LOCUS       sequence-374549-        1729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2A peptide P2A cloned in yeast MoClo entry vector (pYTK001) for use 
            as the first 2A peptide in quad-cistronic construct assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1729)
  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 1729)
  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_P2Aq(1st)
FEATURES             Location/Qualifiers
     source          1..1729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     336..355
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             555..611
                     /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(630..664)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(735..1394)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1309..1328
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      join(1576..1729,1..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 ggtctcggag cggtgcaaca aatttctcat tgttgaagca ggcgggagat gtggaggaga
      601 accctggacc taggagaccg accagaccaa taaaaaacgc ccggcggcaa ccgagcgttc
      661 tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg
      721 agctcgatat caaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta
      781 agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc
      841 atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag
      901 ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgaa
      961 acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac
     1021 gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag
     1081 agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc
     1141 catatcacca gctcaccgtc tttcattgcc atacgaaatt ccggatgagc attcatcagg
     1201 cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt
     1261 aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga
     1321 aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg
     1381 atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg
     1441 cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg cccgatcaat
     1501 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1561 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1621 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1681 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttcta
//
