LOCUS       sequence-350293-        5783 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mWasabi for auxotrophic selection in the absence of 
            histidine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5783)
  TITLE     Ralser Yeast Expression plasmids for auxotrophic selection
REFERENCE   2  (bases 1 to 5783)
  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"
COMMENT     Sequence Label: pH-mWasabi
FEATURES             Location/Qualifiers
     source          1..5783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        177..843
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     713..736
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     terminator      1610..1797
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1665..1689)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     misc_feature    1864..2367
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        2759..2946
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             2947..3609
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATLPNGTNDVPSTKGVLM"
     primer_bind     complement(3675..3694)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3676..3694)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3781..3798)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3952..4540)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4032..4051)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4711..5502,5503..5571))
                     /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     5334..5353
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5572..5676)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
       61 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
      121 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgtcat
      181 tatcaatact gccatttcaa agaatacgta aataattaat agtagtgatt ttcctaactt
      241 tatttagtca aaaaattagc cttttaattc tgctgtaacc cgtacatgcc caaaataggg
      301 ggcgggttac acagaatata taacatcgta ggtgtctggg tgaacagttt attcctggca
      361 tccactaaat ataatggagc ccgcttttta agctggcatc cagaaaaaaa aagaatccca
      421 gcaccaaaat attgttttct tcaccaacca tcagttcata ggtccattct cttagcgcaa
      481 ctacagagaa caggggcaca aacaggcaaa aaacgggcac aacctcaatg gagtgatgca
      541 acctgcctgg agtaaatgat gacacaaggc aattgaccca cgcatgtatc tatctcattt
      601 tcttacacct tctattacct tctgctctct ctgatttgga aaaagctgaa aaaaaaggtt
      661 gaaaccagtt ccctgaaatt attcccctac ttgactaata agtatataaa gacggtaggt
      721 attgattgta attctgtaaa tctatttctt aaacttctta aattctactt ttatagttag
      781 tctttttttt agttttaaaa caccaagaac ttagtttcga ataaacacac ataaacaaac
      841 aaagtcgacg gatcggtgac ggtgctggtt taattaacat ggtaagtaaa ggtgaagaga
      901 caactatggg ggtaatcaaa ccagacatga agatcaaact gaaaatggaa ggtaacgtta
      961 atggccacgc gtttgttatt gaaggtgaag gagaggggaa accttacgac ggaaccaaca
     1021 ctatcaattt ggaggttaag gaaggtgcac cattaccatt cagctatgac atcttaacaa
     1081 ctgcattttc atacggcaac agagctttta ccaagtaccc tgacgatata ccaaattact
     1141 tcaaacaatc ctttcctgaa ggttattcat gggagagaac aatgacgttt gaagataagg
     1201 gcattgtcaa ggtcaaatct gacatttcta tggaagagga ttcattcatc tacgagattc
     1261 acctcaaggg cgaaaacttc ccaccaaatg gtccagtgat gcaaaaggaa actactggat
     1321 gggatgcttc tacagaaagg atgtacgtga gagatggagt cttaaaaggt gatgttaaaa
     1381 tgaagttgct acttgaaggt ggtggccatc atagagttga tttcaaaaca atctacagag
     1441 ccaaaaaggc tgtaaaactt cctgattatc attttgtgga tcatcgtata gaaattctaa
     1501 accacgacaa agattacaat aaggtcactg tttatgaaat agcagttgcc agaaattcta
     1561 cagatggaat ggatgaattg tacaaataag gcgcgccact tctaaataag cgaatttctt
     1621 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta
     1681 aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt
     1741 caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct ctaccggcag
     1801 atccgctagg gataacaggg taatatagat ctgtttagct tgcctcgtcc ccgccgggtc
     1861 accgatcgct tgcctgtaac ttacacgcgc ctcgtatctt ttaatgatgg aataatttgg
     1921 gaatttactc tgtatttatt tatttttatg ttttgtattt ggattttaga aagtaaataa
     1981 agaaggtaga agagttacgg aatgaagaaa aaaaaataaa caaaggttta aaaaatttca
     2041 acaaaaagcg tactttacat atatatttat tagacaagaa aagcagatta aatagatata
     2101 cattcgatta acgataagta aaatgtaaaa tcacaggatt ttcgtgtgtg gtcttctaca
     2161 cagacaagat gaaacaattc ggcattaata cctgagagca ggaagagcaa gataaaaggt
     2221 agtatttgtc ggcgatcccc ctagagtctt ttacatcttc ggaaaacaaa aactattttt
     2281 tctttaattc ctttttttac tttctatttt taatttatat atttatatta aaaaatttaa
     2341 attataatta tttttatagc acgtgatgaa aaggacccta agaaaccatt attatcatga
     2401 cattaaccta taaaaatagg cgcatcacga ggccctttcg tctcgcgtgt ttcggtgatg
     2461 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     2521 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcgga cgtcggggct
     2581 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataaa ttcccgtttt
     2641 aagagcttgg tgagcgctag gagtcactgc caggtatcgt ttgaacacgg cattagtcag
     2701 ggaagtcata acacagtcct ttcccgcaat tttctttttc tattactctt ggcctcctct
     2761 agtacactct atattttttt atgcctcggt aatgattttc attttttttt ttccacctag
     2821 cggatgactc tttttttttc ttagcgattg gcattatcac ataatgaatt atacattata
     2881 taaagtaatg tgatttcttc gaagaatata ctaaaaaatg agcaggcaag ataaacgaag
     2941 gcaaagatga cagagcagaa agccctagta aagcgtatta caaatgaaac caagattcag
     3001 attgcgatct ctttaaaggg tggtccccta gcgatagagc actcgatctt cccagaaaag
     3061 gaggcagaag cagtagcaga acaggccaca caatcgcaag tgattaacgt ccacacaggt
     3121 atagggtttc tggaccatat gatacatgct ctggccaagc attccggctg gtcgctaatc
     3181 gttgagtgca ttggtgactt acacatagac gaccatcaca ccactgaaga ctgcgggatt
     3241 gctctcggtc aagcttttaa agaggcccta ggggccgtgc gtggagtaaa aaggtttgga
     3301 tcaggatttg cgcctttgga tgaggcactt tccagagcgg tggtagatct ttcgaacagg
     3361 ccgtacgcag ttgtcgaact tggtttgcaa agggagaaag taggagatct ctcttgcgag
     3421 atgatcccgc attttcttga aagctttgca gaggctagca gaattaccct ccacgttgat
     3481 tgtctgcgag gcaagaatga tcatcaccgt agtgagagtg cgttcaaggc tcttgcggtt
     3541 gccataagag aagccacctt gcccaatggt accaacgatg ttccctccac caaaggtgtt
     3601 cttatgtagg agctcgaatt catcgatgat atcagatcca ctagtggcct atgcggccgc
     3661 ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca
     3721 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3781 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3841 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3901 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3961 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4021 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4081 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4141 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4201 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4261 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4321 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4381 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4441 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4501 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4561 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4621 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4681 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4741 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4801 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4861 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4921 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4981 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5041 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5101 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5161 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5221 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5281 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5341 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5401 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5461 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5521 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5581 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5641 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5701 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     5761 gccctttcgt ctcgcgcgtt tcg
//
