LOCUS       sequence-189621-        1874 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  part designed to occupy position 20 of EMMA. Functional category: 
            p2A sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1874)
  AUTHORS   Martella A, Matjusaitis M, Auxillos J, Pollard SM, Cai Y
  TITLE     EMMA: An Extensible Mammalian Modular Assembly Toolkit for the Rapid
            Design and Production of Diverse Expression Vectors.
  JOURNAL   ACS Synth Biol. 2017 Apr 24. doi: 10.1021/acssynbio.7b00016.
  PUBMED    28418644
REFERENCE   2  (bases 1 to 1874)
  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. 2017 Apr 24. doi: 10.1021/acssynbio.7b00016."
COMMENT     Sequence Label: YCe2064 HC_Kan_p2A_p20
FEATURES             Location/Qualifiers
     source          1..1874
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             81..137
                     /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      217..248
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        249..351
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             352..1167
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(425..444)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1191..1218
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1230..1817)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1310..1329)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1839..1868)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 ttgattgcag tccagttacg ctggagtctg aggctcgtcc tgaatgatat caagcttgaa
       61 ttcgttacgt ctcgagcggc gctactaact tcagcctgct gaagcaggct ggcgacgtgg
      121 aggagaaccc tggaccttct ggacgagacg aagacgaatt ctctagatat cgctcaatac
      181 tgaccattta aatcatacct gacctccata gcagaaagtc aaaagcctcc gaccggaggc
      241 ttttgacttg atcggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta
      301 ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa aatgagccat
      361 attcaacggg aaacgtcttg ctcgaggccg cgattaaatt ccaacatgga tgctgattta
      421 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg
      481 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
      541 gatgttacag atgagatggt caggctaaac tggctgacgg aatttatgcc tcttccgacc
      601 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatcccaggg
      661 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
      721 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacggc
      781 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttggtgcg
      841 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
      901 aagcttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
      961 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     1021 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     1081 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     1141 cacttgatgc tcgatgagtt tttctaatga gggcccaaat gtaatcacct ggctcacctt
     1201 cgggtgggcc tttctgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1261 cacaaaaatc gatgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1321 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1381 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1441 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1501 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1561 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1621 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     1681 ggtatctgcg ctctgctgaa gccagttacc tcggaaaaag agttggtagc tcttgatccg
     1741 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1801 gaaaaaaagg atctcaagaa gatcctttga ttttctaccg aagaaaggcc cacccgtgaa
     1861 ggtgagccag tgag
//
