LOCUS       sequence-271502-        6007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of COX4-mTagBFP2 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 6007)
  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 6007)
  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: ZJOM140
FEATURES             Location/Qualifiers
     source          1..6007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     173..191
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(229..251)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     351..370
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        451..469
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     451..468
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1539..1604
                     /codon_start=1
                     /product="mitochondrial presequence of yeast cytochrome c 
                     oxidase subunit IV (Hurt et al., 1984)"
                     /label=pre-CoxIV
                     /translation="MLSLRQSIRFFKPATRTLCSSR"
     terminator      2750..2937
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2805..2829)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    2989..3022
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4025..4058
                     /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(4111..4130)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4112..4130)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4217..4234)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4388..4976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4468..4487)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5147..5938,5939..6007))
                     /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     5770..5789
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
      181 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
      241 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
      301 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
      361 gcagattgta ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta
      421 atacataacc ttatgtatca tacacatacg atttaggtga cactatagaa cgcggccgcc
      481 agctgaagct tcgtacgctg caggtcgtgg gattcctggg ttaattaaca tcgctagcgc
      541 ctattggatt ggcccaattg ctgccaaatt tggtgaatat ggtggtgatg tcgcaatgtg
      601 gttaagtatg gcgttctcag gtgttgtata ccctccttgt agatacttag agctacgtaa
      661 atttggtcgt taattgaaat gacataagta tatattaaaa ctacggtgca gcggcaaact
      721 ttagtttaga taatattggc aggtagattc atggtatgtt aaattacgct cagggaggat
      781 atttttttgt acacattatc ttttcacatg aaaatctaag cctggtggac aattgaaatc
      841 atcgagtgat atactgacac tttgaacagt agcacaaatc gtttaatttg ctgtcacgat
      901 ctaagttttc tgtcagctga ctgtccacca ataagatcag cggattccaa aggcgtcctt
      961 cgaaaaagcg atatctccca tcttggttag tagcaaataa agggaagaga aaataagaca
     1021 cactcaagga taaaatactt cgcttcttcc aagtataaac acaggaaaga aagatatacg
     1081 aagattagaa tttttttcat actttttaca aaacattttt ttgtacttag gcacgctaat
     1141 tgcgtaaaaa ttagcataaa caggaaataa gattattttg aaatgtatgt tataatcctc
     1201 ttattgaaaa atacatgaga tgtaggagaa gaactaccag gaaagaagat atttaacaga
     1261 tcgaacgaaa cggaaaagaa ctaagagaaa acaaatctgg atttgacttt aagaaaaaat
     1321 ttatttgcaa gtttaactcg ttccctgctt ttctatttgg cttgagacga taattgcata
     1381 attgtccaaa ttgtccatca tataatgaag aattcgatgc gaaatattcc ctgtcacttt
     1441 cctttagtca acgtattctt ctccctgaag aaacagtata ctaacaatac tcacccattt
     1501 cgattttgat gttgccatac aaatagataa caagcacaat gctttcacta cgtcaatcta
     1561 taagattttt caagccagcc acaagaactt tgtgtagctc tagatatctg cttcagcaaa
     1621 aacccgtggt gaaaactgcc caaaacttag cagaagttaa tggtccagaa actttgattg
     1681 gtcctggtgc taaagagggt accgttccaa cagacctaga tcaagaaact ggtttagcta
     1741 ggttagaatt attgggtaaa ttagagggta tcgatgtttt cgacaccaaa ccattagatt
     1801 cgtccaggaa gggtaccatg aaagatccga tcatcattga atcttatgat gattatcgtt
     1861 atgtcggttg tacgggttct cccgctggtt cacatactat tatgtggtta aaaccaactg
     1921 ttaacgaagt cgccagatgc tgggaatgtg gttctgttta caaactaaac cctgttggtg
     1981 ttccaaatga tgaccaccat cacctttctc ttgaagttgc tgaagaaatg agcgaactga
     2041 tcaaagagaa catgcacatg aagctgtaca tggaaggcac cgttgacaac caccacttta
     2101 agtgcacctc tgagggtgag ggtaagccct acgaaggcac ccaaaccatg cgtatcaaag
     2161 ttgtggaggg cggtccactg ccattcgctt ttgacattct ggccaccagc ttcctgtacg
     2221 gttccaaaac cttcattaac catactcagg gcattcctga tttcttcaaa cagagctttc
     2281 cggaaggttt cacctgggag cgtgtcacca cgtatgaaga tggtggtgtg ttgaccgcca
     2341 cccaagatac ctccctgcaa gatggctgtc tgatctataa cgtgaaaatt cgtggcgtca
     2401 actttactag caatggtccc gtgatgcaga agaaaaccct gggttgggag gcctttacag
     2461 aaaccctgta tcctgccgat ggtggcctgg agggccgtaa cgacatggca ctgaagctgg
     2521 ttggtggcag ccatttgatc gcaaatgcca agaccactta cagaagcaag aaacccgcaa
     2581 aaaatctgaa gatgccgggt gtttactatg tcgactaccg tctggaaaga attaaagaag
     2641 caaataatga gacttacgtg gagcagcacg aggttgcagt cgcgagatat tgcgacttgc
     2701 ctagcaagct gggtcataaa ctgaattgag gcgcgccact tctaaataag cgaatttctt
     2761 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta
     2821 aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt
     2881 caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct ctaccggcag
     2941 atccgctagg gataacaggg taatatctgc aggtcgacaa cccttaatat aacttcgtat
     3001 aatgtatgct atacgaagtt attaggtcta gagcggccgc ggaacgatca ttcactatat
     3061 atatatcaat ttatatatac gtatgtgtaa ttgaagaaag atacgttttt tcctctattg
     3121 agaggcctgc tggatgaata gctttacctt ttctaaatcc ttgataccat cagtctctac
     3181 tcctccactt acatcgacac cgatcgcatt tggtaacata ttaatggcaa cagaaacgtt
     3241 atcaggattc aatccaccag cgataatgaa ttttatctcg ggatgacttg cagaccaact
     3301 ggaaattgca ctccaattca atttctcacc agtgccacct tcaccagaat cgaacaacgt
     3361 cagcacattg tctacgtgtt catacaggtc cagtagtaat tcacaatcct gtgggaactg
     3421 gaacctctta atgattggaa ttgaagatgg gatcaaagat ctgtattctt taatatcttc
     3481 atctccatgt aattgtatca catctagatt atattcgtgg tacagttgaa ggacatcatc
     3541 aacggactga tttctaaaca ccccgactag tttagtacct ttcacgttct cttgttggtg
     3601 aactgcagtt gaaatacctt tcgcaacaga tgagtcaatg gttcttttcc taccgggaac
     3661 acaaatgata cctaagtaat cagcaccatc atccacagca gtctttgcag cttcaacggt
     3721 ttgcaaacca cacactttaa cgagcatcac agtagatttc gcaagagggt agctcgctca
     3781 gagtacccaa gtaaatgatt agtaaactga tgtttgatag ttcaattttt caatgaaata
     3841 accttatatt aaaattgata ttactattat acaaaaataa agaataaagg atttgagttt
     3901 atacataaaa taccattatt atttgttcag tgagagatac tggggtatat gggatgtgtg
     3961 tagtgatacc atgcaatcat gtatcaaaca tgggcccggt accgagctct cgagaaccct
     4021 taatataact tcgtataatg tatgctatac gaagttatta ggtgatatca gatccactag
     4081 tggcctatgc ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa
     4141 gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4201 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4261 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4321 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4381 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4441 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4501 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4561 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4621 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4681 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4741 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4801 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4861 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4921 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4981 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5041 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5101 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5161 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5221 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5281 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5341 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5401 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5461 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5521 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5581 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5641 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5701 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5761 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5821 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5881 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5941 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6001 tactcat
//
