LOCUS       sequence-421766-        1926 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the wildtype S. cerevisiae MFalpha1 pre-pro- signal peptide 
            (MFappWT) as a Type 3a' 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 1926)
  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 1926)
  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: pYSD024
FEATURES             Location/Qualifiers
     source          1..1926
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(11..67,68..265,266..277)
                     /codon_start=1
                     /gene="MF-alpha-1"
                     /product="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /label=alpha-factor secretion signal
                     /note="Cleavage by the Kex2 protease occurs after the 
                     dibasic KR sequence. The EA dipeptides are then removed by 
                     dipeptidyl aminopeptidase A."
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYLDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLDKREADA"
     terminator      complement(291..325)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(396..1055)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     970..989
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1237..1825
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1726..1745
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcggtctcat atgagatttc cttcaatttt tactgcagtt ttattcgcag catcctccgc
       61 attagctgct ccagtcaaca ctacaacaga agatgaaacg gcacaaattc cggctgaagc
      121 tgtcatcggt tacttagatt tagaagggga tttcgatgtt gctgttttgc cattttccaa
      181 cagcacaaat aacgggttat tgtttataaa tactactatt gccagcattg ctgctaaaga
      241 agaaggggta tctttggata aaagagaggc tgatgcttga gaccagacca ataaaaaacg
      301 cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat
      361 ctatcaacag gagtccaagc gagctcgata tcaaattacg ccccgccctg ccactcatcg
      421 cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga
      481 tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg
      541 gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa
      601 ctcacccagg gattggctga aacgaaaaac atattctcaa taaacccttt agggaaatag
      661 gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa
      721 tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg
      781 taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc catacgaaat
      841 tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc
      901 ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag
      961 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata
     1021 tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat
     1081 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa
     1141 cctcttacgt gcccgatcaa tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     1201 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1261 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     1321 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     1381 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     1441 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     1501 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     1561 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     1621 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     1681 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     1741 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     1801 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     1861 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     1921 ccgtag
//
