LOCUS       sequence-71129-e        2951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains 8xLexA-binding elements.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2951)
  AUTHORS   Fujita T, Fujii H
  TITLE     Direct identification of insulator components by insertional 
            chromatin immunoprecipitation.
  JOURNAL   PLoS One. 2011;6(10):e26109. doi: 10.1371/journal.pone.0026109. Epub
            2011 Oct 17.
  PUBMED    22043306
REFERENCE   2  (bases 1 to 2951)
  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: "PLoS One.";
            date: "2011"; volume: "6(10):e26109. doi"; pages: " 
            10.1371/journal.pone.0026109. Epub 2011 Oct 17"
COMMENT     Sequence Label: 8xLexA-binding elements/pMD20
FEATURES             Location/Qualifiers
     source          1..2951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..40
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(341..358)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(341..357)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(350..372)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(565..584)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     684..706
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(744..762)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        830..934
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(935..1003,1004..1795)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1153..1172)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1966..2554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2455..2474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2708..2725
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2842..2863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2878..2883,2884..2901,2902..2908)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2916..2932
                     /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     2921..2943
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(2940..2951,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(2940..2951,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag ctatttaggt gacactatag gggaaagctt gcatgcctgc
       61 aggtcgactc tagaggatct actagtcata tggattaatc tagattctct atcgataggt
      121 acctcgactg ctgtatataa aaccagtggt tatatgtaca gtactgctgt atataaaacc
      181 agtggttata tgtacagtac gcctcgactg ctgtatataa aaccagtggt tatatgtaca
      241 gtactgctgt atataaaacc agtggttata tgtacagtac gctcgagatc cgctgaatag
      301 agtcgacaca atcggatccc cgggtaccga gctcgaattc actggccgtc gttttacaac
      361 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
      421 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
      481 gcctgaatgg cgaatggccc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
      541 acaccgcata tagtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
      601 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
      661 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
      721 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
      781 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
      841 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
      901 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
      961 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     1021 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     1081 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     1141 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     1201 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     1261 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     1321 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     1381 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     1441 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     1501 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     1561 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     1621 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     1681 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     1741 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     1801 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     1861 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     1921 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1981 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2041 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2101 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2161 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2221 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2281 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2341 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2401 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2461 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2521 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2581 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2641 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2701 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     2761 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     2821 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     2881 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     2941 aggaaacagc t
//
