LOCUS       sequence-350306-        6683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of YLL058W from Zygosaccharomyces rouxii for auxotrophic 
            selection in the absence of uracil.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6683)
  TITLE     Ralser Yeast Expression plasmids for auxotrophic selection
REFERENCE   2  (bases 1 to 6683)
  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-YLL058W-zr
FEATURES             Location/Qualifiers
     source          1..6683
                     /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      2589..2776
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2644..2668)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     misc_feature    2843..3346
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             3706..4509
                     /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(4575..4594)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4576..4594)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4681..4698)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4852..5440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4932..4951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5611..6402,6403..6471))
                     /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     6234..6253
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6472..6576)
                     /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 aaagtcgaca tgactgaaat cgcattcggc cagccactgc cgtctaatct ggaatacgct
      901 gtttccttcg gtattccgac atgggactct gcgattgggt atgccgaaaa agaccccaga
      961 gtggtatcta aaatggcaac aggatacccg aggtattttc cccaacctcc tgtacaaaaa
     1021 ttgtgtaagc acttccttaa aaagtttggc agagaaagtg aggattgtag gccattcccg
     1081 tcactgaagg aagcccagga gtgtttgaaa ttcgtcaagt atgtgtccgg gtcagagacc
     1141 ttggtgcata tcgaggttga ggcctttgag tttgaaaaca atgaaggtaa cagttcagaa
     1201 agttctgata aaaacgtaac cattgcggcg gtattggcct cagggaatga gttcgatata
     1261 gtaagagagt actggaagct aagaggagag tgcgtttctt cccgtctggc cacgaatatc
     1321 aaccaacttc ttgataacat gtccacaagt tcaaatgaaa aggaacatga agcggaggac
     1381 ggtatattgt ttgctagaaa agagggcgag aacgctaaaa tacttttaaa gaaaaggatt
     1441 gttgaaaacc actgttcccc gtttggttta ccccgtaaaa atagtgtttg ggacaaggag
     1501 agtatcgatc ccgacaaaga cgtatatttg gtgtcatcag gcatgtctag tatattcgca
     1561 gcacgtcgtt ttttgacttt ttgggaggag aagcgtgcat ccgactgtag ctcaacccaa
     1621 gacaatttta agggaaaaaa agatgtacta ctttgcaaca ctgccgtggt gttcggattc
     1681 ccatttaagg acacacaggt gatcatgaaa acttttggga aatgccaatt ctacgggttc
     1741 ggcgacagta gggacatctc ttctttgaag aagtttctgg actctaataa aaggatatta
     1801 gccgtattta ttgaaactcc atccaacccc ctgttgaaca tgccagatct gcctcagcta
     1861 cgtaagcttg cagacgaata tggttttttt ataataattg atgatactat aggaggtctt
     1921 aacgtagata cactgtccta tgctgacatc cttagcacgt ctctaacgaa gctgttcaat
     1981 gggtcttcta acgtgatggg gggatccatt attctaaatc cgaagtcatt tttttatcca
     2041 tgcgcaaggg agtatttcgc ctcttctaaa tatgaagacc tgctgtgggg agaggatgcc
     2101 gtagtacttg aggagaattc ccgtgatttt gaagaaagga caatttgtgc caacaaaaat
     2161 acggagacat tgcttaacga gcttctgcta ccgctagagg ggaacgaatt caagaagatt
     2221 tattacccaa gtgtgagcag caaagagaca ttcgcgaact atgagacggt taggaacaaa
     2281 aaaggggggt acggatgcct attcagcatg agtttcttcg acgaaaaaga cgccttgatc
     2341 ttctacgatt cccttaaggt tttcaaaggg ccatctaacg gcactaattt cacattagct
     2401 tgtccatatg tccacctggc gcaccactcc gaactagaaa aagtcagtga gtttggggta
     2461 gacccgaact ttattcgtgt ttcagtgggt cttgaaaata ttaaatggtt aatagacgtg
     2521 ttttccaacg ccatcgaggc ggtcagaaaa aggagactac agacatgagg cgcgccactt
     2581 ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa
     2641 taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga
     2701 gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc gctcttattg
     2761 accacacctc taccggcaga tccgctaggg ataacagggt aatatagatc tgtttagctt
     2821 gcctcgtccc cgccgggtca ccgatcgctt gcctgtaact tacacgcgcc tcgcatcttt
     2881 taatgatgga ataatttggg aatttactct gtatttattt atttttatgt tttgtatttg
     2941 gattttagaa agtaaataaa gaaggtagaa gagttacgga atgaagaaaa aaaaataaac
     3001 aaaggtttaa aaaatttcaa caaaaagcgt actttacata tatatttatt agacaagaaa
     3061 agcagattaa atagatatac attcgattaa cgataagtaa aatgtaaaat cacaggattt
     3121 tcgtgtgtgg tcttctacac agacaagatg aaacaattcg gcattaatac ctgagagcag
     3181 gaagagcaag ataaaaggta gtatttgttg gcgatccccc tagagtcttt tacatcttcg
     3241 gaaaacaaaa actatttttt ctttaatttc tttttttact ttctattttt aatttatata
     3301 tttatattaa aaaatttaaa ttataattat ttttatagca cgtgatgaaa aggaccctaa
     3361 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     3421 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     3481 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     3541 gttggcggcg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
     3601 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
     3661 acccaactgc acagaacaaa aacctgcagg aaacgaagat aaatcatgtc gaaagctaca
     3721 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg
     3781 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg
     3841 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg
     3901 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat
     3961 tttttactct tcgaagacag aaaatttgct gacattggta atacagtcaa attgcagtac
     4021 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg
     4081 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga
     4141 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact
     4201 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa
     4261 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt
     4321 ttagatgaca agggagacgc attgggtcaa cagtatagaa ccgtggatga tgtggtctct
     4381 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag
     4441 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag
     4501 caaaactaag agctcgaatt catcgatgat atcagatcca ctagtggcct atgcggccgc
     4561 ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca
     4621 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4681 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4741 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4801 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4861 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4921 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4981 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5041 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5101 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5161 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5221 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5281 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5341 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5401 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5461 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5521 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5581 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5641 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5701 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5761 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5821 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5881 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5941 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6001 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6061 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6121 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6181 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6241 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6301 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6361 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6421 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6481 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6541 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6601 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     6661 gccctttcgt ctcgcgcgtt tcg
//
