LOCUS       sequence-421776-        2376 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes mRuby2 fluorescent protein as a Type 3b' part to be used in 
            the yeast secretion/display (YSD) extension of the MoClo yeast 
            toolkit (YTK).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2376)
  AUTHORS   O'Riordan NM, Juric V, O'Neill SK, Roche AP, Young PW
  TITLE     A Yeast Modular Cloning (MoClo) Toolkit Expansion for Optimization 
            of Heterologous Protein Secretion and Surface Display in 
            Saccharomyces cerevisiae.
  JOURNAL   ACS Synth Biol. 2024 Mar 14. doi: 10.1021/acssynbio.3c00743.
  PUBMED    38483353
REFERENCE   2  (bases 1 to 2376)
  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: "ACS Synth 
            Biol. 2024 Mar 14. doi: 10.1021/acssynbio.3c00743."
COMMENT     Sequence Label: pYSD063
FEATURES             Location/Qualifiers
     source          1..2376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     222..241
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      489..1077
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     978..997
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1192..1899
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTHM
                     ALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAVA
                     KFAGLGGGMDELYK"
     terminator      complement(1919..1953)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(join(2024..2376,1..307))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
       61 ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa
      121 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
      181 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
      241 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
      301 tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
      361 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgcccgatc aatcatgacc
      421 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
      481 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
      541 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
      601 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
      661 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
      721 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
      781 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
      841 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
      901 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
      961 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     1021 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     1081 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     1141 tttcctgcgt tatcccctga ttctgtggat aaccgtagtc ggtctcatgc tgtgtccaaa
     1201 ggagaggagt taatcaagga aaacatgaga atgaaagttg tcatggaggg ctccgttaat
     1261 ggtcaccaat tcaagtgtac aggggaaggt gaaggtaatc cttacatggg tacacaaact
     1321 atgagaatta aagtaattga aggcggacca ctaccatttg catttgacat tctggcaacg
     1381 tcattcatgt acggatcacg aactttcatc aagtacccta aaggtatacc agactttttc
     1441 aagcaatctt ttccagaggg ttttacatgg gaaagggtta caagatacga agatgggggt
     1501 gtcgtcacag ttatgcaaga tacttcatta gaagatggct gccttgtcta tcatgtgcaa
     1561 gtaagagggg tgaattttcc ttctaacgga cctgtgatgc agaaaaagac caaaggttgg
     1621 gaaccaaata ctgaaatgat gtacccagct gatggaggtt tgagaggcta cacacacatg
     1681 gcgcttaaag ttgatggtgg aggtcatttg tcttgtagtt ttgttaccac ttatcgttct
     1741 aaaaagactg ttggcaatat caaaatgcca ggaatacatg ctgtagacca cagactagaa
     1801 agactcgaag agagcgataa cgaaatgttc gttgtacaga gagagcatgc cgtagccaaa
     1861 tttgctggct taggcggtgg tatggatgaa ttgtataagg gatcctgaga ccagaccaat
     1921 aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat
     1981 tactggatct atcaacagga gtccaagcga gctcgatatc aaattacgcc ccgccctgcc
     2041 actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa
     2101 cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt
     2161 tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac
     2221 tggtgaaact cacccaggga ttggctgaaa cgaaaaacat attctcaata aaccctttag
     2281 ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact
     2341 gccggaaatc gtcgtggtat tcactccaga gcgatg
//
