LOCUS       sequence-292149-        6754 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of COX4-DuDre as an additional copy in genome, use 
            auxotrophic marker TRP1(Kluyveromyces lactis). Marker for yeast 
            mitochondria..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6754)
  AUTHORS   Zhu J, Zhang ZT, Tang SW, Zhao BS, Li H, Song JZ, Li D, Xie Z
  TITLE     A Validated Set of Fluorescent-Protein-Based Markers for Major 
            Organelles in Yeast (Saccharomyces cerevisiae).
  JOURNAL   MBio. 2019 Sep 3;10(5). pii: mBio.01691-19. doi: 
            10.1128/mBio.01691-19.
  PUBMED    31481383
REFERENCE   2  (bases 1 to 6754)
  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: "MBio."; 
            date: "2019-09-3"; volume: "10(5). pii: mBio.01691-19. doi"; pages: 
            " 10.1128/mBio.01691-19"
COMMENT     Sequence Label: ZJOM64
FEATURES             Location/Qualifiers
     source          1..6754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1021..1086
                     /codon_start=1
                     /product="mitochondrial presequence of yeast cytochrome c 
                     oxidase subunit IV (Hurt et al., 1984)"
                     /label=pre-CoxIV
                     /translation="MLSLRQSIRFFKPATRTLCSSR"
     terminator      3030..3217
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3085..3109)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    3269..3302
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4305..4338
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(4391..4410)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4392..4410)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4497..4514)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4668..5256)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4748..4767)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5427..6218,6219..6287))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6050..6069
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6288..6392)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6460..6478
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6516..6538)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6638..6657
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(6738..6754,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(6738..6754,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcctattgga ttggcccaat tgctgccaaa tttggtgaat
       61 atggtggtga tgtcgcaatg tggttaagta tggcgttctc aggtgttgta taccctcctt
      121 gtagatactt agagctacgt aaatttggtc gttaattgaa atgacataag tatatattaa
      181 aactacggtg cagcggcaaa ctttagttta gataatattg gcaggtagat tcatggtatg
      241 ttaaattacg ctcagggagg atattttttt gtacacatta tcttttcaca tgaaaatcta
      301 agcctggtgg acaattgaaa tcatcgagtg atatactgac actttgaaca gtagcacaaa
      361 tcgtttaatt tgctgtcacg atctaagttt tctgtcagct gactgtccac caataagatc
      421 agcggattcc aaaggcgtcc ttcgaaaaag cgatatctcc catcttggtt agtagcaaat
      481 aaagggaaga gaaaataaga cacactcaag gataaaatac ttcgcttctt ccaagtataa
      541 acacaggaaa gaaagatata cgaagattag aatttttttc atacttttta caaaacattt
      601 ttttgtactt aggcacgcta attgcgtaaa aattagcata aacaggaaat aagattattt
      661 tgaaatgtat gttataatcc tcttattgaa aaatacatga gatgtaggag aagaactacc
      721 aggaaagaag atatttaaca gatcgaacga aacggaaaag aactaagaga aaacaaatct
      781 ggatttgact ttaagaaaaa atttatttgc aagtttaact cgttccctgc ttttctattt
      841 ggcttgagac gataattgca taattgtcca aattgtccat catataatga agaattcgat
      901 gcgaaatatt ccctgtcact ttcctttagt caacgtattc ttctccctga agaaacagta
      961 tactaacaat actcacccat ttcgattttg atgttgccat acaaatagat aacaagcaca
     1021 atgctttcac tacgtcaatc tataagattt ttcaagccag ccacaagaac tttgtgtagc
     1081 tctagatatc tgcttcagca aaaacccgtg gtgaaaactg cccaaaactt agcagaagtt
     1141 aatggtccag aaactttgat tggtcctggt gctaaagagg gtaccgttcc aacagaccta
     1201 gatcaagaaa ctggtttagc taggttagaa ttattgggta aattagaggg tatcgatgtt
     1261 ttcgacacca aaccattaga ttcgtccagg aagggtacca tgaaagatcc gatcatcatt
     1321 gaatcttatg atgattatcg ttatgtcggt tgtacgggtt ctcccgctgg ttcacatact
     1381 attatgtggt taaaaccaac tgttaacgaa gtcgccagat gctgggaatg tggttctgtt
     1441 tacaaactaa accctgttgg tgttccaaat gatgaccacc atcacgcttc gtacgctgca
     1501 ggtcgtggga ttcctgggtt aattaacatc gctagcatgg tttccaaagg tgaagagaac
     1561 aatatggctg tcatcaagcc cttcatgaga ttcaaggtgc acatggaggg ctccgtgaac
     1621 ggccacgagt tcgagatcga gggcgagggc gagggcaagc cctacgaggg cacccagacc
     1681 gccaagctgc aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc
     1741 cagttccagt acggctccaa ggtgtacgtg aagcaccccg ccgacatccc cgactacaag
     1801 aagctgtcct tccccgaggg cttcaagtgg gagagagtga tgaacttcga ggacggcggc
     1861 gtggtgaccg tgacccagga ctcctccctg caggacggca ccttcatcta ccacgtgaag
     1921 ttcatcggcg tgaacttccc ctccgacggc cccgtaatgc agaagaagac tctgggctgg
     1981 gagccctcca ccgagagact gtaccccaga gacggcgtgc tgaagggcga gatccacaag
     2041 gcgctgaagc tgaagggcgg cggccactac ctggtggagt tcaagtcaat ctacatggcc
     2101 aagaagcccg tgaagctgcc cggctactac tacgtggact ccaagctgga catcacctcc
     2161 cacaacgagg actacaccgt ggtggagcag tacgagagag ccgaggccag acaccacctg
     2221 ttcttaattc cagttccaac gcgtgatctt gagaagccat tcaacgtcat caagcccttc
     2281 atgagattca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     2341 gagggcgagg gcaagcccta cgagggcacc cagaccgcca agctgcaggt gaccaagggc
     2401 ggccccctgc ccttcgcctg ggacatcctg tccccccagt tccagtacgg ctccaaggtg
     2461 tacgtgaagc accccgccga catccccgac tacaagaagc tgtccttccc cgagggcttc
     2521 aagtgggaga gagtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     2581 tccctgcagg acggcacctt catctaccac gtgaagttca tcggcgtgaa cttcccctcc
     2641 gacggccccg taatgcagaa gaagactctg ggctgggagc cctccaccga gagactgtac
     2701 cccagagacg gcgtgctgaa gggcgagatc cacaaggcgc tgaagctgaa gggcggcggc
     2761 cactacctgg tggagttcaa gtcaatctac atggccaaga agcccgtgaa gctgcccggc
     2821 tactactacg tggactccaa gctggacatc acctcccaca acgaggacta caccgtggtg
     2881 gagcagtacg agagagccga ggccagacac cacctgttct taattgtcgt tctgaacaga
     2941 gctgagactc cattgccatt ggatccaacc ggtaaggtca aggctgagct ggatactagg
     3001 cctaggtgag gcgcgccact tctaaataag cgaatttctt atgatttatg atttttatta
     3061 ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa
     3121 aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat
     3181 agtatgaggt cgctcttatt gaccacacct ctaccggcag atccgctagg gataacaggg
     3241 taatatctgc aggtcgacaa cccttaatat aacttcgtat aatgtatgct atacgaagtt
     3301 attaggtcta gagcggccgc ggaacgatca ttcactatat atatatcaat ttatatatac
     3361 gtatgtgtaa ttgaagaaag atacgttttt tcctctattg agaggcctgc tggatgaata
     3421 gctttacctt ttctaaatcc ttgataccat cagtctctac tcctccactt acatcgacac
     3481 cgatcgcatt tggtaacata ttaatggcaa cagaaacgtt atcaggattc aatccaccag
     3541 cgataatgaa ttttatctcg ggatgacttg cagaccaact ggaaattgca ctccaattca
     3601 atttctcacc agtgccacct tcaccagaat cgaacaacgt cagcacattg tctacgtgtt
     3661 catacaggtc cagtagtaat tcacaatcct gtgggaactg gaacctctta atgattggaa
     3721 ttgaagatgg gatcaaagat ctgtattctt taatatcttc atctccatgt aattgtatca
     3781 catctagatt atattcgtgg tacagttgaa ggacatcatc aacggactga tttctaaaca
     3841 ccccgactag tttagtacct ttcacgttct cttgttggtg aactgcagtt gaaatacctt
     3901 tcgcaacaga tgagtcaatg gttcttttcc taccgggaac acaaatgata cctaagtaat
     3961 cagcaccatc atccacagca gtctttgcag cttcaacggt ttgcaaacca cacactttaa
     4021 cgagcatcac agtagatttc gcaagagggt agctcgctca gagtacccaa gtaaatgatt
     4081 agtaaactga tgtttgatag ttcaattttt caatgaaata accttatatt aaaattgata
     4141 ttactattat acaaaaataa agaataaagg atttgagttt atacataaaa taccattatt
     4201 atttgttcag tgagagatac tggggtatat gggatgtgtg tagtgatacc atgcaatcat
     4261 gtatcaaaca tgggcccggt accgagctct cgagaaccct taatataact tcgtataatg
     4321 tatgctatac gaagttatta ggtgatatca gatccactag tggcctatgc ggccgcggat
     4381 ctgccggtct ccctatagtg agtcgtatta atttcgataa gccaggttaa cctgcattaa
     4441 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4501 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4561 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4621 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4681 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4741 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4801 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4861 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4921 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4981 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5041 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5101 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5161 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5221 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5281 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5341 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5401 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5461 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5521 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5581 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5641 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5701 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5761 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5821 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5881 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5941 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6001 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6061 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6121 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6181 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6241 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6301 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6361 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6421 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     6481 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     6541 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     6601 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     6661 agagtgcacc atatggacat attgtcgtta gaacgcggct acaattaata cataacctta
     6721 tgtatcatac acatacgatt taggtgacac tata
//
