LOCUS       sequence-178174-        4637 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a chimeric LacI:TreR transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4637)
  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 4637)
  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: LLhT-pHG165a
FEATURES             Location/Qualifiers
     source          1..4637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      19..607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     508..527
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     761..778
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    793..933
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(859..878)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1019..1041
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1035..1226)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    1826..1847
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1862..1867,1868..1885,1886..1892)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     1948..1964
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1948..1964
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1981..2058
                     /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(2078..2097)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(3250..3267)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3250..3266)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3259..3281)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3434..3452)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3520..3624
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3625..3693,3694..4485)
                     /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(3843..3862)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
       61 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      121 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
      181 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      241 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      301 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      361 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      421 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      481 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      541 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      601 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      661 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
      721 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      781 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
      841 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
      901 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
      961 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     1021 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     1081 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     1141 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     1201 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     1261 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     1321 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     1381 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     1441 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     1501 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     1561 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     1621 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     1681 atgcgcaccc gtggccagga cccaacgctg cccgccgtta acggccaata cgcaaaccgc
     1741 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     1801 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     1861 ctttacactt tatgcttccg gctcgtatgt tgtgtggcgt ggtactcata acaatttgtg
     1921 tggcgtggta ctcataacaa tttcacacag gaaacagcta tgaccatgat tacgaattcc
     1981 gacaccatcg aatggtgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
     2041 caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt
     2101 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
     2161 cgggaaaaag tggaagcggc gatggcggag ctcaattaca ttcccaaccg cgtggcacaa
     2221 caactggcgg gcaaacagag caaagtggtc gccatcattg ttacccgtct ggattcgttg
     2281 tcagaaaatc tcgccgttca aaccatgctg ccagcgttct atgaacaagg ttacgaccca
     2341 atcatgatgg aaagtcagtt ttccccgcaa ttagttgccg aacatttggg agtgctgaaa
     2401 cggcgtaata tcgacggcgt agtgctgttc ggttttactg gcataacaga agaaatgtta
     2461 gcccactggc agtcatcgct ggttctgctg gcgcgtgacg caaaaggctt tgcttcggtc
     2521 tgttatgacg acgaaggggc aatcaaaatc ctgatgcaac ggctgtatga ccaggggcat
     2581 cgtaatatca gttatctcgg cgtgccgcac agtgacgtga caaccggtaa gcgacgtcac
     2641 gaagcctacc tggcgttctg caaagcgcat aaactgcatc ccgttgccgc cctgccaggg
     2701 cttgctatga agcaaggcta tgagaacgtt gcaaaagtga ttacgcctga aactaccgcc
     2761 ttactgtgcg caaccgacac gctggcactt ggcgcaagta aatacctgca agagcaacgc
     2821 atcgacacct tgcaactggc gagcgtcggt aatacgccgt taatgaaatt cctccatccg
     2881 gagatcgtaa ccgtagatcc cggttacgcc gaagctggac gccaggcggc ttgccagttg
     2941 atcgcgcagg taaccgggcg cagcgaaccg caacaaatca tcatccccgc caccctgtcc
     3001 tgatgacctg aggccgatac tgtcgtcgtc ccctcaaact ggcagatgca cggttacgat
     3061 gcgcccatct acaccaacgt gacctatccc attaggtcaa tccgccgttt gttcccacgg
     3121 agaatccgac gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag
     3181 gccagacgcg aattattttt gatggcgttg gaattcccgg ggatccgtcg acctgcagcc
     3241 aagcttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     3301 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     3361 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgccgtt aacggattct
     3421 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     3481 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3541 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3601 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3661 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3721 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3781 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     3841 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     3901 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3961 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     4021 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     4081 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     4141 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     4201 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     4261 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     4321 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     4381 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4441 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4501 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4561 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4621 gcgtcagacc ccgtaga
//
