LOCUS       sequence-177659-        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
FEATURES             Location/Qualifiers
     source          1..4704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    22..43
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(58..63,64..81,82..88)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     118..134
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     118..134
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        151..228
                     /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(248..267)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(1513..1530)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1513..1529)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1522..1544)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1697..1715)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1783..1887
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1888..1956,1957..2748)
                     /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(2106..2125)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2919..3507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3408..3427
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3661..3678
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3693..3833
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3759..3778)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3919..3941
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3935..4126)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
       61 acactttatg cttccggctc gtatgttgtg tggcgtggta ctcataacaa tttcacacag
      121 gaaacagcta tgaccatgat tacgaattcc gacaccatcg aatggtgcaa aacctttcgc
      181 ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta
      241 acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg
      301 aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag
      361 ctcaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagag cgaaacggtc
      421 ggtctggtcg ttggtgatgt ttccgatccg tttttcggtg caatggtgaa agcggtcgaa
      481 caggtggctt atcacaccgg taatttttta ttgattggca acggttacca caacgaacaa
      541 aaagagcgtc aggccattga gcaactgatc cgccatcgct gtgctgcgtt ggtcgtccat
      601 gccaaaatga tcccggatgc tgatttagcc tcattaatga aacaaatgcc cggtatggtg
      661 ctgatcaacc gtatcctgcc tggctttgaa aaccgttgta ttgctctgga cgatcgttac
      721 ggtgcctggc tggcaacgcg tcatttaatt cagcaaggtc atacccgcat tggttatctg
      781 tgctctaacc actctatttc tgacgccgaa gatcgtctgc aagggtatta cgatgccctt
      841 gctgaaagtg gtattgcggc caatgaccgg ctggtgacat ttggcgaacc agacgaaagc
      901 ggcggcgaac aggcaatgac caagcttttg ggacgaggaa gaaatttcac tgcggtagcc
      961 tgttataacg attcaatggc ggcgggtgcg atgggcgttc tcaatgataa tggtattgat
     1021 gtaccgggtg agatttcgtt aattggcttt gatgatgtgc tggtgtcacg ctatgtgcgt
     1081 ccgcgcctga ccaccgtgcg ttacccaatc gtgacgatgg cgacccaggc tgccgaactg
     1141 gctttggcgc tggcggataa tcgccctctc ccggaaatca ctaatgtctt tagtccgacg
     1201 ctggtacgtc gtcattcagt gtcaactccg tcgctggagg caagtcatca tgcaaccagc
     1261 gactaataac ctgaggccga tactgtcgtc gtcccctcaa actggcagat gcacggttac
     1321 gatgcgccca tctacaccaa cgtgacctat cccattaggt caatccgccg tttgttccca
     1381 cggagaatcc gacgggttgt tactcgctca catttaatgt tgatgaaagc tggctacagg
     1441 aaggccagac gcgaattatt tttgatggcg ttggaattcc cggggatccg tcgacctgca
     1501 gccaagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     1561 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     1621 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgcc gttaacggat
     1681 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     1741 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     1801 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1861 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     1921 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     1981 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     2041 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     2101 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     2161 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     2221 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     2281 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     2341 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     2401 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     2461 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     2521 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     2581 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     2641 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     2701 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     2761 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     2821 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2881 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2941 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3001 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3061 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3121 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3181 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3241 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3301 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3361 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3421 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3481 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3541 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     3601 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3661 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     3721 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     3781 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     3841 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3901 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3961 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     4021 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     4081 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     4141 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
     4201 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
     4261 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
     4321 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     4381 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     4441 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     4501 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     4561 tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgccgtta
     4621 acggccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     4681 cgacaggttt cccgactgga aagc
//
