LOCUS       sequence-459759-        2580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lvl 0 insulator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2580)
  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 2580)
  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-I-E1
FEATURES             Location/Qualifiers
     source          1..2580
                     /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"
     primer_bind     complement(1207..1225)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1293..1397
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1619..2410
                     /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 gtctcaggag
      841 gccacctttg gtcttgacat tcactaagga gtgggcctgg aataaaacca gaaatcccca
      901 ttctcagtct gccccaagcc ctccaattcc gagggcgctg tatgtataag ctcgggcagg
      961 caaaagtcga ctgtgagaac acgccagcag agggcgctgt ggccccatca gtccctgccc
     1021 actgaactct ccagaaggaa aagcggcaag gatgcaaaca agaaaatcag accagactgg
     1081 gagtctagac cctgcgtccc acgcggtccc accatggctt cctctttgga attttcaggg
     1141 ggacccagga agactgacac cgctgattac ttgagaccac gaagtggctc ttcagtggac
     1201 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     1261 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1321 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1381 attgaaaaag gaagagtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     1441 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt ttggggtaca
     1501 gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt
     1561 atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacatcat
     1621 gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga
     1681 gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg
     1741 cctgaagcca cacagcgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac
     1801 aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga
     1861 gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta
     1921 tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat
     1981 cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca
     2041 tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga
     2101 tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg
     2161 cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa
     2221 aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca
     2281 gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc
     2341 gcgcgcagat cagttggaag aatttgtcca ttacgtaaaa ggcgagatca ccaaggtagt
     2401 cggcaaataa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2461 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2521 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
