LOCUS       sequence-367243-        1730 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2A peptide P2A cloned in yeast MoClo entry vector (pYTK001) for use 
            in bi-cistronic and as the first 2A peptide in tri-cistronic 
            construct assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1730)
  AUTHORS   Mukherjee M, Wang ZQ
  TITLE     A Well-Characterized Polycistronic-Like Gene Expression System in 
            Yeast.
  JOURNAL   Biotechnol Bioeng. 2022 Sep 27. doi: 10.1002/bit.28247.
  PUBMED    36168285
REFERENCE   2  (bases 1 to 1730)
  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. 2022 Sep 27. doi: 10.1002/bit.28247."
COMMENT     Sequence Label: pYTK001_P2Ab
FEATURES             Location/Qualifiers
     source          1..1730
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     336..355
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             556..612
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     terminator      complement(631..665)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(736..1395)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1310..1329
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      join(1577..1730,1..435)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
       61 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
      121 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
      181 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
      241 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
      301 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
      361 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
      421 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
      481 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtagtcgg
      541 ggtctcggtt ctggtgcaac aaatttctca ttgttgaagc aggcgggaga tgtggaggag
      601 aaccctggac ctaggagacc gaccagacca ataaaaaacg cccggcggca accgagcgtt
      661 ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc
      721 gagctcgata tcaaattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt
      781 aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg
      841 catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa
      901 gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga
      961 aacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca
     1021 cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca
     1081 gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc
     1141 ccatatcacc agctcaccgt ctttcattgc catacgaaat tccggatgag cattcatcag
     1201 gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt
     1261 taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg
     1321 aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt
     1381 gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
     1441 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gcccgatcaa
     1501 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1561 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1621 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1681 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
//
