LOCUS       sequence-178200-        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-pHG165
FEATURES             Location/Qualifiers
     source          1..4682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(149..940,941..1009))
                     /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     772..791
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1010..1114)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1182..1200
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     1353..1375
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1367..1384
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1368..1384
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1425..1502
                     /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(1522..1541)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(2739..2755)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2739..2755)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2752..2774)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2763..2779
                     /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(2787..2793,2794..2811,2812..2817))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2832..2853
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             3453..3644
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3638..3660)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3746..3886
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3801..3820
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3901..3918)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4072..4660)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4152..4171)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
       61 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
      121 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
      181 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
      241 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
      301 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
      361 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
      421 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
      481 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
      541 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
      601 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
      661 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
      721 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
      781 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
      841 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
      901 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
      961 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     1021 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     1081 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     1141 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     1201 cctttcgtct tcaagaatcc gttaacggca ttcgccattc aggctgcgca actgttggga
     1261 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     1321 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     1381 cagtgccaag cttggctgca ggtcgacgga tccccgggaa ttccgacacc atcgaatggt
     1441 gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga
     1501 atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg
     1561 tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag
     1621 cggcgatggc ggagctcaat tacattccca accgcgtggc acaacaactg gcgggcaaac
     1681 agtcggacac cgtagccctg gtggtgtcgg aaaacaacca gaagctcttc gccgaaccct
     1741 tctatgccgg gatcgtgctc ggcgtggggg ttgctctgtc cgaacgggga ttccagttcg
     1801 tcctggccac gggccgctcc gggatagagc atgagcggct gggcggctac ctggccggac
     1861 agcacgtcga cggggtcctc ctgctgtcgc tccaccgcga cgacccgctg ccgcagatgc
     1921 tggacgaggc cggggtgccg tacgtctacg gcggccgtcc gctcggcgtc cccgaagaac
     1981 aggtgtccta tgtcgatatc gacaacatcg gcgggggacg ccaggccacc cagcggctga
     2041 tcgagaccgg gcaccggcgg atcgctacga tcgcgggccc gcaggacatg gtcgctggtg
     2101 tggaacgcct ccaggggtat cgcgaagcac tgctcgccgc ggggatggag tacgacgaga
     2161 cgctggtgag ctacggtgac ttcacctacg acagcggggt ggccgcgatg cgggagctgc
     2221 tggatcgggc ccccgacgtg gacgccgtgt tcgcggcctc cgacttgatg gggctggccg
     2281 cgctgcgggt gctgcgtgct tcgggacgcc gcgtgcccga ggatgtggcg gtggtcggct
     2341 acgacgactc gaccgtagcc gagcacgccg aaccgccgat gaccagcgtc aaccagccca
     2401 ccgagctgat gggccgggag atggcccggc tgctcgtgga ccggatcacc ggggagacca
     2461 ccgaaccggt gcggctggtg ctggagaccc atttgatggt gcgggaatcc gggtgataac
     2521 ctgaggccga tactgtcgtc gtcccctcaa actggcagat gcacggttac gatgcgccca
     2581 tctacaccaa cgtgacctat cccattaggt caatccgccg tttgttccca cggagaatcc
     2641 gacgggttgt tactcgctca catttaatgt tgatgaaagc tggctacagg aaggccagac
     2701 gcgaattatt tttgatggcg ttggaattcg taatcatggt catagctgtt tcctgtgtga
     2761 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2821 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2881 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2941 ggtttgcgta ttggccgtta acggcgggca gcgttgggtc ctggccacgg gtgcgcatga
     3001 tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga
     3061 atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct
     3121 gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt
     3181 cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg
     3241 gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt
     3301 cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt
     3361 catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca
     3421 tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta
     3481 acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg
     3541 acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc
     3601 gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     3661 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     3721 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
     3781 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     3841 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc
     3901 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3961 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4021 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4081 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4141 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4201 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4261 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4321 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4381 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4441 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4501 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4561 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4621 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4681 ta
//
