LOCUS       sequence-178291-        4637 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 4637)
  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 4637)
  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: LLhT-pHG165a V52A
FEATURES             Location/Qualifiers
     source          1..4637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    95..116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(131..136,137..154,155..161)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     217..233
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     217..233
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        250..327
                     /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(347..366)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(1519..1536)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1519..1535)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1528..1550)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1703..1721)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1789..1893
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1894..1962,1963..2754)
                     /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(2112..2131)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2925..3513
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3414..3433
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3667..3684
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3699..3839
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3765..3784)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3925..3947
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3941..4132)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
       61 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      121 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggcgtg gtactcataa
      181 caatttgtgt ggcgtggtac tcataacaat ttcacacagg aaacagctat gaccatgatt
      241 acgaattccg acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg
      301 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
      361 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
      421 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tcaattacat tcccaaccgc
      481 gccgcacaac aactggcggg caaacagagc aaagtggtcg ccatcattgt tacccgtctg
      541 gattcgttgt cagaaaatct cgccgttcaa accatgctgc cagcgttcta tgaacaaggt
      601 tacgacccaa tcatgatgga aagtcagttt tccccgcaat tagttgccga acatttggga
      661 gtgctgaaac ggcgtaatat cgacggcgta gtgctgttcg gttttactgg cataacagaa
      721 gaaatgttag cccactggca gtcatcgctg gttctgctgg cgcgtgacgc aaaaggcttt
      781 gcttcggtct gttatgacga cgaaggggca atcaaaatcc tgatgcaacg gctgtatgac
      841 caggggcatc gtaatatcag ttatctcggc gtgccgcaca gtgacgtgac aaccggtaag
      901 cgacgtcacg aagcctacct ggcgttctgc aaagcgcata aactgcatcc cgttgccgcc
      961 ctgccagggc ttgctatgaa gcaaggctat gagaacgttg caaaagtgat tacgcctgaa
     1021 actaccgcct tactgtgcgc aaccgacacg ctggcacttg gcgcaagtaa atacctgcaa
     1081 gagcaacgca tcgacacctt gcaactggcg agcgtcggta atacgccgtt aatgaaattc
     1141 ctccatccgg agatcgtaac cgtagatccc ggttacgccg aagctggacg ccaggcggct
     1201 tgccagttga tcgcgcaggt aaccgggcgc agcgaaccgc aacaaatcat catccccgcc
     1261 accctgtcct gatgacctga ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac
     1321 ggttacgatg cgcccatcta caccaacgtg acctatccca ttaggtcaat ccgccgtttg
     1381 ttcccacgga gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc
     1441 tacaggaagg ccagacgcga attatttttg atggcgttgg aattcccggg gatccgtcga
     1501 cctgcagcca agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     1561 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     1621 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgccgtta
     1681 acggattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     1741 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     1801 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1861 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     1921 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     1981 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2041 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2101 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     2161 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2221 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     2281 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     2341 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     2401 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2461 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2521 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2581 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     2641 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2701 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     2761 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     2821 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     2881 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2941 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3001 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3061 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3121 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3181 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3241 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3301 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3361 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3421 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3481 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3541 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3601 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3661 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     3721 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
     3781 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     3841 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     3901 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     3961 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     4021 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     4081 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     4141 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc
     4201 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac
     4261 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct
     4321 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc
     4381 ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac
     4441 cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg
     4501 ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc
     4561 gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ccaggaccca acgctgcccg
     4621 ccgttaacgg ccaatac
//
