LOCUS       sequence-265880-        5719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of COX9-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 5719)
  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 5719)
  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: ZJOM141
FEATURES             Location/Qualifiers
     source          1..5719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(73..106)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(1109..1142)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      complement(1194..1381)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     1302..1326
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        complement(3374..3392)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3375..3392)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3473..3492)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3592..3614
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3652..3670)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3738..3842
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3843..3911,3912..4703)
                     /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     complement(4061..4080)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4874..5462
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5363..5382
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5616..5633
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatacgact cactataggg agaccggcag atccgcggcc gcataggcca ctagtggatc
       61 tgatatcacc taataacttc gtatagcata cattatacga agttatatta agggttctcg
      121 agagctcggt accgggccca tgtttgatac atgattgcat ggtatcacta cacacatccc
      181 atatacccca gtatctctca ctgaacaaat aataatggta ttttatgtat aaactcaaat
      241 cctttattct ttatttttgt ataatagtaa tatcaatttt aatataaggt tatttcattg
      301 aaaaattgaa ctatcaaaca tcagtttact aatcatttac ttgggtactc tgagcgagct
      361 accctcttgc gaaatctact gtgatgctcg ttaaagtgtg tggtttgcaa accgttgaag
      421 ctgcaaagac tgctgtggat gatggtgctg attacttagg tatcatttgt gttcccggta
      481 ggaaaagaac cattgactca tctgttgcga aaggtatttc aactgcagtt caccaacaag
      541 agaacgtgaa aggtactaaa ctagtcgggg tgtttagaaa tcagtccgtt gatgatgtcc
      601 ttcaactgta ccacgaatat aatctagatg tgatacaatt acatggagat gaagatatta
      661 aagaatacag atctttgatc ccatcttcaa ttccaatcat taagaggttc cagttcccac
      721 aggattgtga attactactg gacctgtatg aacacgtaga caatgtgctg acgttgttcg
      781 attctggtga aggtggcact ggtgagaaat tgaattggag tgcaatttcc agttggtctg
      841 caagtcatcc cgagataaaa ttcattatcg ctggtggatt gaatcctgat aacgtttctg
      901 ttgccattaa tatgttacca aatgcgatcg gtgtcgatgt aagtggagga gtagagactg
      961 atggtatcaa ggatttagaa aaggtaaagc tattcatcca gcaggcctct caatagagga
     1021 aaaaacgtat ctttcttcaa ttacacatac gtatatataa attgatatat atatagtgaa
     1081 tgatcgttcc gcggccgctc tagacctaat aacttcgtat agcatacatt atacgaagtt
     1141 atattaaggg ttgtcgacct gcagatatta ccctgttatc cctagcggat ctgccggtag
     1201 aggtgtggtc aataagagcg acctcatact atacctgaga aagcaacctg acctacagga
     1261 aagagttact caagaataag aattttcgtt ttaaaaccta agagtcactt taaaatttgt
     1321 atacacttat tttttttata acttatttaa taataaaaat cataaatcat aagaaattcg
     1381 cttatttaga agtggcgcgc ctcaattcag tttatgaccc agcttgctag gcaagtcgca
     1441 atatctcgcg actgcaacct cgtgctgctc cacgtaagtc tcattatttg cttctttaat
     1501 tctttccaga cggtagtcga catagtaaac acccggcatc ttcagatttt ttgcgggttt
     1561 cttgcttctg taagtggtct tggcatttgc gatcaaatgg ctgccaccaa ccagcttcag
     1621 tgccatgtcg ttacggccct ccaggccacc atcggcagga tacagggttt ctgtaaaggc
     1681 ctcccaaccc agggttttct tctgcatcac gggaccattg ctagtaaagt tgacgccacg
     1741 aattttcacg ttatagatca gacagccatc ttgcagggag gtatcttggg tggcggtcaa
     1801 cacaccacca tcttcatacg tggtgacacg ctcccaggtg aaaccttccg gaaagctctg
     1861 tttgaagaaa tcaggaatgc cctgagtatg gttaatgaag gttttggaac cgtacaggaa
     1921 gctggtggcc agaatgtcaa aagcgaatgg cagtggaccg ccctccacaa ctttgatacg
     1981 catggtttgg gtgccttcgt agggcttacc ctcaccctca gaggtgcact taaagtggtg
     2041 gttgtcaacg gtgccttcca tgtacagctt catgtgcatg ttctctttga tcagttcgct
     2101 catttcttca gcaacttcaa gagaaaggtt ctcttgcttt ttcctctcag ctagctctgc
     2161 gtagaacttc tctctcttgt taatcttatc catgtggaat ccccaccacc agtaagaggc
     2221 catgacaccc ccgagggaga acccgaggac gatgtccatg atgactcttc tcttgatcgt
     2281 accagtaatt ggagcaatag tcatgtctgt gtaagtcgct tgtagttagt agtttgcaaa
     2341 tatcttgctt gctctaattc tcggtcctaa ttttccctga agataagagt ctctttcatc
     2401 gattctttta ataccacagg aacaaagcgg gcatcgtggt agcgggccaa tcaggttgca
     2461 gtaaatgaag agatatagag gaggactctg ctaaagacca taacaagggt ctgcaatgat
     2521 cttggactaa ataaaggtgt cttacaatcc aaaagagctt gaatcattcc ttttatgagc
     2581 taaatagcac cgtttgcacc ttctgataac gctttacggg tcacgcgcgg attttgactc
     2641 acataattcc acttgcccat ataaagaggg actcatatga tcttatgaaa tcttccttca
     2701 agcaattgtg agacaacaga cgcacaagga agatcgccca gcatgagtat gttatctaga
     2761 agattatttt ccacctctcg ccttgctgct ttcagtaaga ttaaggtcaa acaacccgtt
     2821 gtcgagttgg acggtgatga aatgacccgt atcatttggg ataagatcaa gaagaaattg
     2881 attctaccct acttggacgt agatttgaag tactacgact tatctgtcga atctcgtgac
     2941 gccacctccg acaagattac tcaggatgct gctgaggcga tcaagaagta tggtgttggt
     3001 atcaaatgtg ccaccatcac tcctgatgaa gctcgtgtga aggaattcaa cctgcacaag
     3061 atgtggaaat ctcctaatgg taccatcaga aacattctcg gcggtacagt gttcagagag
     3121 cccattgtga ttcctagaat tcctagactg gtcccacgtt gggaaaaacc aatcattatt
     3181 ggaagacacg cccacggtga tcaatataaa gctacggaca cactgatccc aggcccagga
     3241 tctttggaac tggtctacaa gccatccgac cctacgactg ctcaaccaca aactttgaaa
     3301 gtgtgctagc gatgttaatt aacccaggaa tcccacgacc tgcagcgtac gaagcttcag
     3361 ctggcggccg cgttctatag tgtcacctaa atcgtatgtg tatgatacat aaggttatgt
     3421 attaattgta gccgcgttct aacgacaata tgtccatatg gtgcactctc agtacaatct
     3481 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     3541 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3601 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     3661 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     3721 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3781 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3841 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3901 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3961 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4021 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4081 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4141 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4201 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4261 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4321 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4381 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4441 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4501 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4561 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4621 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4681 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4741 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4801 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4861 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4921 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4981 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5041 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5101 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5161 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5221 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5281 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5341 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5401 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5461 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5521 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     5581 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     5641 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcaggt
     5701 taacctggct tatcgaaat
//
