LOCUS       sequence-256885-        5722 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 5722)
  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 5722)
  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..5722
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1995..2182
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2050..2074)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    2234..2267
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3273..3306
                     /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(3359..3378)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3360..3378)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3465..3482)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3636..4224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3716..3735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4395..5186,5187..5255))
                     /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     5018..5037
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5256..5360)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5428..5446
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5484..5506)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5606..5625
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(5706..5722,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(5706..5722,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg tgggattcct gggttaatta
       61 acatcgctag cacactttca aagtttgtgg ttgagcagtc gtagggtcgg atggcttgta
      121 gaccagttcc aaagatcctg ggcctgggat cagtgtgtcc gtagctttat attgatcacc
      181 gtgggcgtgt cttccaataa tgattggttt ttcccaacgt gggaccagtc taggaattct
      241 aggaatcaca atgggctctc tgaacactgt accgccgaga atgtttctga tggtaccatt
      301 aggagatttc cacatcttgt gcaggttgaa ttccttcaca cgagcttcat caggagtgat
      361 ggtggcacat ttgataccaa caccatactt cttgatcgcc tcagcagcat cctgagtaat
      421 cttgtcggag gtggcgtcac gagattcgac agataagtcg tagtacttca aatctacgtc
      481 caagtagggt agaatcaatt tcttcttgat cttatcccaa atgatacggg tcatttcatc
      541 accgtccaac tcgacaacgg gttgtttgac cttaatctta ctgaaagcag caaggcgaga
      601 ggtggaaaat aatcttctag ataacatact catgctgggc gatcttcctt gtgcgtctgt
      661 tgtctcacaa ttgcttgaag gaagatttca taagatcata tgagtccctc tttatatggg
      721 caagtggaat tatgtgagtc aaaatccgcg cgtgacccgt aaagcgttat cagaaggtgc
      781 aaacggtgct atttagctca taaaaggaat gattcaagct cttttggatt gtaagacacc
      841 tttatttagt ccaagatcat tgcagaccct tgttatggtc tttagcagag tcctcctcta
      901 tatctcttca tttactgcaa cctgattggc ccgctaccac gatgcccgct ttgttcctgt
      961 ggtattaaaa gaatcgatga aagagactct tatcttcagg gaaaattagg accgagaatt
     1021 agagcaagca agatatttgc aaactactaa ctacaagcga cttacacaga catgactatt
     1081 gctccaatta ctggtacgat caagagaaga gtcatcatgg acatcgtcct cgggttctcc
     1141 ctcgggggtg tcatggcctc ttactggtgg tggggattcc acatggataa gattaacaag
     1201 agagagaagt tctacgcaga gctagctgag aggaaaaagc aagagaacct ttctcttgaa
     1261 gttgctgaag aaatgagcga actgatcaaa gagaacatgc acatgaagct gtacatggaa
     1321 ggcaccgttg acaaccacca ctttaagtgc acctctgagg gtgagggtaa gccctacgaa
     1381 ggcacccaaa ccatgcgtat caaagttgtg gagggcggtc cactgccatt cgcttttgac
     1441 attctggcca ccagcttcct gtacggttcc aaaaccttca ttaaccatac tcagggcatt
     1501 cctgatttct tcaaacagag ctttccggaa ggtttcacct gggagcgtgt caccacgtat
     1561 gaagatggtg gtgtgttgac cgccacccaa gatacctccc tgcaagatgg ctgtctgatc
     1621 tataacgtga aaattcgtgg cgtcaacttt actagcaatg gtcccgtgat gcagaagaaa
     1681 accctgggtt gggaggcctt tacagaaacc ctgtatcctg ccgatggtgg cctggagggc
     1741 cgtaacgaca tggcactgaa gctggttggt ggcagccatt tgatcgcaaa tgccaagacc
     1801 acttacagaa gcaagaaacc cgcaaaaaat ctgaagatgc cgggtgttta ctatgtcgac
     1861 taccgtctgg aaagaattaa agaagcaaat aatgagactt acgtggagca gcacgaggtt
     1921 gcagtcgcga gatattgcga cttgcctagc aagctgggtc ataaactgaa ttgaggcgcg
     1981 ccacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
     2041 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
     2101 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat gaggtcgctc
     2161 ttattgacca cacctctacc ggcagatccg ctagggataa cagggtaata tctgcaggtc
     2221 gacaaccctt aatataactt cgtataatgt atgctatacg aagttattag gtctagagcg
     2281 gccgcggaac gatcattcac tatatatata tcaatttata tatacgtatg tgtaattgaa
     2341 gaaagatacg ttttttcctc tattgagagg cctgctggat gaatagcttt accttttcta
     2401 aatccttgat accatcagtc tctactcctc cacttacatc gacaccgatc gcatttggta
     2461 acatattaat ggcaacagaa acgttatcag gattcaatcc accagcgata atgaatttta
     2521 tctcgggatg acttgcagac caactggaaa ttgcactcca attcaatttc tcaccagtgc
     2581 caccttcacc agaatcgaac aacgtcagca cattgtctac gtgttcatac aggtccagta
     2641 gtaattcaca atcctgtggg aactggaacc tcttaatgat tggaattgaa gatgggatca
     2701 aagatctgta ttctttaata tcttcatctc catgtaattg tatcacatct agattatatt
     2761 cgtggtacag ttgaaggaca tcatcaacgg actgatttct aaacaccccg actagtttag
     2821 tacctttcac gttctcttgt tggtgaactg cagttgaaat acctttcgca acagatgagt
     2881 caatggttct tttcctaccg ggaacacaaa tgatacctaa gtaatcagca ccatcatcca
     2941 cagcagtctt tgcagcttca acggtttgca aaccacacac tttaacgagc atcacagtag
     3001 atttcgcaag agggtagctc gctcagagta cccaagtaaa tgattagtaa actgatgttt
     3061 gatagttcaa tttttcaatg aaataacctt atattaaaat tgatattact attatacaaa
     3121 aataaagaat aaaggatttg agtttataca taaaatacca ttattatttg ttcagtgaga
     3181 gataccgggg tatatgggat gtgtgtagtg ataccatgca atcatgtatc aaacatgggc
     3241 ccggtaccga gctctggtcg agaaccctta atataacttc gtataatgta tgctatacga
     3301 agttattagg tgatatcaga tccactagtg gcctatgcgg ccgcggatct gccggtctcc
     3361 ctatagtgag tcgtattaat ttcgataagc caggttaacc tgcattaatg aatcggccaa
     3421 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3481 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3541 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3601 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3661 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3721 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3781 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc
     3841 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3901 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3961 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4021 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     4081 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4141 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4201 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4261 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4321 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4381 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4441 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4501 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4561 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4621 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4681 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4741 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4801 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4861 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4921 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4981 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5041 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5101 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5161 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5221 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5281 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5341 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     5401 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     5461 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     5521 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     5581 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     5641 atggacatat tgtcgttaga acgcggctac aattaataca taaccttatg tatcatacac
     5701 atacgattta ggtgacacta ta
//
