LOCUS       sequence-256884-        6010 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 6010)
  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 6010)
  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..6010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1072..1137
                     /codon_start=1
                     /product="mitochondrial presequence of yeast cytochrome c 
                     oxidase subunit IV (Hurt et al., 1984)"
                     /label=pre-CoxIV
                     /translation="MLSLRQSIRFFKPATRTLCSSR"
     terminator      2283..2470
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2338..2362)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    2522..2555
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3561..3594
                     /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(3647..3666)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3648..3666)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3753..3770)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3924..4512)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4004..4023)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4683..5474,5475..5543))
                     /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     5306..5325
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5544..5648)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5716..5734
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5772..5794)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5894..5913
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(5994..6010,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(5994..6010,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg tgggattcct gggttaatta
       61 acatcgctag cgcctattgg attggcccaa ttgctgccaa atttggtgaa tatggtggtg
      121 atgtcgcaat gtggttaagt atggcgttct caggtgttgt ataccctcct tgtagatact
      181 tagagctacg taaatttggt cgttaattga aatgacataa gtatatatta aaactacggt
      241 gcagcggcaa actttagttt agataatatt ggcaggtaga ttcatggtat gttaaattac
      301 gctcagggag gatatttttt tgtacacatt atcttttcac atgaaaatct aagcctggtg
      361 gacaattgaa atcatcgagt gatatactga cactttgaac agtagcacaa atcgtttaat
      421 ttgctgtcac gatctaagtt ttctgtcagc tgactgtcca ccaataagat cagcggattc
      481 caaaggcgtc cttcgaaaaa gcgatatctc ccatcttggt tagtagcaaa taaagggaag
      541 agaaaataag acacactcaa ggataaaata cttcgcttct tccaagtata aacacaggaa
      601 agaaagatat acgaagatta gaattttttt catacttttt acaaaacatt tttttgtact
      661 taggcacgct aattgcgtaa aaattagcat aaacaggaaa taagattatt ttgaaatgta
      721 tgttataatc ctcttattga aaaatacatg agatgtagga gaagaactac caggaaagaa
      781 gatatttaac agatcgaacg aaacggaaaa gaactaagag aaaacaaatc tggatttgac
      841 tttaagaaaa aatttatttg caagtttaac tcgttccctg cttttctatt tggcttgaga
      901 cgataattgc ataattgtcc aaattgtcca tcatataatg aagaattcga tgcgaaatat
      961 tccctgtcac tttcctttag tcaacgtatt cttctccctg aagaaacagt atactaacaa
     1021 tactcaccca tttcgatttt gatgttgcca tacaaataga taacaagcac aatgctttca
     1081 ctacgtcaat ctataagatt tttcaagcca gccacaagaa ctttgtgtag ctctagatat
     1141 ctgcttcagc aaaaacccgt ggtgaaaact gcccaaaact tagcagaagt taatggtcca
     1201 gaaactttga ttggtcctgg tgctaaagag ggtaccgttc caacagacct agatcaagaa
     1261 actggtttag ctaggttaga attattgggt aaattagagg gtatcgatgt tttcgacacc
     1321 aaaccattag attcgtccag gaagggtacc atgaaagatc cgatcatcat tgaatcttat
     1381 gatgattatc gttatgtcgg ttgtacgggt tctcccgctg gttcacatac tattatgtgg
     1441 ttaaaaccaa ctgttaacga agtcgccaga tgctgggaat gtggttctgt ttacaaacta
     1501 aaccctgttg gtgttccaaa tgatgaccac catcaccttt ctcttgaagt tgctgaagaa
     1561 atgagcgaac tgatcaaaga gaacatgcac atgaagctgt acatggaagg caccgttgac
     1621 aaccaccact ttaagtgcac ctctgagggt gagggtaagc cctacgaagg cacccaaacc
     1681 atgcgtatca aagttgtgga gggcggtcca ctgccattcg cttttgacat tctggccacc
     1741 agcttcctgt acggttccaa aaccttcatt aaccatactc agggcattcc tgatttcttc
     1801 aaacagagct ttccggaagg tttcacctgg gagcgtgtca ccacgtatga agatggtggt
     1861 gtgttgaccg ccacccaaga tacctccctg caagatggct gtctgatcta taacgtgaaa
     1921 attcgtggcg tcaactttac tagcaatggt cccgtgatgc agaagaaaac cctgggttgg
     1981 gaggccttta cagaaaccct gtatcctgcc gatggtggcc tggagggccg taacgacatg
     2041 gcactgaagc tggttggtgg cagccatttg atcgcaaatg ccaagaccac ttacagaagc
     2101 aagaaacccg caaaaaatct gaagatgccg ggtgtttact atgtcgacta ccgtctggaa
     2161 agaattaaag aagcaaataa tgagacttac gtggagcagc acgaggttgc agtcgcgaga
     2221 tattgcgact tgcctagcaa gctgggtcat aaactgaatt gaggcgcgcc acttctaaat
     2281 aagcgaattt cttatgattt atgattttta ttattaaata agttataaaa aaaataagtg
     2341 tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa attcttattc ttgagtaact
     2401 ctttcctgta ggtcaggttg ctttctcagg tatagtatga ggtcgctctt attgaccaca
     2461 cctctaccgg cagatccgct agggataaca gggtaatatc tgcaggtcga caacccttaa
     2521 tataacttcg tataatgtat gctatacgaa gttattaggt ctagagcggc cgcggaacga
     2581 tcattcacta tatatatatc aatttatata tacgtatgtg taattgaaga aagatacgtt
     2641 ttttcctcta ttgagaggcc tgctggatga atagctttac cttttctaaa tccttgatac
     2701 catcagtctc tactcctcca cttacatcga caccgatcgc atttggtaac atattaatgg
     2761 caacagaaac gttatcagga ttcaatccac cagcgataat gaattttatc tcgggatgac
     2821 ttgcagacca actggaaatt gcactccaat tcaatttctc accagtgcca ccttcaccag
     2881 aatcgaacaa cgtcagcaca ttgtctacgt gttcatacag gtccagtagt aattcacaat
     2941 cctgtgggaa ctggaacctc ttaatgattg gaattgaaga tgggatcaaa gatctgtatt
     3001 ctttaatatc ttcatctcca tgtaattgta tcacatctag attatattcg tggtacagtt
     3061 gaaggacatc atcaacggac tgatttctaa acaccccgac tagtttagta cctttcacgt
     3121 tctcttgttg gtgaactgca gttgaaatac ctttcgcaac agatgagtca atggttcttt
     3181 tcctaccggg aacacaaatg atacctaagt aatcagcacc atcatccaca gcagtctttg
     3241 cagcttcaac ggtttgcaaa ccacacactt taacgagcat cacagtagat ttcgcaagag
     3301 ggtagctcgc tcagagtacc caagtaaatg attagtaaac tgatgtttga tagttcaatt
     3361 tttcaatgaa ataaccttat attaaaattg atattactat tatacaaaaa taaagaataa
     3421 aggatttgag tttatacata aaataccatt attatttgtt cagtgagaga taccggggta
     3481 tatgggatgt gtgtagtgat accatgcaat catgtatcaa acatgggccc ggtaccgagc
     3541 tctggtcgag aacccttaat ataacttcgt ataatgtatg ctatacgaag ttattaggtg
     3601 atatcagatc cactagtggc ctatgcggcc gcggatctgc cggtctccct atagtgagtc
     3661 gtattaattt cgataagcca ggttaacctg cattaatgaa tcggccaacg cgcggggaga
     3721 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3781 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3841 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3901 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3961 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4021 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4081 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     4141 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4201 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4261 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4321 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     4381 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4441 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4501 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4561 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4621 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4681 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4741 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4801 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4861 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4921 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4981 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5041 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5101 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5161 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5221 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5281 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5341 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5401 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5461 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5521 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5581 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5641 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     5701 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     5761 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     5821 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     5881 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat ggacatattg
     5941 tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg
     6001 tgacactata
//
