LOCUS       sequence-350316-        5695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of TagRFP657 for auxotrophic selection in the absence of 
            uracil.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5695)
  TITLE     Ralser Yeast Expression plasmids for auxotrophic selection
REFERENCE   2  (bases 1 to 5695)
  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-TagRFP657
FEATURES             Location/Qualifiers
     source          1..5695
                     /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      1601..1788
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1656..1680)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     misc_feature    1855..2358
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             2718..3521
                     /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(3587..3606)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3588..3606)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3693..3710)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3864..4452)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3944..3963)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4623..5414,5415..5483))
                     /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     5246..5265
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5484..5588)
                     /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 gagcgaattg atcacagaaa
      901 atatgcacat gaagttgtat atggagggaa ccgtcaataa ccatcatttc aaatgcactt
      961 ctgaaggtga gggcaaacct tacgaaggga cccaaacaca gagaattaag gtagtagaag
     1021 ggggtccact gccatttgcc tttgacatcc tagcaacatc cttcatgtac ggttcccaca
     1081 cattcataaa ccacactcaa ggaataccag acttttggaa gcagagtttt ccagaaggtt
     1141 ttacgtggga aagagttact acttacgagg acggtggtgt attaacagcc acacaagata
     1201 catcattaca agatggttgt ctgatctaca acgttaaaat cagaggtgtt aactttcctt
     1261 caaatgggcc agttatgcaa aagaaaactt taggctggga ggcccatact gaaatgcttt
     1321 acccagcaga tggcggcttg gaaggcagga cggctttggc tttaaagcta gttggtggag
     1381 gacatttgat ttgtaatttc aaaacaactt acagaagcaa aaagcctgct aaaaatctta
     1441 agatgccagg agtgtactat gtcgattaca gattggaacg tattaaagag gctgataagg
     1501 aaacctatgt ggaacaacat gaagtggcag tcgcgagata ttgtgatctt ccttctaaac
     1561 taggtcataa actcaattaa ggcgcgccac ttctaaataa gcgaatttct tatgatttat
     1621 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     1681 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
     1741 ttctcaggta tagtatgagg tcgctcttat tgaccacacc tctaccggca gatccgctag
     1801 ggataacagg gtaatataga tctgtttagc ttgcctcgtc cccgccgggt caccgatcgc
     1861 ttgcctgtaa cttacacgcg cctcgcatct tttaatgatg gaataatttg ggaatttact
     1921 ctgtatttat ttatttttat gttttgtatt tggattttag aaagtaaata aagaaggtag
     1981 aagagttacg gaatgaagaa aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc
     2041 gtactttaca tatatattta ttagacaaga aaagcagatt aaatagatat acattcgatt
     2101 aacgataagt aaaatgtaaa atcacaggat tttcgtgtgt ggtcttctac acagacaaga
     2161 tgaaacaatt cggcattaat acctgagagc aggaagagca agataaaagg tagtatttgt
     2221 tggcgatccc cctagagtct tttacatctt cggaaaacaa aaactatttt ttctttaatt
     2281 tcttttttta ctttctattt ttaatttata tatttatatt aaaaaattta aattataatt
     2341 atttttatag cacgtgatga aaaggaccct aagaaaccat tattatcatg acattaacct
     2401 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     2461 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     2521 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg cgattcggta atctccgaac
     2581 agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc atatgtagtg
     2641 ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca aaaacctgca
     2701 ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct actcatccta
     2761 gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt
     2821 cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta ggtcccaaaa
     2881 tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag ggcacagtta
     2941 agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaagac agaaaatttg
     3001 ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga atagcagaat
     3061 gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc ggtttgaagc
     3121 aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca gaattgtcat
     3181 gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt gcgaagagcg
     3241 acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga gatgaaggtt
     3301 acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagac gcattgggtc
     3361 aacagtatag aaccgtggat gatgtggtct ctacaggatc tgacattatt attgttggaa
     3421 gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac agaaaagcag
     3481 gctgggaagc atatttgaga agatgcggcc agcaaaacta agagctcgaa ttcatcgatg
     3541 atatcagatc cactagtggc ctatgcggcc gcggatctgc cggtctccct atagtgagtc
     3601 gtattaattt cgataagcca ggttaacctg cattaatgaa tcggccaacg cgcggggaga
     3661 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3721 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3781 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3841 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3901 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3961 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4021 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4081 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4141 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4201 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4261 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4321 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4381 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4441 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4501 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4561 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4621 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4681 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4741 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4801 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4861 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4921 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4981 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5041 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5101 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5161 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5221 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5281 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5341 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5401 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5461 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5521 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5581 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     5641 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcg
//
