LOCUS       sequence-350305-        5704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mWasabi for auxotrophic selection in the absence of 
            uracil.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5704)
  TITLE     Ralser Yeast Expression plasmids for auxotrophic selection
REFERENCE   2  (bases 1 to 5704)
  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: pU-mWasabi
FEATURES             Location/Qualifiers
     source          1..5704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     139..158
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     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"
     CDS             2727..3530
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     primer_bind     complement(3596..3615)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3597..3615)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3702..3719)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3873..4461)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3953..3972)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4632..5423,5424..5492))
                     /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     5255..5274
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5493..5597)
                     /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 ctcgcatctt 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 agtatttgtt ggcgatcccc ctagagtctt ttacatcttc ggaaaacaaa aactattttt
     2281 tctttaattt ctttttttac tttctatttt taatttatat atttatatta aaaaatttaa
     2341 attataatta tttttatagc acgtgatgaa aaggacccta agaaaccatt attatcatga
     2401 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     2461 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     2521 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggc gattcggtaa
     2581 tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca
     2641 tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa
     2701 aaacctgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta
     2761 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact
     2821 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag
     2881 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg
     2941 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaagaca
     3001 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa
     3061 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg
     3121 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag
     3181 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg
     3241 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag
     3301 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagacg
     3361 cattgggtca acagtataga accgtggatg atgtggtctc tacaggatct gacattatta
     3421 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca
     3481 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa gagctcgaat
     3541 tcatcgatga tatcagatcc actagtggcc tatgcggccg cggatctgcc ggtctcccta
     3601 tagtgagtcg tattaatttc gataagccag gttaacctgc attaatgaat cggccaacgc
     3661 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3721 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3781 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3841 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3901 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3961 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4021 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4081 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4141 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4201 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4261 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4321 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4381 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4441 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4501 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4561 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4621 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4681 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4741 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4801 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4861 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4921 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4981 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5041 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5101 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5161 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5221 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5281 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5341 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5401 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5461 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5521 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5581 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     5641 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     5701 ttcg
//
