LOCUS       sequence-366231-        1723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2A peptide O2A cloned in yeast MoClo entry vector (pYTK001) for use 
            as the second 2A peptide in tri-cistronic and third 2A peptide in 
            quad-cistronic construct assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1723)
  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 1723)
  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_O2At(2nd)
FEATURES             Location/Qualifiers
     source          1..1723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     336..355
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(624..658)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(729..1388)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1303..1322
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      join(1570..1723,1..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 ggtctcttcc ggtaagtcca attacgacct attgaagttg tgtggagatg ttgaatccaa
      601 cccaggggag accgaccaga ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca
      661 aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg
      721 atatcaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
      781 ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc
      841 accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc
      901 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgaaacgaaa
      961 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
     1021 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
     1081 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
     1141 accagctcac cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca
     1201 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
     1261 gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc
     1321 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
     1381 ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt
     1441 agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgcccgat caatcatgac
     1501 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1561 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1621 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1681 aactggcttc agcagagcgc agataccaaa tactgttctt cta
//
