LOCUS       sequence-178400-        4700 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a chimeric LacI:GalR:PurR transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4700)
  AUTHORS   Meinhardt S, Manley MW Jr, Parente DJ, Swint-Kruse L
  TITLE     Rheostats and toggle switches for modulating protein function.
  JOURNAL   PLoS One. 2013 Dec 30;8(12):e83502. doi: 
            10.1371/journal.pone.0083502. eCollection 2013.
  PUBMED    24386217
REFERENCE   2  (bases 1 to 4700)
  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: "2013-12-30"; volume: "8(12):e83502. doi"; pages: " 
            10.1371/journal.pone.0083502. eCollection 2013"
COMMENT     Sequence Label: LGhP-pHG165
FEATURES             Location/Qualifiers
     source          1..4700
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        15..92
                     /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(112..131)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(1347..1363)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1347..1363)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1360..1382)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1371..1387
                     /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(1395..1401,1402..1419,1420..1425))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1440..1461
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2061..2252
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2246..2268)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2354..2494
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2409..2428
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2509..2526)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2680..3268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2760..2779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3439..4230,4231..4299))
                     /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     4062..4081
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4300..4404)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4472..4490
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     4643..4665
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4657..4674
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4658..4674
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tccccgggaa ttccgacacc atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc
       61 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg
      121 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg
      181 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctcagc tatcacccga
      241 acgccaacgc ccgtgcgctg gcgcagcaga ccactaaatc tatcggtttg ctggcgacca
      301 gcagcgaagc ggcctatttt gccgagatca ttgaagcagt tgaaaaaaat tgcttccaga
      361 aaggttacac cctgattctg ggcaatgcgt ggaacaatct tgagaaacag cgggcttatc
      421 tgtcgatgat ggcgcaaaaa cgcgtcgatg gtctgctggt gatgtgttct gagtacccag
      481 agccgttgct ggcgatgctg gaagagtatc gccatatccc aatggtggtc atggactggg
      541 gtgaagcaaa agctgacttc accgatgcgg tcattgataa cgcgttcgaa ggcggctaca
      601 tggccgggcg ttatctgatt gaacgcggtc accgcgaaat cggcgtcatc cccggcccgc
      661 tggaacgtaa caccggcgca ggccgccttg ccggttttat gaaggcgatg gaagaagcga
      721 tgatcaaggt gccggaaagc tggattgtgc agggtgactt tgaacctgaa tccggttatc
      781 gcgccatgca gcaaatcctg tcgcagccgc atcgccctac tgccgtcttc tgtggtggcg
      841 atatcatggc aatgggcgca ctttgtgctg ctgatgaaat gggcctgcgc gtcccgcagg
      901 atgtttcgct gatcggttat gataacgtgc gcaacgcgcg ctattttacg ccggcgctga
      961 ccacgatcca tcagccaaaa gattcgctgg gtgaaacagc gttcaacatg ctgttggatc
     1021 gtatcgtcaa caaacgtgaa gaaccgcagt ctattgaagt gcatccgcgc ttgattgaac
     1081 gccgctccgt ggctgacggc ccgttccgcg actatcgtcg ttaataacct gaggccgata
     1141 ctgtcgtcgt cccctcaaac tggcagatgc acggttacga tgcgcccatc tacaccaacg
     1201 tgacctatcc cattaggtca atccgccgtt tgttcccacg gagaatccga cgggttgtta
     1261 ctcgctcaca tttaatgttg atgaaagctg gctacaggaa ggccagacgc gaattatttt
     1321 tgatggcgtt ggaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1381 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1441 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1501 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1561 ggccgttaac ggcgggcagc gttgggtcct ggccacgggt gcgcatgatc gtgctcctgt
     1621 cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga
     1681 tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcaacaacat
     1741 gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca
     1801 ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat
     1861 ctgtattaac gaagcgctgg cattgaccct gagtgatttt tctctggtcc cgccgcatcc
     1921 ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa
     1981 cccgtatcgt gagcatcctc tctcgtttca tcggtatcat tacccccatg aacagaaatc
     2041 ccccttacac ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac atggcccgct
     2101 ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac gcggatgaac
     2161 aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc agctgcctcg
     2221 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     2281 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     2341 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct
     2401 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     2461 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
     2521 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2581 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2641 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2701 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2761 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2821 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2881 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2941 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3001 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3061 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3121 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3181 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3241 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3301 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3361 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3421 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3481 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3541 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3601 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3661 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3721 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3781 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3841 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3901 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3961 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4021 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4081 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4141 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4201 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4261 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4321 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4381 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     4441 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     4501 tcaagaatcc gttaacggca ttcgccattc aggctgcgca actgttggga agggcgatcg
     4561 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     4621 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag
     4681 cttggctgca ggtcgacgga
//
