LOCUS       sequence-192919-        1871 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 1871)
  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 1871)
  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..1871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             82..138
                     /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      218..249
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        250..352
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             353..1168
                     /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(426..445)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1189..1216
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1228..1815)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1308..1327)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1837..1866)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 gttgattgca gtccagttac gctggagtct gaggctcgtc ctgaatgata tcaagcttga
       61 attcgttacg tctcgagcgg cgctactaac ttcagcctgc tgaagcaggc tggcgacgtg
      121 gaggagaacc ctggaccttc tggacgagac gaagacgaat tctctagata tcgctcaata
      181 ctgaccattt aaatcatacc tgacctccat agcagaaagt caaaagcctc cgaccggagg
      241 cttttgactt gatcggcacg taagaggttc caactttcac cataatgaaa taagatcact
      301 accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatgagcca
      361 tattcaacgg gaaacgtctt gctcgaggcc gcgattaaat tccaacatgg atgctgattt
      421 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt
      481 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
      541 tgatgttaca gatgagatgg tcaggctaaa ctggctgacg gaatttatgc ctcttccgac
      601 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatcccagg
      661 gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
      721 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacgg
      781 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttggtgc
      841 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
      901 taagcttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
      961 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     1021 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     1081 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     1141 tcacttgatg ctcgatgagt ttttctgagg gcccaaatgt aatcacctgg ctcaccttcg
     1201 ggtgggcctt tctgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     1261 caaaaatcga tgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     1321 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     1381 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     1441 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     1501 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     1561 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     1621 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     1681 tatctgcgct ctgctgaagc cagttacctc ggaaaaagag ttggtagctc ttgatccggc
     1741 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     1801 aaaaaaggat ctcaagaaga tcctttgatt ttctaccgaa gaaaggccca cccgtgaagg
     1861 tgagccagtg a
//
