LOCUS       sequence-405595-        2776 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 plasmid containing an eCFP 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-eCFP
FEATURES             Location/Qualifiers
     source          1..2776
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(565..567,568..570,571..1281)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(610..631)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(871..890)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1218..1239
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     rep_origin      complement(1426..2014)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1506..1525)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2105..2776,1..120))
                     /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 gccatccact gcggagccgt acaaatgtac ggccagcaac gtcggttcga gatggcgctc
       61 gatgacgcca actacctctg atagttgagt cgatacttcg gcgatcaccg cttccctcat
      121 gcgaaacgat cctcatcctg tctcttgatc agatattgat cccctgcgcc atcagatcct
      181 tggcggcaag aaagccatcc agtttacttt gcagggcttc ccaaccttac cagagggcgc
      241 cccagctggc aattccggtt cgcttgctaa gcttgcatgc ctgcaagtcg actctagagg
      301 agcggtgatc acaggcagca acgctctgtc atcgttacaa tcaacatgct accctccgcg
      361 agatcatccg tgtttcaaac ccggcagctt agttgccgtt cttccgaata gcatcggtaa
      421 catgagcaaa gtctgccgcc ttacaacggc tctcccgctg acgccgtccc ggactgatgg
      481 gctgcctgta tcgagtggtg attttgtgcc gagctcatta ctcgcatcca ttctcaggct
      541 gtctcgtctc gtctcaggtc tcaaatggtg agcaagggcg aggagctgtt caccggggtg
      601 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      661 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
      721 aagctgcccg tgccctggcc caccctcgtg accaccctga cctggggcgt gcagtgcttc
      781 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      841 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
      901 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
      961 gaggacggca acatcctggg gcacaagctg gagtacaact acatcagcca caacgtctat
     1021 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     1081 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1141 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     1201 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1261 ggcatggacg agctgtacaa gtcaggtcga gaccaccagg atacatagat taccacaact
     1321 ccgagccctt ccacccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1381 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1441 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1501 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1561 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1621 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1681 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1741 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1801 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1861 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1921 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1981 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2041 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatgc attctaggtg
     2101 attattattt gccgactacc ttggtgatct cgcctttcac gtagtggaca aattcttcca
     2161 actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc tgtctagctt
     2221 caagtatgac gggctgatac tgggccggca ggcgctccat tgcccagtcg gcagcgacat
     2281 ccttcggcgc gattttgccg gttactgcgc tgtaccaaat gcgggacaac gtaagcacta
     2341 catttcgctc atcgccagcc cagtcgggcg gcgagttcca tagcgttaag gtttcattta
     2401 gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc gctggaccta
     2461 ccaaggcaac gctatgttct cttgcttttg tcagcaagat agccagatca atgtcgatcg
     2521 tggctggctc gaagatacca gcaagaatgt cattgcgctg ccattctcca aattgcagtt
     2581 cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta
     2641 cagcgcggag aatctcactc tctccagggg aagccgaagt ttccaaaagg tcgttgatca
     2701 aagctcgccg cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac
     2761 tgtgtggctt caggcc
//
