LOCUS       sequence-459748-        2916 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 2916)
  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 2916)
  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-mKO2
FEATURES             Location/Qualifiers
     source          1..2916
                     /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             847..1503
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     primer_bind     complement(1543..1561)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1629..1733
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1955..2746
                     /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 tgagtgtgat taaaccagag atgaagatga ggtactacat ggacggctcc
      901 gtcaatgggc atgagttcac aattgaaggt gaaggcacag gcagacctta cgagggacat
      961 caagagatga cactacgcgt cacaatggcc gagggcgggc caatgccttt cgcgtttgac
     1021 ttagtgtcac acgtgttctg ttacggccac agagtattta ctaaatatcc agaagagata
     1081 ccagactatt tcaaacaagc atttcctgaa ggcctgtcat gggaaaggtc gttggagttc
     1141 gaagatggtg ggtccgcttc agtcagtgcg catataagcc ttagaggaaa caccttctac
     1201 cacaaatcca aatttactgg ggttaacttt cctgccgatg gtcctatcat gcaaaaccaa
     1261 agtgttgatt gggagccatc aaccgagaaa attactgcca gcgacggagt tctgaagggt
     1321 gatgttacga tgtacctaaa acttgaagga ggcggcaatc acaaatgcca aatgaagact
     1381 acttacaagg cggcaaaaga gattcttgaa atgccaggtg atcattacat cggccatcgc
     1441 ctcgtcagga aaaccgaagg caacattact gagcaggtag aagatgcagt agctcatagc
     1501 taagctttga gaccacgaag tggctcttca gtggacgaaa gggcctcgtg atacgcctat
     1561 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     1621 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1681 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgcgct
     1741 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
     1801 tcatggcttg ttatgactgt ttttttttgg ggtacagtct atgcctcggg catccaagca
     1861 gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca
     1921 gcagggcagt cgccctaaaa caaagttaaa catcatgagg gaagcggtga tcgccgaagt
     1981 atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct
     2041 ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gcgatattga
     2101 tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga
     2161 ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac
     2221 cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt
     2281 tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat
     2341 tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc
     2401 ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac
     2461 cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac
     2521 gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc
     2581 cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca
     2641 ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt
     2701 tgtccattac gtaaaaggcg agatcaccaa ggtagtcggc aaataactgt cagaccaagt
     2761 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2821 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2881 agcgtcagac cccgtagaaa agatcaaagg atcttc
//
