LOCUS       sequence-178162-        4713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a chimeric LacI:GalS transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4713)
  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 4713)
  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: LLhS-pHG165a
FEATURES             Location/Qualifiers
     source          1..4713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     491..510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     744..761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    776..916
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(842..861)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1002..1024
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1018..1209)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    1809..1830
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1845..1850,1851..1868,1869..1875)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     1905..1921
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1905..1921
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1938..2015
                     /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(2035..2054)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(3309..3326)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3309..3325)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3318..3340)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3493..3511)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3579..3683
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3684..3752,3753..4544)
                     /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(3902..3921)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
       61 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
      121 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
      181 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
      241 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
      301 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
      361 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
      421 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
      481 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      541 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      601 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
      661 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
      721 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
      781 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
      841 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
      901 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
      961 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     1021 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     1081 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     1141 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     1201 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
     1261 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     1321 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
     1381 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca
     1441 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact
     1501 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca
     1561 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca
     1621 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggcca
     1681 ggacccaacg ctgcccgccg ttaacggcca atacgcaaac cgcctctccc cgcgcgttgg
     1741 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     1801 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     1861 ccggctcgta tgttgtgtgg cgtggtactc ataacaattt cacacaggaa acagctatga
     1921 ccatgattac gaattccgac accatcgaat ggtgcaaaac ctttcgcggt atggcatgat
     1981 agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg
     2041 tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc
     2101 acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctc aattacattc
     2161 ccaaccgcgt ggcacaacaa ctggcgggca aacagagcga caccattggc gtggtggtga
     2221 tggacgtttc tgatgcgttt ttcggcgcgc tggtaaaagc ggtggatctg gtcgctcagc
     2281 agcatcagaa atacgtgcta atcggcaata gctatcatga agcggaaaaa gagcgtcacg
     2341 ccattgaggt gttaattcgc cagcgttgta atgcgttgat tgttcactca aaagcattga
     2401 gtgacgatga actggcgcaa tttatggata acattcccgg tatggtgtta atcaaccgcg
     2461 ttgtgccggg gtacgcccat cgttgcgttt gcctggataa tctcagcggt gcccgaatgg
     2521 cgacgcgcat gttgctgaat aacggtcatc aacgtattgg ttatctttct tccagccacg
     2581 gcattgaaga tgacgccatg cgtaaagcag gctggatgag tgcgttgaaa gagcaggata
     2641 ttattccgcc ggaaagctgg attggcgctg gtacgccgga catgccgggc ggtgaggcgg
     2701 cgatggttaa actgctgggg cgcaatctac aacttaccgc tgtatttgct tataacgaca
     2761 atatggccgc tggcgcactg acagcattaa aagataatgg cattgcgatt ccgttacatc
     2821 tctcaatcat cgggttcgat gatattccca tcgcccgtta caccgacccg caattaacga
     2881 ccgtgcgtta tcccattgct tcaatggcga aattagccac cgaactggcc ttgcaggggg
     2941 cagcaggcaa tattgatcct cgtgccagcc actgttttat gccgacgtta gtgcgtcgcc
     3001 attctgtcgc aacacgccag aatgcggcgg cgatcactaa ctcaacaaat caggcgatgt
     3061 aataacctca ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg
     3121 cgcccatcta caccaacgtg acctatccca ttaggtcaat ccgccgtttg ttcccacgga
     3181 gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc tacaggaagg
     3241 ccagacgcga attatttttg atggcgttgg aattcccggg gatccgtcga cctgcagcca
     3301 agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     3361 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     3421 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgccgtta acggattctt
     3481 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     3541 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3601 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3661 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3721 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3781 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3841 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3901 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3961 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4021 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4081 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4141 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4201 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4261 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4321 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4381 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4441 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4501 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4561 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4621 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4681 cgtcagaccc cgtagaaaag atcaaaggat ctt
//
