LOCUS       sequence-178292-        4704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a chimeric LacI:GalR transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4704)
  AUTHORS   Meinhardt S, Swint-Kruse L
  TITLE     Experimental identification of specificity determinants in the 
            domain linker of a LacI/GalR protein: bioinformatics-based 
            predictions generate true positives and false negatives.
  JOURNAL   Proteins. 2008 Dec;73(4):941-57. doi: 10.1002/prot.22121.
  PUBMED    18536016
REFERENCE   2  (bases 1 to 4704)
  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: "10.1002/prot"; 
            journalName: "Proteins"; date: "2008-12"; volume: "73"; issue: "4"; 
            pages: "941-57"
COMMENT     Sequence Label: LLhG-pHG165a E62K
FEATURES             Location/Qualifiers
     source          1..4704
                     /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(1353..1369)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1353..1369)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        complement(join(1399..1405,1406..1423,1424..1429))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1444..1465
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2065..2256
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2250..2272)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2358..2498
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2413..2432
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2513..2530)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2684..3272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2764..2783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3443..4234,4235..4303))
                     /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     4066..4085
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4304..4408)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4476..4494
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     4647..4669
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4661..4678
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4662..4678
                     /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 ggagctcaat tacattccca
      241 accgcgtggc acaacaactg gcgggcaaac agagcaaaac ggtcggtctg gtcgttggtg
      301 atgtttccga tccgtttttc ggtgcaatgg tgaaagcggt cgaacaggtg gcttatcaca
      361 ccggtaattt tttattgatt ggcaacggtt accacaacga acaaaaagag cgtcaggcca
      421 ttgagcaact gatccgccat cgctgtgctg cgttggtcgt ccatgccaaa atgatcccgg
      481 atgctgattt agcctcatta atgaaacaaa tgcccggtat ggtgctgatc aaccgtatcc
      541 tgcctggctt tgaaaaccgt tgtattgctc tggacgatcg ttacggtgcc tggctggcaa
      601 cgcgtcattt aattcagcaa ggtcataccc gcattggtta tctgtgctct aaccactcta
      661 tttctgacgc cgaagatcgt ctgcaagggt attacgatgc ccttgctgaa agtggtattg
      721 cggccaatga ccggctggtg acatttggcg aaccagacga aagcggcggc gaacaggcaa
      781 tgaccaagct tttgggacga ggaagaaatt tcactgcggt agcctgttat aacgattcaa
      841 tggcggcggg tgcgatgggc gttctcaatg ataatggtat tgatgtaccg ggtgagattt
      901 cgttaattgg ctttgatgat gtgctggtgt cacgctatgt gcgtccgcgc ctgaccaccg
      961 tgcgttaccc aatcgtgacg atggcgaccc aggctgccga actggctttg gcgctggcgg
     1021 ataatcgccc tctcccggaa atcactaatg tctttagtcc gacgctggta cgtcgtcatt
     1081 cagtgtcaac tccgtcgctg gaggcaagtc atcatgcaac cagcgactaa taacctgagg
     1141 ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca
     1201 ccaacgtgac ctatcccatt aggtcaatcc gccgtttgtt cccacggaga atccgacggg
     1261 ttgttactcg ctcacattta atgttgatga aagctggcta caggaaggcc agacgcgaat
     1321 tatttttgat ggcgttggaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     1381 tatgagtacc acgccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     1441 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1501 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     1561 tattggccgt taacggcggg cagcgttggg tcctggccac gggtgcgcat gatcgtgctc
     1621 ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca
     1681 ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca
     1741 acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc
     1801 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct
     1861 acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc
     1921 atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca
     1981 gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga
     2041 aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc
     2101 cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat
     2161 gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc
     2221 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     2281 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     2341 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
     2401 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     2461 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
     2521 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2581 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2641 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2701 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2761 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2821 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2881 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2941 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3001 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3061 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3121 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3181 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3241 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3301 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3361 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3421 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3481 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3541 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3601 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3661 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3721 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3781 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3841 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3901 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3961 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4021 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4081 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4141 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4201 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4261 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4321 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4381 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     4441 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     4501 gtcttcaaga atccgttaac ggcattcgcc attcaggctg cgcaactgtt gggaagggcg
     4561 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     4621 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
     4681 caagcttggc tgcaggtcga cgga
//
