LOCUS       sequence-178178-        4665 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a chimeric LacI:RbsR transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4665)
  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 4665)
  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: LLhR-pHG165a
FEATURES             Location/Qualifiers
     source          1..4665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     100..117
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    132..272
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(198..217)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     358..380
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(374..565)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    1165..1186
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1201..1206,1207..1224,1225..1231)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     1261..1277
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1261..1277
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1294..1371
                     /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(1391..1410)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(2617..2634)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2617..2633)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2626..2648)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2801..2819)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2887..2991
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2992..3060,3061..3852)
                     /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(3210..3229)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4023..4611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4512..4531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
       61 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
      121 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
      181 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
      241 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
      301 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
      361 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
      421 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
      481 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
      541 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
      601 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
      661 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
      721 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
      781 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
      841 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
      901 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
      961 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     1021 tgcgcacccg tggccaggac ccaacgctgc ccgccgttaa cggccaatac gcaaaccgcc
     1081 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     1141 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     1201 tttacacttt atgcttccgg ctcgtatgtt gtgtggcgtg gtactcataa caatttcaca
     1261 caggaaacag ctatgaccat gattacgaat tccgacacca tcgaatggtg caaaaccttt
     1321 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca
     1381 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg
     1441 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg
     1501 gagctcaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gagccatacc
     1561 attggcatgt tgatcactgc cagtaccaat cctttctatt cagaactggt gcgtggcgtt
     1621 gaacgcagct gcttcgaacg cggttatagt ctcgtccttt gcaataccga aggcgatgaa
     1681 cagcggatga atcgcaatct ggaaacgctg atgcaaaaac gcgttgatgg cttgctgtta
     1741 ctgtgcaccg aaacgcatca accttcgcgt gaaatcatgc aacgttatcc gacagtgcct
     1801 actgtgatga tggactgggc tccgttcgat ggcgacagcg atcttattca ggataactcg
     1861 ttgctgggcg gagacttagc aacgcaatat ctgatcgata aaggtcatac ccgtatcgcc
     1921 tgtattaccg gcccgctgga taaaactccg gcgcgcctgc ggttggaagg ttatcgggcg
     1981 gcgatgaaac gtgcgggtct caacattcct gatggctatg aagtcactgg tgattttgaa
     2041 tttaacggcg ggtttgacgc tatgcgccaa ctgctatcac atccgctgcg tcctcaggcc
     2101 gtctttaccg gaaatgacgc tatggctgtt ggcgtttacc aggcgttata tcaggcagag
     2161 ttacaggttc cgcaggatat cgcggtgatt ggctatgacg atatcgaact ggcaagcttt
     2221 atgacgccac cattaaccac tatccaccaa ccgaaagatg aactggggga gctggcgatt
     2281 gatgtactca tccatcggat aacccagccg acccttcagc aacaacgatt acaacttact
     2341 ccgattctga tggaacgcgg ttcggcttag taacctcagg ccgatactgt cgtcgtcccc
     2401 tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtgac ctatcccatt
     2461 aggtcaatcc gccgtttgtt cccacggaga atccgacggg ttgttactcg ctcacattta
     2521 atgttgatga aagctggcta caggaaggcc agacgcgaat tatttttgat ggcgttggaa
     2581 ttcccgggga tccgtcgacc tgcagccaag cttggcactg gccgtcgttt tacaacgtcg
     2641 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     2701 cagctggcgt aatagcgagg aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     2761 gaatggcgaa tgccgttaac ggattcttga agacgaaagg gcctcgtgat acgcctattt
     2821 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     2881 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2941 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3001 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3061 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3121 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3181 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3241 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3301 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3361 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3421 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3481 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3541 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3601 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3661 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3721 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3781 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3841 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3901 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3961 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4021 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4081 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4141 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     4201 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4261 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4321 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4381 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4441 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4501 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4561 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4621 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgct
//
