LOCUS       sequence-468707-        5986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo-YTK; ConLS/RE cassette; Const. mNG;  pTDH3-mNeonGreen-tENO2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5986)
  AUTHORS   O'Laughlin R, Tran Q, Lezia A, Ngamkanjanarat W, Emmanuele P, Hao N,
            Hasty J
  TITLE     A Standardized Set of MoClo-Compatible Inducible Promoter Systems 
            for Tunable Gene Expression in Yeast.
  JOURNAL   ACS Synth Biol. 2024 Jan 19;13(1):85-102. doi: 
            10.1021/acssynbio.3c00184. Epub 2023 Dec 11.
  PUBMED    38079574
REFERENCE   2  (bases 1 to 5986)
  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"; doi: 
            "10.1021/acssynbio.3c00184"; journalName: "ACS Synth Biol"; date: 
            "2024-01-19- 19"; volume: "13"; issue: "1"; pages: "85-102"
COMMENT     Sequence Label: pRO211
FEATURES             Location/Qualifiers
     source          1..5986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      156..744
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     645..664
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1587..2253
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     2123..2146
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             3562..4365
                     /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"
     CDS             complement(join(5074..5865,5866..5934))
                     /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     5697..5716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5935..5986,1..53))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
       61 ccccgaaaag tgccacctgt catgaccaaa atcccttaac gtgagttttc gttccactga
      121 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
      181 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
      241 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
      301 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
      361 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
      421 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
      481 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
      541 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
      601 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
      661 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
      721 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
      781 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
      841 cgtgcggccg ccaatgctaa attcgagtga aacacaggaa gatcagaaaa tcctcatttc
      901 atccatatta acaataattt caaatgttta tttgcattat ttgaaactag gcaagacaag
      961 caacgaaacg tttttgaaaa ttttgagtat tttcaataaa tttgtagagg actcagatat
     1021 tgaaaaaaag ctacagcaat taatacttga taagaagagt attgagaagg gcaacggttc
     1081 atcatctcat ggatctgcac atgaacaaac accagagtca aacgacgttg aaattgaggc
     1141 tactgcgcca attgatgaca atacagacga tgataacaaa ccgaagttat ctgatgtaga
     1201 aaaggattaa agatgctaag agatagtgat gatatttcat aaataatgta attctatata
     1261 tgttaattac cttttttgcg aggcatattt atggtgaagg ataagttttg accatcaaag
     1321 aaggttaatg tggctgtggt ttcagggtcc ataaagccca catggataac attacccctg
     1381 aattcgcatc tagatggtag agccacaaac agccggtaca agcaacgatc tccaggacca
     1441 tctgaatcat gcgcggatga cacgaactca cgacggcgat cacagacatt aacccacagt
     1501 acagacactg cgacaacgtg gcaattcgtc gcaataccgt ctcactgaac tggccgataa
     1561 ttgcagacga acgcagttcg agtttatcat tatcaatact gccatttcaa agaatacgta
     1621 aataattaat agtagtgatt ttcctaactt tatttagtca aaaaattagc cttttaattc
     1681 tgctgtaacc cgtacatgcc caaaataggg ggcgggttac acagaatata taacatcgta
     1741 ggtgtctggg tgaacagttt attcctggca tccactaaat ataatggagc ccgcttttta
     1801 agctggcatc cagaaaaaaa aagaatccca gcaccaaaat attgttttct tcaccaacca
     1861 tcagttcata ggtccattct cttagcgcaa ctacagagaa caggggcaca aacaggcaaa
     1921 aaacgggcac aacctcaatg gagtgatgca acctgcctgg agtaaatgat gacacaaggc
     1981 aattgaccca cgcatgtatc tatctcattt tcttacacct tctattacct tctgctctct
     2041 ctgatttgga aaaagctgaa aaaaaaggtt gaaaccagtt ccctgaaatt attcccctac
     2101 ttgactaata agtatataaa gacggtaggt attgattgta attctgtaaa tctatttctt
     2161 aaacttctta aattctactt ttatagttag tctttttttt agttttaaaa caccaagaac
     2221 ttagtttcga ataaacacac ataaacaaac aaaagatcta tggtttcaaa aggggaggag
     2281 gataatatgg cgagtcttcc tgccacgcat gagctgcata tcttcggatc tattaatggt
     2341 gtcgactttg atatggtagg acaaggaacc gggaacccca atgacgggta tgaagagcta
     2401 aatctaaaga gtacaaaagg agatttacag ttttctcctt ggatacttgt tccccacatt
     2461 ggatatggat ttcatcaata tcttccttat ccagacggaa tgagcccgtt ccaggccgct
     2521 atggttgacg gctccgggta tcaagtacac cgtaccatgc agtttgagga tggggcatcc
     2581 ctgaccgtaa attacaggta cacctacgag ggctcacaca taaagggtga ggcgcaggta
     2641 aagggtactg gatttcctgc ggacggccca gtaatgacaa attcacttac agcagcggat
     2701 tggtgcagga gcaaaaagac ctatcccaat gacaaaacta tcataagtac ctttaaatgg
     2761 tcctatacga caggtaacgg gaagagatat aggtcaacgg caaggacaac gtatactttc
     2821 gcgaagccaa tggcagcaaa ctatctgaag aatcaaccta tgtacgtgtt cagaaagacc
     2881 gagttgaaac attccaagac agagttaaat ttcaaagaat ggcaaaaggc ttttactgac
     2941 gtaatgggga tggatgaatt gtataaggga tcctaactcg agagtgcttt taactaagaa
     3001 ttattagtct tttctgctta ttttttcatc atagtttaga acactttata ttaacgaata
     3061 gtttatgaat ctatttaggt ttaaaaattg atacagtttt ataagttact ttttcaaaga
     3121 ctcgtgctgt ctattgcata atgcactgga aggggaaaaa aaaggtgcac acgcgtggct
     3181 ttttcttgaa tttgcagttt gaaaaatgct gccaatgaga cgacggggtc atcacggctc
     3241 atcatgcgcc aaacaaatgt gtgcaataca cgctcggatg actgcatgat gaccgcactg
     3301 actggggaca gcagatccac ctaagcctgt gagagaagca gacacccgac agatcaaggc
     3361 agttaactag tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc
     3421 gaacagaagg aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt
     3481 agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg
     3541 tgcaggaaac gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat
     3601 cctagtcctg ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt
     3661 gcttcattgg atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc
     3721 aaaatttgtt tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca
     3781 gttaagccgc taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa
     3841 tttgctgaca ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca
     3901 gaatgggcag acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg
     3961 aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg
     4021 tcatgcaagg gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag
     4081 agcgacaaag attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa
     4141 ggttacgatt ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg
     4201 ggtcaacagt atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt
     4261 ggaagaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa
     4321 gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa
     4381 gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta
     4441 cccgagtaga gcacttgaat ccactgcccc gggaatctcg gtcgtaatga tttctataat
     4501 gacgaaaaaa aaaaaattgg aaagaaaaag cttcatggcc tttataaaaa ggaactatcc
     4561 aatacctcgc cagaaccaag taacagtatt ttacggggca caaatcaaga acaataagac
     4621 aggactgtaa agatggacgc attgaactcc aaagaacaac aagagttcca aaaagtagtg
     4681 gaacaaaagc aaatgaagga tttcatgcgt ttgtactcta atctggtaga aagatgtttc
     4741 acagactgtg tcaatgactt cacaacatca aagctaacca ataaggaaca aacatgcatc
     4801 atgaagtgct cagaaaagtt cttgaagcat agcgaacgtg tagggcagcg tttccaagaa
     4861 caaaacgctg ccttgggaca aggcttgggc cgataaggtg tactggcgta tatatatcta
     4921 attatgtatc tctggtgtag cccattttta gcatgtaaat ataaagaccg agcggccgcg
     4981 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5041 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5101 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5161 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgggaccc
     5221 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5281 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5341 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5401 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5461 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5521 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5581 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5641 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5701 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5761 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5821 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5881 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5941 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatg
//
