LOCUS       sequence-405586-        2193 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 plasmid containing a dual 2A (P2A and T2A) peptide sequence 
            with CD overhangs used to build level 1 constructs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2193)
  AUTHORS   Pollak B, Matute T, Nunez I, Cerda A, Lopez C, Vargas V, Kan A, 
            Bielinski V, von Dassow P, Dupont CL, Federici F
  TITLE     Universal loop assembly: open, efficient and cross-kingdom DNA 
            fabrication.
  JOURNAL   Synth Biol (Oxf). 2020;5(1):ysaa001. doi: 10.1093/synbio/ysaa001. 
            Epub 2020 Feb 5.
  PUBMED    32161816
REFERENCE   2  (bases 1 to 2193)
  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: "Synth Biol 
            (Oxf)."; date: "2020"; volume: "5(1):ysaa001. doi"; pages: " 
            10.1093/synbio/ysaa001. Epub 2020 Feb 5"
COMMENT     Sequence Label: pL0_CD-PT2A
FEATURES             Location/Qualifiers
     source          1..2193
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      777..1365
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1266..1285
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1510..1563)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(1573..1629)
                     /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"
     CDS             join(2088..2193,1..686)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg
       61 taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt
      121 cccctggaga gagtgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca
      181 tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg
      241 acattcttgc tggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga
      301 caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc
      361 cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc
      421 cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca
      481 gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc
      541 tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag
      601 aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg
      661 agatcaccaa ggtagtcggc aaataataat cacctagaat gcatgaccaa aatcccttaa
      721 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
      781 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
      841 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
      901 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
      961 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1021 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1081 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1141 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1201 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1261 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1321 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1381 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1441 tcccctgatt ctgtgggtgg aagggctcgg agttgtggta atctatgtat cctggtggtc
     1501 tcgacctgat gggccgggat tttcctccac gtccccgcat gttagaagac ttcccctgcc
     1561 ctcgccggag ccaggcccgg ggttttcttc aacatcacct gcttgcttta gcagagagaa
     1621 gtttgtggcg ccgctgcccc catttgagac ctgagacgag acgagacagc ctgagaatgg
     1681 atgcgagtaa tgagctcggc acaaaatcac cactcgatac aggcagccca tcagtccggg
     1741 acggcgtcag cgggagagcc gttgtaaggc ggcagacttt gctcatgtta ccgatgctat
     1801 tcggaagaac ggcaactaag ctgccgggtt tgaaacacgg atgatctcgc ggagggtagc
     1861 atgttgattg taacgatgac agagcgttgc tgcctgtgat caccgctcct ctagagtcga
     1921 cttgcaggca tgcaagctta gcaagcgaac cggaattgcc agctggggcg ccctctggta
     1981 aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc
     2041 gcaggggatc aatatctgat caagagacag gatgaggatc gtttcgcatg agggaagcgg
     2101 tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag cgccatctcg
     2161 aaccgacgtt gctggccgta catttgtacg gct
//
