LOCUS       sequence-178371-        4613 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a chimeric LacI:TreR transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4613)
  TITLE     Swint-Kruse lab plasmids
REFERENCE   2  (bases 1 to 4613)
  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"
COMMENT     Sequence Label: LThT-pHG165
FEATURES             Location/Qualifiers
     source          1..4613
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        12..89
                     /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(109..128)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(1257..1273)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1257..1273)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1270..1292)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1281..1297
                     /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(1305..1311,1312..1329,1330..1335))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1350..1371
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1971..2162
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2156..2178)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2264..2404
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2319..2338
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2419..2436)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2590..3178)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2670..2689)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3349..4140,4141..4209))
                     /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     3972..3991
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4210..4314)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4382..4400
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     4553..4575
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4567..4584
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4568..4584
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ccgggaattc cgacaccatc gaatggtgca aaacctttcg cggtatggca tgatagcgcc
       61 cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag
      121 agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt
      181 ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gcatggattt tccccttccc
      241 gctctgcgcg cgctatgcgt gggcaaagcg ataaagtggt cgccatcatt gttacccgtc
      301 tggattcgtt gtcagaaaat ctcgccgttc aaaccatgct gccagcgttc tatgaacaag
      361 gttacgaccc aatcatgatg gaaagtcagt tttccccgca attagttgcc gaacatttgg
      421 gagtgctgaa acggcgtaat atcgacggcg tagtgctgtt cggttttact ggcataacag
      481 aagaaatgtt agcccactgg cagtcatcgc tggttctgct ggcgcgtgac gcaaaaggct
      541 ttgcttcggt ctgttatgac gacgaagggg caatcaaaat cctgatgcaa cggctgtatg
      601 accaggggca tcgtaatatc agttatctcg gcgtgccgca cagtgacgtg acaaccggta
      661 agcgacgtca cgaagcctac ctggcgttct gcaaagcgca taaactgcat cccgttgccg
      721 ccctgccagg gcttgctatg aagcaaggct atgagaacgt tgcaaaagtg attacgcctg
      781 aaactaccgc cttactgtgc gcaaccgaca cgctggcact tggcgcaagt aaatacctgc
      841 aagagcaacg catcgacacc ttgcaactgg cgagcgtcgg taatacgccg ttaatgaaat
      901 tcctccatcc ggagatcgta accgtagatc ccggttacgc cgaagctgga cgccaggcgg
      961 cttgccagtt gatcgcgcag gtaaccgggc gcagcgaacc gcaacaaatc atcatccccg
     1021 ccaccctgtc ctgatgacct gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc
     1081 acggttacga tgcgcccatc tacaccaacg tgacctatcc cattaggtca atccgccgtt
     1141 tgttcccacg gagaatccga cgggttgtta ctcgctcaca tttaatgttg atgaaagctg
     1201 gctacaggaa ggccagacgc gaattatttt tgatggcgtt ggaattcgta atcatggtca
     1261 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     1321 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     1381 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     1441 caacgcgcgg ggagaggcgg tttgcgtatt ggccgttaac ggcgggcagc gttgggtcct
     1501 ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt
     1561 gccttactgg ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct
     1621 gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa
     1681 gtctggaaac gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat
     1741 gctgctggct accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct
     1801 gagtgatttt tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt
     1861 ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca
     1921 tcggtatcat tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac
     1981 aggaaaaaac cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg
     2041 agaaactcaa cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc
     2101 acgctgatga gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     2161 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     2221 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     2281 cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact
     2341 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     2401 caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2461 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2521 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2581 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2641 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2701 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2761 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2821 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2881 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2941 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3001 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     3061 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3121 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3181 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3241 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3301 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3361 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3421 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3481 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3541 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3601 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3661 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3721 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3781 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3841 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3901 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3961 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4021 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4081 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4141 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4201 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4261 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4321 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     4381 aaataggcgt atcacgaggc cctttcgtct tcaagaatcc gttaacggca ttcgccattc
     4441 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     4501 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     4561 cgacgttgta aaacgacggc cagtgccaag cttggctgca ggtcgacgga tcc
//
