LOCUS       sequence-459752-        2847 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insulator backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2847)
  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 2847)
  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 [backbone]
FEATURES             Location/Qualifiers
     source          1..2847
                     /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"
     protein_bind    904..925
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(940..945,946..963,964..970)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    978..994
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     983..1005
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1002..1018
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1002..1018
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1014..1337
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    1031..1087
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1088..1105)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1088..1104)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1097..1119)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1313..1332)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1474..1492)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1560..1664
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1886..2677
                     /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 ttgtcttcgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
      901 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
      961 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     1021 tgattacgcc aagcttgcat gcctgcaggt cgactctaga ggatccccgg gtaccgagct
     1081 cgaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     1141 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     1201 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt
     1261 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
     1321 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     1381 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgacgaag acaatacttg
     1441 agaccacgaa gtggctcttc agtggacgaa agggcctcgt gatacgccta tttttatagg
     1501 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     1561 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1621 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgcgc tcacgcaact
     1681 ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt
     1741 gttatgactg tttttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg
     1801 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag
     1861 tcgccctaaa acaaagttaa acatcatgag ggaagcggtg atcgccgaag tatcgactca
     1921 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
     1981 tttgtacggc tccgcagtgg atggcggcct gaagccacac agcgatattg atttgctggt
     2041 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
     2101 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
     2161 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
     2221 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
     2281 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
     2341 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
     2401 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
     2461 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
     2521 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
     2581 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccatta
     2641 cgtaaaaggc gagatcacca aggtagtcgg caaataactg tcagaccaag tttactcata
     2701 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2761 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2821 ccccgtagaa aagatcaaag gatcttc
//
