LOCUS       sequence-178208-        4699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a chimeric LacI:PurR transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4699)
  AUTHORS   Tungtur S, Meinhardt S, Swint-Kruse L
  TITLE     Comparing the functional roles of nonconserved sequence positions in
            homologous transcription repressors: implications for 
            sequence/function analyses.
  JOURNAL   J Mol Biol. 2010 Jan 29;395(4):785-802. doi: 
            10.1016/j.jmb.2009.10.001. Epub 2009 Oct 8.
  PUBMED    19818797
REFERENCE   2  (bases 1 to 4699)
  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: 
            "10.1016/j.jmb.2009.10.001"; journalName: "J Mol Biol"; date: 
            "2010-01-29- 29"; volume: "395"; issue: "4"; pages: "785-802"
COMMENT     Sequence Label: LPhP-pHG165
FEATURES             Location/Qualifiers
     source          1..4699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(559..1350,1351..1419))
                     /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     1182..1201
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1420..1524)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1592..1610
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     1763..1785
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1777..1794
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1778..1794
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1835..1912
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(1932..1951)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(3166..3182)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3166..3182)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3179..3201)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3190..3206
                     /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)."
     promoter        complement(join(3214..3220,3221..3238,3239..3244))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3259..3280
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             3880..4071
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4065..4087)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4173..4313
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4228..4247
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4328..4345)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4499..4699,1..388))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4579..4598)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
       61 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
      121 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
      181 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
      241 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
      301 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
      361 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
      421 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
      481 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
      541 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
      601 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
      661 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
      721 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
      781 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
      841 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
      901 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
      961 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     1021 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     1081 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     1141 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     1201 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     1261 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     1321 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     1381 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     1441 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     1501 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     1561 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     1621 tcaagaatcc gttaacggca ttcgccattc aggctgcgca actgttggga agggcgatcg
     1681 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     1741 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag
     1801 cttggctgca ggtcgacgga tccccgggaa ttccgacacc atcgaatggt gcaaaacctt
     1861 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
     1921 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
     1981 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
     2041 ggagctccac tactccccta gcgcggtggc gcgtagcctg gcggttaacc acaccaagtc
     2101 tatcggtttg ctggcgacca gcagcgaagc ggcctatttt gccgagatca ttgaagcagt
     2161 tgaaaaaaat tgcttccaga aaggttacac cctgattctg ggcaatgcgt ggaacaatct
     2221 tgagaaacag cgggcttatc tgtcgatgat ggcgcaaaaa cgcgtcgatg gtctgctggt
     2281 gatgtgttct gagtacccag agccgttgct ggcgatgctg gaagagtatc gccatatccc
     2341 aatggtggtc atggactggg gtgaagcaaa agctgacttc accgatgcgg tcattgataa
     2401 cgcgttcgaa ggcggctaca tggccgggcg ttatctgatt gaacgcggtc accgcgaaat
     2461 cggcgtcatc cccggcccgc tggaacgtaa caccggcgca ggccgccttg ccggttttat
     2521 gaaggcgatg gaagaagcga tgatcaaggt gccggaaagc tggattgtgc agggtgactt
     2581 tgaacctgaa tccggttatc gcgccatgca gcaaatcctg tcgcagccgc atcgccctac
     2641 tgccgtcttc tgtggtggcg atatcatggc aatgggcgca ctttgtgctg ctgatgaaat
     2701 gggcctgcgc gtcccgcagg atgtttcgct gatcggttat gataacgtgc gcaacgcgcg
     2761 ctattttacg ccggcgctga ccacgatcca tcagccaaaa gattcgctgg gtgaaacagc
     2821 gttcaacatg ctgttggatc gtatcgtcaa caaacgtgaa gaaccgcagt ctattgaagt
     2881 gcatccgcgc ttgattgaac gccgctccgt ggctgacggc ccgttccgcg actatcgtcg
     2941 ttaataacct gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc acggttacga
     3001 tgcgcccatc tacaccaacg tgacctatcc cattaggtca atccgccgtt tgttcccacg
     3061 gagaatccga cgggttgtta ctcgctcaca tttaatgttg atgaaagctg gctacaggaa
     3121 ggccagacgc gaattatttt tgatggcgtt gaattcgtaa tcatggtcat agctgtttcc
     3181 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3241 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3301 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3361 gagaggcggt ttgcgtattg gccgttaacg gcgggcagcg ttgggtcctg gccacgggtg
     3421 cgcatgatcg tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt
     3481 tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct
     3541 gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg
     3601 cggaagtcag cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta
     3661 ccctgtggaa cacctacatc tgtattaacg aagcgctggc attgaccctg agtgattttt
     3721 ctctggtccc gccgcatcca taccgccagt tgtttaccct cacaacgttc cagtaaccgg
     3781 gcatgttcat catcagtaac ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt
     3841 acccccatga acagaaatcc cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc
     3901 gcccttaaca tggcccgctt tatcagaagc cagacattaa cgcttctgga gaaactcaac
     3961 gagctggacg cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag
     4021 ctttaccgca gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     4081 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     4141 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     4201 agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     4261 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
     4321 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4381 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4441 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4501 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4561 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4621 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4681 tggcgctttc tcatagctc
//
