LOCUS       sequence-459780-        2979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lvl 0 5 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2979)
  AUTHORS   Duportet X, Wroblewska L, Guye P, Li Y, Eyquem J, Rieders J, 
            Rimchala T, Batt G, Weiss R
  TITLE     A platform for rapid prototyping of synthetic gene networks in 
            mammalian cells.
  JOURNAL   Nucleic Acids Res. 2014 Dec 1;42(21):13440-51. doi: 
            10.1093/nar/gku1082. Epub 2014 Nov 5.
  PUBMED    25378321
REFERENCE   2  (bases 1 to 2979)
  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"; doi: 
            "10.1093/nar/gku1082"; journalName: "Nucleic Acids Res"; date: 
            "2014-12-1- 1"; volume: "42"; issue: "21"; pages: "13440-51"
COMMENT     Sequence Label: lvl0-G-eBFP2
FEATURES             Location/Qualifiers
     source          1..2979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     regulatory      841..850
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(847..849,850..852,853..1566)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(892..913)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1606..1624)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1692..1796
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2018..2809
                     /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 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tcactctgtg gtctcaaggt
      841 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      901 ctggacggcg acgtaaacgg ccacaagttc agcgtgaggg gcgagggcga gggcgatgcc
      961 accaacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     1021 cccaccctcg tgaccaccct gagccacggc gtgcagtgct tcgcccgcta ccccgaccac
     1081 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     1141 atcttcttca aggacgacgg cacctacaag acccgcgccg aggtgaagtt cgagggcgac
     1201 accctagtga accgcatcga gctgaagggc gtcgacttca aggaggacgg caacatcctg
     1261 gggcacaagc tggagtacaa cttcaacagc cacaacatct atatcatggc cgtcaagcag
     1321 aagaacggca tcaaggtgaa cttcaagatc cgccacaacg tggaggacgg cagcgtgcag
     1381 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1441 agccactacc tgagcaccca gtccgtgctg agcaaagacc ccaacgagaa gcgcgatcac
     1501 atggtcctgc tggagttccg caccgccgcc gggatcactc tcggcatgga cgagctgtac
     1561 aagtaagctt tgagaccacg aagtggctct tcagtggacg aaagggcctc gtgatacgcc
     1621 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     1681 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1741 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatgc
     1801 gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg
     1861 ttttcatggc ttgttatgac tgtttttttt tggggtacag tctatgcctc gggcatccaa
     1921 gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac
     1981 gcagcagggc agtcgcccta aaacaaagtt aaacatcatg agggaagcgg tgatcgccga
     2041 agtatcgact caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt
     2101 gctggccgta catttgtacg gctccgcagt ggatggcggc ctgaagccac acagcgatat
     2161 tgatttgctg gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa
     2221 cgaccttttg gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt
     2281 caccattgtt gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca
     2341 atttggagaa tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga
     2401 cattgatctg gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc
     2461 agcggcggag gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga
     2521 aaccttaacg ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct
     2581 tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc
     2641 tgccgactgg gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag
     2701 acaggcttat cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga
     2761 atttgtccat tacgtaaaag gcgagatcac caaggtagtc ggcaaataac tgtcagacca
     2821 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     2881 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     2941 ctgagcgtca gaccccgtag aaaagatcaa aggatcttc
//
