LOCUS       sequence-178393-        4682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a chimeric LacI:CelR transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4682)
  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 4682)
  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: LLhE_AW-pHG165
FEATURES             Location/Qualifiers
     source          1..4682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        19..96
                     /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(116..135)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(1333..1349)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1333..1349)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1346..1368)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1357..1373
                     /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)."
     promoter        complement(join(1381..1387,1388..1405,1406..1411))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1426..1447
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2047..2238
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2232..2254)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2340..2480
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2395..2414
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2495..2512)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2666..3254)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2746..2765)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3425..4216,4217..4285))
                     /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     4048..4067
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4286..4390)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4458..4476
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     4629..4651
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4643..4660
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4644..4660
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cggatccccg ggaattccga caccatcgaa tggtgcaaaa cctttcgcgg tatggcatga
       61 tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat
      121 gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc
      181 cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct caattacgta
      241 cccaaccgcg tggcacaagc gctggcgggc aaacgatcgg acaccgtagc cctggtggtg
      301 tcggaaaaca accagaagct cttcgccgaa cccttctatg ccgggatcgt gctcggcgtg
      361 ggggttgctc tgtccgaacg gggattccag ttcgtcctgg ccacgggccg ctccgggata
      421 gagcatgagc ggctgggcgg ctacctggcc ggacagcacg tcgacggggt cctcctgctg
      481 tcgctccacc gcgacgaccc gctgccgcag atgctggacg aggccggggt gccgtacgtc
      541 tacggcggcc gtccgctcgg cgtccccgaa gaacaggtgt cctatgtcga tatcgacaac
      601 atcggcgggg gacgccaggc cacccagcgg ctgatcgaga ccgggcaccg gcggatcgct
      661 acgatcgcgg gcccgcagga catggtcgct ggtgtggaac gcctccaggg gtatcgcgaa
      721 gcactgctcg ccgcggggat ggagtacgac gagacgctgg tgagctacgg tgacttcacc
      781 tacgacagcg gggtggccgc gatgcgggag ctgctggatc gggcccccga cgtggacgcc
      841 gtgttcgcgg cctccgactt gatggggctg gccgcgctgc gggtgctgcg tgcttcggga
      901 cgccgcgtgc ccgaggatgt ggcggtggtc ggctacgacg actcgaccgt agccgagcac
      961 gccgaaccgc cgatgaccag cgtcaaccag cccaccgagc tgatgggccg ggagatggcc
     1021 cggctgctcg tggaccggat caccggggag accaccgaac cggtgcggct ggtgctggag
     1081 acccatttga tggtgcggga atccgggtga taacctgagg ccgatactgt cgtcgtcccc
     1141 tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtgac ctatcccatt
     1201 aggtcaatcc gccgtttgtt cccacggaga atccgacggg ttgttactcg ctcacattta
     1261 atgttgatga aagctggcta caggaaggcc agacgcgaat tatttttgat ggcgttggaa
     1321 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     1381 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     1441 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     1501 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattggcc gttaacggcg
     1561 ggcagcgttg ggtcctggcc acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg
     1621 ctaggctggc ggggttgcct tactggttag cagaatgaat caccgatacg cgagcgaacg
     1681 tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat ggtcttcggt
     1741 ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg
     1801 atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt attaacgaag
     1861 cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac cgccagttgt
     1921 ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg tatcgtgagc
     1981 atcctctctc gtttcatcgg tatcattacc cccatgaaca gaaatccccc ttacacggag
     2041 gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat cagaagccag
     2101 acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc agacatctgt
     2161 gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg tttcggtgat
     2221 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     2281 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     2341 gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat
     2401 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     2461 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2521 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2581 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2641 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2701 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2761 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2821 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2881 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2941 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3001 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3061 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     3121 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3181 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3241 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3301 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3361 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3421 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3481 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3541 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3601 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     3661 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     3721 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     3781 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     3841 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     3901 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     3961 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4021 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4081 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4141 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4201 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4261 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4321 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4381 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     4441 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaatccgtta
     4501 acggcattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     4561 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     4621 agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagcttg gctgcaggtc
     4681 ga
//
