LOCUS       sequence-437141-        4541 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pGAP-like vector encoding Aspergillus nidulans 
            4'-phosphopantetheinyl transferase NpgA under control of GAP 
            promoter, for yeast expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4541)
  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 4541)
  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: pNK1508
FEATURES             Location/Qualifiers
     source          1..4541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             242..1270
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTCYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      1276..1473
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     terminator      1537..1784
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1553..1571)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(1859..2447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1939..1958)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2538..3014
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     terminator      4134..4380
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     gene            join(4439..4541,1..1473)
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        join(4439..4541,1..241)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
ORIGIN
        1 aatcatttgc atccatacat tttgatggcc gcacggcgcg aagcaaaaat tacggctcct
       61 cgctgcggac ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg
      121 ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc actgaggttc ttctttcata
      181 tacttccttt taaaatcttg ctaggataca gttctcacat cacatccgaa cataaacaac
      241 catgggtaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
      301 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
      361 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
      421 agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga
      481 cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
      541 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat
      601 ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
      661 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
      721 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
      781 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
      841 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
      901 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
      961 ggcttgtatg gagcagcaga cgtgctactt cgagcggagg catccggagc ttgcaggatc
     1021 gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     1081 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     1141 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga
     1201 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
     1261 aaaggaataa tcagtactga caataaaaag attcttgttt tcaagaactt gtcatttgta
     1321 tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat ttatattttt
     1381 tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt aatatcatgc
     1441 gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta acgccgccat
     1501 ccagtgtcga aaacgagctc gaattcatcg atgatatcat gtaattagtt atgtcacgct
     1561 tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg
     1621 aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat
     1681 ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa
     1741 ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcaagctg gagactaaca
     1801 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1861 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1921 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1981 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2041 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2101 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2161 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2221 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2281 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2341 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2401 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2461 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2521 tgcatgagat cagatctttt ttgtagaaat gtcttggtgt cctcgtccaa tcaggtagcc
     2581 atctctgaaa tatctggctc cgttgcaact ccgaacgacc tgctggcaac gtaaaattct
     2641 ccggggtaaa acttaaatgt ggagtaatgg aaccagaaac gtctcttccc ttctctctcc
     2701 ttccaccgcc cgttaccgtc cctaggaaat tttactctgc tggagagctt cttctacggc
     2761 ccccttgcag caatgctctt cccagcatta cgttgcgggt aaaacggagg tcgtgtaccc
     2821 gacctagcag cccagggatg gaaaagtccc ggccgtcgct ggcaataata gcgggcggac
     2881 gcatgtcatg agattattgg aaaccaccag aatcgaatat aaaaggcgaa cacctttccc
     2941 aattttggtt tctcctgacc caaagacttt aaatttaatt tatttgtccc tatttcaatc
     3001 aattgaacaa ctataccgcg cacagaactc ctactctact agtaaaaaaa atggtccaag
     3061 acacctcaag cgctagtacc agtccaattt tgacaagatg gtatatagat actagacctt
     3121 tgactgcttc taccgcagcc ttgccactgt tggaaactct gcagcctgca gaccagataa
     3181 gtgtccaaaa gtactaccat ctgaaggata agcatatgtc tctcgcatcc aatcttctca
     3241 agtacttgtt tgttcacagg aattgccgaa tcccatggtc tagcattgta atatcaagaa
     3301 ctccagatcc ccatagaagg ccctgttata taccaccctc tggttctcaa gaggacagct
     3361 ttaaggatgg ctacactgga attaatgtcg agttcaacgt atcccatcaa gcaagcatgg
     3421 tggcaatagc aggcactgca tttacaccca actctggagg tgattccaaa cttaagccag
     3481 aggttggcat agacattact tgcgtgaatg agcgacaggg gcgaaacggt gaggagcgat
     3541 cccttgaaag tttgaggcag tacatagata ttttctcaga ggttttttcc accgcagaga
     3601 tggcaaacat cagaagattg gatggggtgt catcaagctc cttgagcgcc gataggttgg
     3661 tggactatgg ttaccgattg ttttacactt attgggctct taaggaagca tacatcaaga
     3721 tgacagggga agccctgctg gcaccatggc tcagggagtt ggaatttagt aatgtcgttg
     3781 ccccagcagc agtggccgaa tccggagatt ccgccggaga ctttggagag ccttataccg
     3841 gggtacgaac aaccctttac aaaaacttgg tcgaggatgt taggatagag gtggctgctc
     3901 tgggaggtga ctacttgttc gccactgctg cacgaggggg cggtataggt gcctccagcc
     3961 gaccaggggg aggccctgat ggctcaggaa ttaggagtca agatccctgg cgaccattta
     4021 aaaagctcga catagagaga gacatacagc cttgcgccac tggagtctgt aattgtctgt
     4081 ctcgaggtta atctaagctg agctacacga atctatagga accgggaact tagtcaagag
     4141 gatgtcagaa tgccatttgc ctgagagatg caggcttcat ttttgatact tttttatttg
     4201 taacctatat agtataggat tttttttgtc attttgtttc ttctcgtacg agcttgctcc
     4261 tgatcagcct atctcgcagc tgatgaatat cttgtggtag gggtttggga aaatcattcg
     4321 agtttgatgt ttttcttggt atttcccact cctcttcaga gtacagaaga ttaagtgaga
     4381 cattcgtttg tgcggatctg tttagcttgc ctcgtccccg ccgggtcacc cggccagcga
     4441 catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg
     4501 tgactgtcgc ccgtacattt agcccataca tccccatgta t
//
