LOCUS       sequence-405611-        2776 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 plasmid containing an eGFP 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-eGFP
FEATURES             Location/Qualifiers
     source          1..2776
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(509..1300)
                     /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"
     CDS             join(1745..1747,1748..1750,1751..2461)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1745..1747,1748..1750,1751..2461)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1790..1811)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2051..2070)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2398..2419
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     rep_origin      complement(join(2606..2776,1..418))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2686..2705)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
       61 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
      121 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
      181 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
      241 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
      301 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
      361 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
      421 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
      481 gattttggtc atgcattcta ggtgattatt atttgccgac taccttggtg atctcgcctt
      541 tcacgtagtg gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt
      601 gtccaagata agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct
      661 ccattgccca gtcggcagcg acatccttcg gcgcgatttt gccggttact gcgctgtacc
      721 aaatgcggga caacgtaagc actacatttc gctcatcgcc agcccagtcg ggcggcgagt
      781 tccatagcgt taaggtttca tttagcgcct caaatagatc ctgttcagga accggatcaa
      841 agagttcctc cgccgctgga cctaccaagg caacgctatg ttctcttgct tttgtcagca
      901 agatagccag atcaatgtcg atcgtggctg gctcgaagat accagcaaga atgtcattgc
      961 gctgccattc tccaaattgc agttcgcgct tagctggata acgccacgga atgatgtcgt
     1021 cgtgcacaac aatggtgact tctacagcgc ggagaatctc actctctcca ggggaagccg
     1081 aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc cttacggtca
     1141 ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc gccatccact gcggagccgt
     1201 acaaatgtac ggccagcaac gtcggttcga gatggcgctc gatgacgcca actacctctg
     1261 atagttgagt cgatacttcg gcgatcaccg cttccctcat gcgaaacgat cctcatcctg
     1321 tctcttgatc agatattgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc
     1381 agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccggtt
     1441 cgcttgctaa gcttgcatgc ctgcaagtcg actctagagg agcggtgatc acaggcagca
     1501 acgctctgtc atcgttacaa tcaacatgct accctccgcg agatcatccg tgtttcaaac
     1561 ccggcagctt agttgccgtt cttccgaata gcatcggtaa catgagcaaa gtctgccgcc
     1621 ttacaacggc tctcccgctg acgccgtccc ggactgatgg gctgcctgta tcgagtggtg
     1681 attttgtgcc gagctcatta ctcgcatcca ttctcaggct gtctcgtctc gtctcaggtc
     1741 tcaaatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1801 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1861 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1921 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     1981 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2041 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2101 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2161 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     2221 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     2281 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     2341 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     2401 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     2461 gtcaggtcga gaccaccagg acacatagat taccacaact ccgagccctt ccacccacag
     2521 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2581 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2641 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2701 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2761 tgtccgcctt tctccc
//
