 taagttggct aaggaggcag caagtaaagc tatcaaggaa tggggccaac ctaagtcaaa
     1321 tatcacccat cttgtctttt gtacaacttc aggtgtggac atgcccggtt gtgattatca
     1381 attgacgcgt cttttgggac ttcgtccatc tattaaacgt cttatgatgt atcagcaggg
     1441 ctgccatgct ggtggaaccg gcttgcgttt ggccaaagac cttgccgaaa ataacaaggg
     1501 agctcgtgta ttggtggttt gttctgagat gacagtgatt aactttcgtg gccccagtga
     1561 ggcccacatg gattcccttg tcggacaatc tcttttcggc gacggcgcct cagccgtaat
     1621 cgtaggttca gatcccgatc tgtcaacaga gcatccactt taccagatta tgtccgcctc
     1681 ccaaatcatt gtagccgact ccgaaggcgc tattgatggt cacttgcgtc aagagggtct
     1741 tacctttcat ctgcgtaagg atgttccttc tcttgtctcc gataacatcg agaatactct
     1801 ggtggaagca tttactccca tcctgatgga cagtatcgac tcaatcattg attggaactc
     1861 tatcttttgg attgctcacc caggtggtcc tgccatcctg aatcaggttc aagcaaaagt
     1921 aggactgaag gaggaaaaac tgcgtgtgag tcgtcacatt ttgtcagaat atggcaatat
     1981 gtcatccgca tgcgtctttt ttatcatgga tgagatgcgt aaacgttcca tggaagaggg
     2041 aaaaggaact accggcgagg gccttgagtg gggagtgctt tttggcttcg gtcctggttt
     2101 tacggtcgaa accattgtgt tgcacagtgt gcccattcat caccaccacc atcactaaac
     2161 tagttcggta cctcgagccg cggcggccgc cagctttcta gaacaaaaac tcatctcaga
     2221 agaggatctg aatagcgccg tcgaccatca tcatcatcat cattgagttt gtagccttag
     2281 acatgactgt tcctcagttc aagttgggca cttacgagaa gaccggtctt gctagattct
     2341 aatcaagagg atgtcagaat gccatttgcc tgagagatgc aggcttcatt tttgatactt
     2401 ttttatttgt aacctatata gtataggatt ttttttgtca ttttgtttct tctcgtacga
     2461 gcttgctcct gatcagccta tctcgcagct gatgaatatc ttgtggtagg ggtttgggaa
     2521 aatcattcga gtttgatgtt tttcttggta tttcccactc ctcttcagag tacagaagat
     2581 taagtgagac cttcgtttgt gcggatcccc cacacaccat agcttcaaaa tgtttctact
     2641 ccttttttac tcttccagat tttctcggac tccgcgcatc gccgtaccac ttcaaaacac
     2701 ccaagcacag catactaaat ttcccctctt tcttcctcta gggtgtcgtt aattacccgt
     2761 actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg
     2821 caataaaaat ttttatcacg tttctttttc ttgaaaattt ttttttttga tttttttctc
     2881 tttcgatgac ctcccattga tatttaagtt aataaacggt cttcaatttc tcaagtttca
     2941 gtttcatttt tcttgttcta ttacaacttt ttttacttct tgctcattag aaagaaagca
     3001 tagcaatcta atctaagggc ggtgttgaca attaatcatc ggcatagtat atcggcatag
     3061 tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc
     3121 tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc
     3181 gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca
     3241 gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc
     3301 tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg
     3361 ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc
     3421 cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtc cgacgcggcc
     3481 cgacgggtcc gaggcctcgg agatccgtcc cccttttcct ttgtcgatat catgtaatta
     3541 gttatgtcac gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga
     3601 gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga
     3661 acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac
     3721 attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcaag
     3781 ctggagacca acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3841 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3901 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3961 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4021 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4081 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4141 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4201 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4261 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4321 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4381 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4441 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4501 gggattttgg tcatgagatc agatctctgc tactctggtc ccaagtgaac caccttttgg
     4561 accctattga ccggacctta acttgccaa
//
