LOCUS       sequence-318176-        7616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of PEX3-DuDre as an additional copy in genome, use 
            auxotrophic marker TRP1(Kluyveromyces lactis). Marker for yeast 
            peroxisome..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7616)
  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 7616)
  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: ZJOM57
FEATURES             Location/Qualifiers
     source          1..7616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      3892..4079
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3947..3971)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    4131..4164
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5167..5200
                     /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(5253..5272)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5254..5272)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5359..5376)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5530..6118)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5610..5629)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6289..7080,7081..7149))
                     /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     6912..6931
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7150..7254)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7322..7340
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7378..7400)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7500..7519
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(7600..7616,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(7600..7616,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gctttgttaa tcattatcca tttttgcttt gcccttgcct
       61 tttgctttgc agcgtctaag tcgagtttgg acatgattga aaatggaggc ctaaataaca
      121 aagctaattt ctgctccttg gaaatggttt taggctttct cactaatttg gtgtactcct
      181 tgatattgcc gtcttcatca agttcgatga ccggctcttc cacatactca tattcgtcgt
      241 cgtcattttc ttcctcggag tccaaggaat ctgaagagtt gtcattgatg tatgtattag
      301 tatttgtgtc atccaatcgt tgattaaaaa tgctttcagg tcttccttcg tcaaacatgt
      361 cgttcaccct atgcaatggt tcaaatcttc cgccaatgcc gccgtaagaa attgggaatc
      421 ctgaagtttc agtaagtact tcatgtcttg cctcatcagg tggcctgaca taatcttggt
      481 caggctgttg ttgttggtaa gcttcccttt gcaacctctc cgctaattgt gcatcggcat
      541 ctgaatctac agaggtgggc gccataggag tcgaattaga cggagtaggt gtgttgttac
      601 tgctcagtaa agcagcccca ccattttcga aaaacaatga tatggctgtg ttcaagttcc
      661 cgtctgccat ttcgatgaat tgagtggcaa tagcagcgtc atcagcgcca gtgatggcca
      721 taaattcgtc tacttttccc tctgacatat aaaaggtatg ttattattgc cacacgtcgt
      781 agtctatgcg tttgtagtga cactattgag cgatcatagc taagatgtcg ttaacaagta
      841 gtagagtttg cgtgaatgta tgaatatttc cctttttcct taaaaattcc ttgcattttg
      901 gtaatcgtgt gtaatgataa gggcgttatt cgacttatat gagcatgggg tatattattg
      961 aggtagttaa tactagtcat cgtaaaagca gaagcacgaa acaaggaggc aaaccactaa
     1021 aaggatggcc ccaaatcaaa gatcacgttc gcttctgcag agacatcgag gaaaggtact
     1081 catttcattg actggaatag ctgctttatt taccacaggg tcggtggtag tgtttttcgt
     1141 gaagagatgg ttgtataaac agcagttacg gattactgag caacacttca tcaaagaaca
     1201 gatcaaaaga aggttcgagc agacacagga agactcgttg tacacaatat acgagctact
     1261 tcctgtatgg agaatggttt tgaatgaaaa cgatttgaat ttggacagca tcgttaccca
     1321 attaaaggac caaaagaacc agttgactag agcaaagtct agtgaaagca gagaaagttc
     1381 gccgttaaaa agtaaagctg agttgtggaa cgagttagaa ctaaagagtt tgatcaagct
     1441 ggtgacagtg acgtacacag tatcgtcgct gattctttta acaagactgc aactaaatat
     1501 cctgacaaga aacgagtacc tggactccgc aataaaatta accatgcagc aggaaaactg
     1561 caacaaacta cagaataggt tctataactg ggttacatcg tggtggagcg atccggagga
     1621 caaagcggat gatgcaatgg taatggcagc aaaaaagtca aagaaagaag gccaggaagt
     1681 gtatattaat gagcaagctt tcctatcact atcatggtgg attcttaaca aggggtggtt
     1741 gagttacaat gaaataatta caaatcaaat cgaaattgaa tttgacggca tacacccaag
     1801 agatactcta acgttagaag aatttagttc tcgtctcacg aacatattcc ggaacacgaa
     1861 ttcccaaata ttccagcaaa ataataataa cctcacgtct attttgcttc ctaaagattc
     1921 cagtggacag gagtttcttc tgtcacagac gcttgacgcg gatgctttaa caagttttca
     1981 ctctaacaca ctagttttta atcaattggt gaatgaactg acccaatgta tcgaaagcac
     2041 agcaacatcc atagtgttag agtctctaat aaatgaatca ttccatttca taatgaacaa
     2101 ggtcggcata aagacaatag ccaagaaaaa acctggccaa gaagatcagc aacagtatca
     2161 aatggcagtt ttcgctatgt cgatgaaaga ctgttgccaa gaaatgcttc aaacaactgc
     2221 cgggtcttcc catagcggca gcgtgaacga atacctggcc actctggatt ctgtgcagcc
     2281 gctcgatgat ctgagcgcca gcgtatacag caactttggc gtctccagct cgttttcctt
     2341 caagcctgct tcgtacgctg caggtcgtgg gattcctggg ttaattaaca tcgctagcat
     2401 ggtttccaaa ggtgaagaga acaatatggc tgtcatcaag cccttcatga gattcaaggt
     2461 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggcaa
     2521 gccctacgag ggcacccaga ccgccaagct gcaggtgacc aagggcggcc ccctgccctt
     2581 cgcctgggac atcctgtccc cccagttcca gtacggctcc aaggtgtacg tgaagcaccc
     2641 cgccgacatc cccgactaca agaagctgtc cttccccgag ggcttcaagt gggagagagt
     2701 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     2761 caccttcatc taccacgtga agttcatcgg cgtgaacttc ccctccgacg gccccgtaat
     2821 gcagaagaag actctgggct gggagccctc caccgagaga ctgtacccca gagacggcgt
     2881 gctgaagggc gagatccaca aggcgctgaa gctgaagggc ggcggccact acctggtgga
     2941 gttcaagtca atctacatgg ccaagaagcc cgtgaagctg cccggctact actacgtgga
     3001 ctccaagctg gacatcacct cccacaacga ggactacacc gtggtggagc agtacgagag
     3061 agccgaggcc agacaccacc tgttcttaat tccagttcca acgcgtgatc ttgagaagcc
     3121 attcaacgtc atcaagccct tcatgagatt caaggtgcac atggagggct ccgtgaacgg
     3181 ccacgagttc gagatcgagg gcgagggcga gggcaagccc tacgagggca cccagaccgc
     3241 caagctgcag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca
     3301 gttccagtac ggctccaagg tgtacgtgaa gcaccccgcc gacatccccg actacaagaa
     3361 gctgtccttc cccgagggct tcaagtggga gagagtgatg aacttcgagg acggcggcgt
     3421 ggtgaccgtg acccaggact cctccctgca ggacggcacc ttcatctacc acgtgaagtt
     3481 catcggcgtg aacttcccct ccgacggccc cgtaatgcag aagaagactc tgggctggga
     3541 gccctccacc gagagactgt accccagaga cggcgtgctg aagggcgaga tccacaaggc
     3601 gctgaagctg aagggcggcg gccactacct ggtggagttc aagtcaatct acatggccaa
     3661 gaagcccgtg aagctgcccg gctactacta cgtggactcc aagctggaca tcacctccca
     3721 caacgaggac tacaccgtgg tggagcagta cgagagagcc gaggccagac accacctgtt
     3781 cttaattgtc gttctgaaca gagctgagac tccattgcca ttggatccaa ccggtaaggt
     3841 caaggctgag ctggatacta ggcctaggtg aggcgcgcca cttctaaata agcgaatttc
     3901 ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt
     3961 taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag
     4021 gtcaggttgc tttctcaggt atagtatgag gtcgctctta ttgaccacac ctctaccggc
     4081 agatccgcta gggataacag ggtaatatct gcaggtcgac aacccttaat ataacttcgt
     4141 ataatgtatg ctatacgaag ttattaggtc tagagcggcc gcggaacgat cattcactat
     4201 atatatatca atttatatat acgtatgtgt aattgaagaa agatacgttt tttcctctat
     4261 tgagaggcct gctggatgaa tagctttacc ttttctaaat ccttgatacc atcagtctct
     4321 actcctccac ttacatcgac accgatcgca tttggtaaca tattaatggc aacagaaacg
     4381 ttatcaggat tcaatccacc agcgataatg aattttatct cgggatgact tgcagaccaa
     4441 ctggaaattg cactccaatt caatttctca ccagtgccac cttcaccaga atcgaacaac
     4501 gtcagcacat tgtctacgtg ttcatacagg tccagtagta attcacaatc ctgtgggaac
     4561 tggaacctct taatgattgg aattgaagat gggatcaaag atctgtattc tttaatatct
     4621 tcatctccat gtaattgtat cacatctaga ttatattcgt ggtacagttg aaggacatca
     4681 tcaacggact gatttctaaa caccccgact agtttagtac ctttcacgtt ctcttgttgg
     4741 tgaactgcag ttgaaatacc tttcgcaaca gatgagtcaa tggttctttt cctaccggga
     4801 acacaaatga tacctaagta atcagcacca tcatccacag cagtctttgc agcttcaacg
     4861 gtttgcaaac cacacacttt aacgagcatc acagtagatt tcgcaagagg gtagctcgct
     4921 cagagtaccc aagtaaatga ttagtaaact gatgtttgat agttcaattt ttcaatgaaa
     4981 taaccttata ttaaaattga tattactatt atacaaaaat aaagaataaa ggatttgagt
     5041 ttatacataa aataccatta ttatttgttc agtgagagat actggggtat atgggatgtg
     5101 tgtagtgata ccatgcaatc atgtatcaaa catgggcccg gtaccgagct ctcgagaacc
     5161 cttaatataa cttcgtataa tgtatgctat acgaagttat taggtgatat cagatccact
     5221 agtggcctat gcggccgcgg atctgccggt ctccctatag tgagtcgtat taatttcgat
     5281 aagccaggtt aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5341 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5401 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5461 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5521 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5581 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5641 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5701 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5761 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5821 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5881 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5941 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6001 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6061 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6121 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6181 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6241 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6301 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6361 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6421 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6481 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6541 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6601 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6661 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6721 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6781 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6841 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6901 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6961 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7021 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7081 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7141 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7201 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7261 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     7321 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     7381 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     7441 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     7501 ggcatcagag cagattgtac tgagagtgca ccatatggac atattgtcgt tagaacgcgg
     7561 ctacaattaa tacataacct tatgtatcat acacatacga tttaggtgac actata
//
