LOCUS       sequence-178170-        5094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a chimeric LacI:FruR transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5094)
  AUTHORS   Meinhardt S, Manley MW Jr, Becker NA, Hessman JA, Maher LJ 3rd, 
            Swint-Kruse L
  TITLE     Novel insights from hybrid LacI/GalR proteins: family-wide 
            functional attributes and biologically significant variation in 
            transcription repression.
  JOURNAL   Nucleic Acids Res. 2012 Nov;40(21):11139-54. doi: 
            10.1093/nar/gks806. Epub 2012 Sep 10.
  PUBMED    22965134
REFERENCE   2  (bases 1 to 5094)
  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/gks806"; journalName: "Nucleic Acids Res"; date: 
            "2012-11"; volume: "40"; issue: "21"; pages: "11139-54"
COMMENT     Sequence Label: LLhF-pHG165a
FEATURES             Location/Qualifiers
     source          1..5094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      97..685
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     586..605
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     839..856
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    880..901
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(916..921,922..939,940..946)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    954..970
                     /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     959..981
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     978..994
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     978..994
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1044..1121
                     /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(1141..1160)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(2340..2357)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2340..2356)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2349..2371)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2797..3310
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2884..2903)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3094..3115
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3790..3809)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3909..3931
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3969..3987)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4055..4159
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4160..4228,4229..5020)
                     /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(4378..4397)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
       61 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
      121 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
      181 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
      241 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
      301 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
      361 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
      421 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
      481 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
      541 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
      601 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
      661 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
      721 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
      781 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
      841 cgagtcagtg agcgaggaag cggaagagcg caacgcaatt aatgtgagtt agctcactca
      901 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      961 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc ttggctcgag
     1021 gtcgacggat ccccgggaat tccgacacca tcgaatggtg caaaaccttt cgcggtatgg
     1081 catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
     1141 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
     1201 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctcaatt
     1261 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gagccgttct attggtcttg
     1321 tgatccccga tctggagaac accagctata cccgcatcgc taactatctt gaacgccagg
     1381 cgcggcaacg gggttatcaa ctgctgattg cctgctcaga agatcagcca gacaacgaaa
     1441 tgcggtgcat tgagcacctt ttacagcgtc aggttgatgc cattattgtt tcgacgtcgt
     1501 tgcctcctga gcatcctttt tatcaacgct gggctaacga cccgttcccg attgtcgcgc
     1561 tggaccgcgc cctcgatcgt gaacacttca ccagcgtggt tggtgccgat caggatgatg
     1621 ccgaaatgct ggcggaagag ttacgtaagt ttcccgccga gacggtgctt tatcttggtg
     1681 cgctaccgga gctttctgtc agcttcctgc gtgaacaagg tttccgtact gcctggaaag
     1741 atgatccgcg cgaagtgcat ttcctgtatg ccaacagcta tgagcgggag gcggctgccc
     1801 agttattcga aaaatggctg gaaacgcatc cgatgccgca ggcgctgttc acaacgtcgt
     1861 ttgcgttgtt gcaaggagtg atggatgtca cgctgcgtcg cgacggcaaa ctgccttctg
     1921 acctggcaat tgccaccttt ggcgataacg aactgctcga cttcttacag tgtccggtgc
     1981 tggcagtggc tcaacgtcac cgcgatgtcg cagagcgtgt gctggagatt gtcctggcaa
     2041 gcctggacga accgcgtaag ccaaaacctg gtttaacgcg cattaaacgt aatctctatc
     2101 gccgcggcgt gctcagccgt agctaataac ctgaggccga tactgtcgtc gtcccctcaa
     2161 actggcagat gcacggttac gatgcgccca tctacaccaa cgtgacctat cccattaggt
     2221 caatccgccg tttgttccca cggagaatcc gacgggttgt tactcgctca catttaatgt
     2281 tgatgaaagc tggctacagg aaggccagac gcgaattatt tttgatggcg ttggaattca
     2341 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     2401 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     2461 ccttcccaac agttgcgcag cctgaatggc gaatggcgat tccgttgcaa tggctggcgg
     2521 taatattgtt ctggatatta ccagcaaggc cgatagtttg agttcttcta ctcaggcaag
     2581 tgatgttatt actaatcaaa gaagtattgc gacaacggtt aatttgcgtg atggacagac
     2641 tcttttactc ggtggcctca ctgattataa aaacacttct caggattctg gcgtaccgtt
     2701 cctgtctaaa atccctttaa tcggcctcct gtttagctcc cgctctgatt ctaacgagga
     2761 aagcacgtta tacgtgctcg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     2821 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     2881 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2941 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3001 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3061 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3121 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     3181 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     3241 cgcttacaat ttaaatattt gcttatacaa tcttcctgtt tttggggctt ttctgattat
     3301 caaccggggt acatatgatt gacatgctag ttttacgatt accgttcatc gattctcttg
     3361 tttgctccag actctcaggc aatgacctga tagcctttgt agacctctca aaaatagcta
     3421 ccctctccgg catgaattta tcagctagaa cggttgaata tcatattgat ggtgatttga
     3481 ctgtctccgg cctttctcac ccgtttgaat ctttacctac acattactca ggcattgcat
     3541 ttaaaatata tgagggttct aaaaattttt atccttgcgt tgaaataaag gcttctcccg
     3601 caaaagtatt acagggtcat aatgtttttg gtacaaccga tttagcttta tgctctgagg
     3661 ctttattgct taattttgct aattctttgc cttgcctgta tgatttattg gatgttggaa
     3721 tcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     3781 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     3841 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3901 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     3961 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     4021 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4081 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4141 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4201 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4261 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4321 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4381 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4441 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4501 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4561 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4621 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4681 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4741 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4801 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4861 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4921 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4981 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5041 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggt
//
