LOCUS       sequence-405588-        2776 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 plasmid containing an eYFP fluorescent protein with CD 
            overhangs used to build a level 1 construct..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2776)
  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 2776)
  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-eYFP
FEATURES             Location/Qualifiers
     source          1..2776
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1067..1069,1070..1072,1073..1783)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1112..1133)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1373..1392)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1720..1741
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     rep_origin      complement(1928..2516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2008..2027)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2607..2776,1..622))
                     /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 cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa
       61 gcactacatt tcgctcatcg ccagcccagt cgggcggcga gttccatagc gttaaggttt
      121 catttagcgc ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg
      181 gacctaccaa ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt
      241 cgatcgtggc tggctcgaag ataccagcaa gaatgtcatt gcgctgccat tctccaaatt
      301 gcagttcgcg cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga
      361 cttctacagc gcggagaatc tcactctctc caggggaagc cgaagtttcc aaaaggtcgt
      421 tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa
      481 tatcactgtg tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt acggccagca
      541 acgtcggttc gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt
      601 cggcgatcac cgcttccctc atgcgaaacg atcctcatcc tgtctcttga tcagatattg
      661 atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact ttgcagggct
      721 tcccaacctt accagagggc gccccagctg gcaattccgg ttcgcttgct aagcttgcat
      781 gcctgcaagt cgactctaga ggagcggtga tcacaggcag caacgctctg tcatcgttac
      841 aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc ttagttgccg
      901 ttcttccgaa tagcatcggt aacatgagca aagtctgccg ccttacaacg gctctcccgc
      961 tgacgccgtc ccggactgat gggctgcctg tatcgagtgg tgattttgtg ccgagctcat
     1021 tactcgcatc cattctcagg ctgtctcgtc tcgtctcagg tctcaaatgg tgagcaaggg
     1081 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     1141 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     1201 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt
     1261 cggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     1321 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1381 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1441 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1501 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     1561 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     1621 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
     1681 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1741 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtcaggtc gagaccacca
     1801 ggatacatag attaccacaa ctccgagccc ttccacccac agaatcaggg gataacgcag
     1861 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1921 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1981 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2041 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2101 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2161 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2221 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2281 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2341 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2401 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2461 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     2521 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     2581 ttttggtcat gcattctagg tgattattat ttgccgacta ccttggtgat ctcgcctttc
     2641 acgtagtgga caaattcttc caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt
     2701 ccaagataag cctgtctagc ttcaagtatg acgggctgat actgggccgg caggcgctcc
     2761 attgcccagt cggcag
//
