LOCUS       sequence-178367-        4676 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Constitutive expression of a chimeric LacI:CytR transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4676)
  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 4676)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: LLhC-pHG165a Q55I
FEATURES             Location/Qualifiers
     source          1..4676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..87
                     /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(107..126)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(1342..1359)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1342..1358)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1351..1373)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1526..1544)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1612..1716
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1717..1785,1786..2577)
                     /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(1935..1954)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2748..3336
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3237..3256
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3490..3507
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3522..3662
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3588..3607)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3748..3770
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3764..3955)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    4555..4576
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4591..4596,4597..4614,4615..4621)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4629..4645
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4634..4656
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4653..4669
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4653..4669
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 acgaattccg acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg
       61 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
      121 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
      181 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tcaattacat tcccaatcgc
      241 gtggcacaaa tcctggcggg caaacagagc cgcaccattc tggtgattgt cccggatatc
      301 tgcgatccct tctttagcga aattattcgc ggtatcgaag ttacggcggc aaatcacgga
      361 tatctggtgc tgattggcga ctgtgcgcat caaaatcagc aggaaaaaac ctttatcgat
      421 ttgatcatca ccaagcaaat tgatggcatg ttgttgctgg gttcaaggct gccgtttgat
      481 gccagcattg aggaacagcg taatctgccg ccgatggtga tggcgaacga atttgcaccg
      541 gagctggagc tgcctacagt tcatatcgac aatctgaccg ccgcatttga tgcagtaaat
      601 tatttatatg agcaagggca taaacggatt ggctgtatag ccggtcccga agagatgccg
      661 ctgtgtcact accgcctgca aggctatgtt caggcgctgc gtcgctgcgg cattatggtt
      721 gatccgcaat acatcgcccg tggcgacttc accttcgaag ccggaagcaa agcgatgcag
      781 cagctgcttg atctcccaca accgcctact gctgtcttct gccatagcga tgtgatggcg
      841 ctcggcgcac tttctcaggc aaaacgccag gggctgaaag tcccggaaga cctttccata
      901 atcggttttg ataacatcga cctgacgcaa ttttgtgatc cgccgctgac aaccatcgcg
      961 cagccgcgtt acgaaatcgg tcgggaagct atgctgttat tgcttgatca aatgcagggg
     1021 caacacgttg gcagtggctc tcgtttaatg gactgcgaac ttatcatccg gggatcaaca
     1081 cgcgcgttac cttaataacc tgaggccgat actgtcgtcg tcccctcaaa ctggcagatg
     1141 cacggttacg atgcgcccat ctacaccaac gtgacctatc ccattaggtc aatccgccgt
     1201 ttgttcccac ggagaatccg acgggttgtt actcgctcac atttaatgtt gatgaaagct
     1261 ggctacagga aggccagacg cgaattattt ttgatggcgt tggaattccc ggggatccgt
     1321 cgacctgcag ccaagcttgg cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     1381 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     1441 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatgccg
     1501 ttaacggatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     1561 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     1621 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     1681 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     1741 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     1801 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     1861 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     1921 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     1981 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2041 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2101 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2161 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2221 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     2281 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2341 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2401 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2461 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2521 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     2581 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     2641 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     2701 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2761 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2821 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2881 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2941 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3001 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3061 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3121 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3181 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3241 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3301 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3361 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3421 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3481 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     3541 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     3601 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     3661 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3721 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3781 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     3841 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     3901 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     3961 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     4021 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     4081 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     4141 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     4201 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     4261 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     4321 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     4381 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc
     4441 ccgccgttaa cggccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     4501 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     4561 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     4621 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgatt
//
