LOCUS       sequence-259429-        6754 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LEU2 integration vector of pZF(3BS)-mRUBY2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6754)
  AUTHORS   An-Adirekkun JM, Stewart CJ, Geller SH, Patel MT, Melendez J, Oakes 
            BL, Noyes MB, McClean MN
  TITLE     A yeast optogenetic toolkit (yOTK) for gene expression control in 
            Saccharomyces cerevisiae.
  JOURNAL   Biotechnol Bioeng. 2019 Dec 2. doi: 10.1002/bit.27234.
  PUBMED    31788779
REFERENCE   2  (bases 1 to 6754)
  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"; journalName: "Biotechnol 
            Bioeng. 2019 Dec 2. doi: 10.1002/bit.27234."
COMMENT     Sequence Label: pMM0553
FEATURES             Location/Qualifiers
     source          1..6754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1404..1427
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1451..2161
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     terminator      2177..2364
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2232..2256)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2819..3226
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             3227..4321
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     CDS             complement(5049..5864)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5049..5864)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5772..5791
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5856..5875
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6089..6677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6578..6597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgattct gtggataacc gtgcggccgc caatacccag aagaacagta aaataaagca
       61 aggtacgtga aattaatatt tttaaatggt tctaaccgat gccgaagaac tgcgcagtcc
      121 ggttataacg tctgacatgt ccttttttga tttggaatcc aaccactcaa gtgactctgt
      181 tcatttactt tgcgaaaaat atacccacaa attgcccatc gaaagtgaat cgcaaaccac
      241 cttcagactg gcaccgacaa agcaaagatt atacagacag agtactttat acgtaccgtt
      301 aagtctcaag caaagggttt tcttatttac tgaacgggta aagagtatct gggccggctt
      361 gccaagatgc aaaccgaata agtatttcaa agttgcattt gccttagccg tcctgacacc
      421 attggctatt tggatatttt atattgactt tcgtgtacat tgatcacatc gactgttcta
      481 ttggcaaatg aaccacgggc attgactatt tttcaggtta ctactatata ttatcatcac
      541 gggcaaggat tgtaccctga attcgcatct agatggtaga gccacaaaca gccggtacaa
      601 gcaacgatct ccaggaccat ctgaatcatg cgcggatgac acgaactcac gacggcgatc
      661 acagacatta acccacagta cagacactgc gacaacgtgg caattcgtcg caataccgtc
      721 tcactgaact ggccgataat tgcagacgaa cgcgaggcaa gctaaacaga tctttatatt
      781 gaattttcaa aaattcttac tttttttttg gatggacgca aagaagttta ataatcatat
      841 tacatggcat taccaccata tacatatcca tatacatatc catatctaat cttacttata
      901 tgttgtggaa atgtaaagag ccccattatc ttagcctaaa aaaaccttct ctttggaact
      961 ttcagtaata cgcttaactg ctcattgcta tattgaagtg cgcgcgcgcg tgggcgagcg
     1021 tgggcgagcg tgggcgtcta gaccgtgcgt cctcgtcttc accggtcgcg ttcctgaaac
     1081 gcagatgtgc ctcgcgccgc actgctccga acaataaaga ttctacaata ctagctttta
     1141 tggttatgaa gaggaaaaat tggcagtaac ctggccccac aaaccttcaa attaacgaat
     1201 caaattaaca accataggat gataatgcga ttagtttttt agccttattt ctggggtaat
     1261 taatcagcga agcgatgatt tttgatctat taacagatat ataaatggaa aagctgcata
     1321 accactttaa ctaatacttt caacattttc agtttgtatt acttcttatt caaatgtcat
     1381 aaaagtatca acaaaaaatt gttaatatac ctctatactt taacgtcaag gagaaaaaac
     1441 tataggtact atggtgtcca aaggagagga gttaatcaag gaaaacatga gaatgaaagt
     1501 tgtcatggag ggctccgtta atggtcacca attcaagtgt acaggggaag gtgaaggtaa
     1561 tccttacatg ggtacacaaa ctatgagaat taaagtaatt gaaggcggac cactaccatt
     1621 tgcatttgac attctggcaa cgtcattcat gtacggatca cgaactttca tcaagtaccc
     1681 taaaggtata ccagactttt tcaagcaatc ttttccagag ggttttacat gggaaagggt
     1741 tacaagatac gaagatgggg gtgtcgtcac agttatgcaa gatacttcat tagaagatgg
     1801 ctgccttgtc tatcatgtgc aagtaagagg ggtgaatttt ccttctaacg gacctgtgat
     1861 gcagaaaaag accaaaggtt gggaaccaaa tactgaaatg atgtacccag ctgatggagg
     1921 tttgagaggc tacacacaca tggcgcttaa agttgatggt ggaggtcatt tgtcttgtag
     1981 ttttgttacc acttatcgtt ctaaaaagac tgttggcaat atcaaaatgc caggaataca
     2041 tgctgtagac cacagactag aaagactcga agagagcgat aacgaaatgt tcgttgtaca
     2101 gagagagcat gccgtagcca aatttgctgg cttaggcggt ggtatggatg aattgtataa
     2161 gggatcctaa ctcgaggcga atttcttatg atttatgatt tttattatta aataagttat
     2221 aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt
     2281 attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagc atgaggtcgc
     2341 tcttattgac cacacctcta ccggcatgcc gagcaaatgc ctgcaaatcg ctccccattt
     2401 cgctgccaat gagacgacgg ggtcatcacg gctcatcatg cgccaaacaa atgtgtgcaa
     2461 tacacgctcg gatgactgca tgatgaccgc actgactggg gacagcagat ccacctaagc
     2521 ctgtgagaga agcagacacc cgacagatca aggcagttaa ctagtgcact gcagtacatc
     2581 gaggagaact tctagtatat ctacatacct aatattattg ccttattaaa aatggaatcc
     2641 caacaattac atcaaaatcc acattctctt caaaatcaat tgtcctgtac ttccttgttc
     2701 atgtgtgttc aaaaacgtta tatttatagg ataattatac tctatttctc aacaagtaat
     2761 tggttgtttg gccgagcggt ctaaggcgcc tgattcaaga aatatcttga ccgcagttaa
     2821 ctgtgggaat actcaggtat cgtaagatgc aagagttcga atctcttagc aaccattatt
     2881 tttttcctca acataacgag aacacacagg ggcgctatcg cacagaatca aattcgatga
     2941 ctggaaattt tttgttaatt tcagaggtcg cctgacgcat ataccttttt caactgaaaa
     3001 attgggagaa aaaggaaagg tgagagcgcc ggaaccggct tttcatatag aatagagaag
     3061 cgttcatgac taaatgcttg catcacaata cttgaagttg acaatattat ttaaggacct
     3121 attgtttttt ccaataggtg gttagcaatc gtcttacttt ctaacttttc ttacctttta
     3181 catttcagca atatatatat atatatttca aggatatacc attctaatgt ctgcccctaa
     3241 gaagatcgtc gttttgccag gtgaccacgt tggtcaagaa atcacagccg aagccattaa
     3301 ggttcttaaa gctatttctg atgttcgttc caatgtcaag ttcgatttcg aaaatcattt
     3361 aattggtggt gctgctatcg atgctacagg tgttccactt ccagatgagg cgctggaagc
     3421 ctccaagaag gctgatgccg ttttgttagg tgctgtgggt ggtcctaaat ggggaaccgg
     3481 tagtgttaga cctgaacaag gtttactaaa aatccgtaaa gaacttcaat tgtacgccaa
     3541 cttaagacca tgtaactttg catccgactc tcttttagac ttatctccaa tcaagccaca
     3601 atttgctaaa ggtactgact tcgttgttgt cagagaatta gtgggaggta tttactttgg
     3661 taagagaaag gaggacgatg gtgatggtgt cgcttgggat agtgaacaat acaccgttcc
     3721 agaagtgcaa agaatcacaa gaatggccgc tttcatggcc ctacaacatg agccaccatt
     3781 gcctatttgg tccttggata aagctaatgt tttggcctct tcaagattat ggagaaaaac
     3841 tgtggaggaa accatcaaga acgaatttcc tacattgaag gttcaacatc aattgattga
     3901 ttctgccgcc atgatcctag ttaagaaccc aacccaccta aatggtatta taatcaccag
     3961 caacatgttt ggtgatatca tctccgatga agcctccgtt atcccaggtt ccttgggttt
     4021 gttgccatct gcgtccttgg cctctttgcc agacaagaac accgcatttg gtttgtacga
     4081 accatgccac ggttctgctc cagatttgcc aaagaataag gtcaacccta tcgccactat
     4141 cttgtctgct gcaatgatgt tgaaattgtc attgaacttg cctgaagaag gtaaggccat
     4201 tgaagatgca gttaaaaagg ttttggatgc aggtatcaga actggtgatt taggtggttc
     4261 caacagtacc accgaagtcg gtgatgctgt cgccgaagaa gttaagaaaa tccttgctta
     4321 acctggagga cccttctctt tagactattc tactcttatg cacgtaaaaa attctaggaa
     4381 atatgtatta actaggagta aaataaccgg ctagtggcat tcatatagcc gtctgtttac
     4441 atctacatca cacatttcga gtgtatatct cgcaacgttg gcgttaaata ggcaggagta
     4501 ctcgtatcgc atgtcggtgc gacacgaaat tacaaaatgg aatatgttca tagggtagac
     4561 gaaactatat acgcaatcta catacattta tcaagaagga gaaaaaggag gatgtaaagg
     4621 aatacaggta agcaaattga tactaatggc tcaacgtgat aaggaaaaag aattgcactt
     4681 taacattaat attgacaagg aggagggcac cacacaaaaa gttaggtgta acagaaaatc
     4741 atgaaactat gattcctaat ttatatattg gaggattttc tctaaaaaaa aaaaaataca
     4801 acaaataaaa aacactcaat gacctgacca tttgatggag tttaagtcaa taccttcttg
     4861 aaccatttcc cataatggtg aaagttccct caagaatttt actctgtcag aaacggcctt
     4921 aacgacgtag tcgacctcct cttcagtact aaatctacca ataccaaatc tgatggaaga
     4981 atgggctaat gcatcatcct tacccagcgc cgagcggccg cgtgttacaa ccaattaacc
     5041 aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     5101 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     5161 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     5221 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     5281 gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca gacttgttca
     5341 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     5401 cgtgattgcg cctgagcgag gcgaaatacg cgatcgctgt taaaaggaca attacaaaca
     5461 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     5521 tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac
     5581 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     5641 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     5701 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     5761 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     5821 aatcgcggcc tggagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta
     5881 ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg
     5941 taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag
     6001 ttgaaggatc agtcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6061 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6121 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6181 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6241 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6301 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6361 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6421 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6481 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6541 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6601 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6661 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6721 ggccttttgc tcacatgttc tttcctgcgt tatc
//
