LOCUS       sequence-178396-        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   Sousa FL, Parente DJ, Shis DL, Hessman JA, Chazelle A, Bennett MR, 
            Teichmann SA, Swint-Kruse L
  TITLE     AlloRep: A Repository of Sequence, Structural and Mutagenesis Data 
            for the LacI/GalR Transcription Regulators.
  JOURNAL   J Mol Biol. 2016 Feb 22;428(4):671-8. doi: 
            10.1016/j.jmb.2015.09.015. Epub 2015 Sep 25.
  PUBMED    26410588
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: 
            "10.1016/j.jmb.2015.09.015"; journalName: "J Mol Biol"; date: 
            "2016-02-22- 22"; volume: "428"; issue: "4"; pages: "671-8"
COMMENT     Sequence Label: LLhE_AW-pHG165 H118R
FEATURES             Location/Qualifiers
     source          1..4682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..83
                     /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(103..122)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(1320..1336)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1320..1336)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1333..1355)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1344..1360
                     /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(1368..1374,1375..1392,1393..1398))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1413..1434
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2034..2225
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2219..2241)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2327..2467
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2382..2401
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2482..2499)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2653..3241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2733..2752)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3412..4203,4204..4272))
                     /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     4035..4054
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4273..4377)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4445..4463
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     4616..4638
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4630..4647
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4631..4647
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 attccgacac catcgaatgg tgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
       61 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
      121 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
      181 aaacgcggga aaaagtggaa gcggcgatgg cggagctcaa ttacgtaccc aaccgcgtgg
      241 cacaagcgct ggcgggcaaa cgatcggaca ccgtagccct ggtggtgtcg gaaaacaacc
      301 agaagctctt cgccgaaccc ttctatgccg ggatcgtgct cggcgtgggg gttgctctgt
      361 ccgaacgggg attccagttc gtcctggcca cgggccgctc cgggatagag catgagcggc
      421 tgggcggcta cctggccgga cagcgcgtcg acggggtcct cctgctgtcg ctccaccgcg
      481 acgacccgct gccgcagatg ctggacgagg ccggggtgcc gtacgtctac ggcggccgtc
      541 cgctcggcgt ccccgaagaa caggtgtcct atgtcgatat cgacaacatc ggcgggggac
      601 gccaggccac ccagcggctg atcgagaccg ggcaccggcg gatcgctacg atcgcgggcc
      661 cgcaggacat ggtcgctggt gtggaacgcc tccaggggta tcgcgaagca ctgctcgccg
      721 cggggatgga gtacgacgag acgctggtga gctacggtga cttcacctac gacagcgggg
      781 tggccgcgat gcgggagctg ctggatcggg cccccgacgt ggacgccgtg ttcgcggcct
      841 ccgacttgat ggggctggcc gcgctgcggg tgctgcgtgc ttcgggacgc cgcgtgcccg
      901 aggatgtggc ggtggtcggc tacgacgact cgaccgtagc cgagcacgcc gaaccgccga
      961 tgaccagcgt caaccagccc accgagctga tgggccggga gatggcccgg ctgctcgtgg
     1021 accggatcac cggggagacc accgaaccgg tgcggctggt gctggagacc catttgatgg
     1081 tgcgggaatc cgggtgataa cctgaggccg atactgtcgt cgtcccctca aactggcaga
     1141 tgcacggtta cgatgcgccc atctacacca acgtgaccta tcccattagg tcaatccgcc
     1201 gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag
     1261 ctggctacag gaaggccaga cgcgaattat ttttgatggc gttggaattc gtaatcatgg
     1321 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1381 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1441 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     1501 ggccaacgcg cggggagagg cggtttgcgt attggccgtt aacggcgggc agcgttgggt
     1561 cctggccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg
     1621 gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct
     1681 gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt
     1741 aaagtctgga aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag
     1801 gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac
     1861 cctgagtgat ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac
     1921 gttccagtaa ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt
     1981 tcatcggtat cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca
     2041 aacaggaaaa aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc
     2101 tggagaaact caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg
     2161 accacgctga tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     2221 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     2281 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     2341 agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt
     2401 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     2461 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2521 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2581 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2641 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2701 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2761 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2821 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2881 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2941 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3001 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3061 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     3121 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3181 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3241 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3301 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3361 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3421 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3481 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3541 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3601 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3661 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3721 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3781 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3841 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3901 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3961 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4021 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4081 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4141 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4201 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4261 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4321 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4381 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     4441 taaaaatagg cgtatcacga ggccctttcg tcttcaagaa tccgttaacg gcattcgcca
     4501 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     4561 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     4621 tcacgacgtt gtaaaacgac ggccagtgcc aagcttggct gcaggtcgac ggatccccgg
     4681 ga
//
