LOCUS       sequence-178370-        4673 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a chimeric LacI:FruR transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4673)
  TITLE     Swint-Kruse lab plasmids
REFERENCE   2  (bases 1 to 4673)
  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: LFhF-pHG165
FEATURES             Location/Qualifiers
     source          1..4673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        15..92
                     /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(112..131)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(1344..1361)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1344..1360)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1353..1375)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1528..1546)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1614..1718
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1719..1787,1788..2579)
                     /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(1937..1956)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2750..3338
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3239..3258
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3492..3509
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3524..3664
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3590..3609)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3750..3772
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3766..3957)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    4557..4578
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4593..4598,4599..4616,4617..4623)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4631..4647
                     /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)."
     primer_bind     4636..4658
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4655..4671
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4655..4671
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tgattacgaa ttccgacacc atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc
       61 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg
      121 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg
      181 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagcacaat taccacccga
      241 acgccgtggc agctgggctt cgtgctggac gcacacgttc tattggtctt gtgatccccg
      301 atctggagaa caccagctat acccgcatcg ctaactatct tgaacgccag gcgcggcaac
      361 ggggttatca actgctgatt gcctgctcag aagatcagcc agacaacgaa atgcggtgca
      421 ttgagcacct tttacagcgt caggttgatg ccattattgt ttcgacgtcg ttgcctcctg
      481 agcatccttt ttatcaacgc tgggctaacg acccgttccc gattgtcgcg ctggaccgcg
      541 ccctcgatcg tgaacacttc accagcgtgg ttggtgccga tcaggatgat gccgaaatgc
      601 tggcggaaga gttacgtaag tttcccgccg agacggtgct ttatcttggt gcgctaccgg
      661 agctttctgt cagcttcctg cgtgaacaag gtttccgtac tgcctggaaa gatgatccgc
      721 gcgaagtgca tttcctgtat gccaacagct atgagcggga ggcggctgcc cagttattcg
      781 aaaaatggct ggaaacgcat ccgatgccgc aggcgctgtt cacaacgtcg tttgcgttgt
      841 tgcaaggagt gatggatgtc acgctgcgtc gcgacggcaa actgccttct gacctggcaa
      901 ttgccacctt tggcgataac gaactgctcg acttcttaca gtgtccggtg ctggcagtgg
      961 ctcaacgtca ccgcgatgtc gcagagcgtg tgctggagat tgtcctggca agcctggacg
     1021 aaccgcgtaa gccaaaacct ggtttaacgc gcattaaacg taatctctat cgccgcggcg
     1081 tgctcagccg tagctaataa cctgaggccg atactgtcgt cgtcccctca aactggcaga
     1141 tgcacggtta cgatgcgccc atctacacca acgtgaccta tcccattagg tcaatccgcc
     1201 gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag
     1261 ctggctacag gaaggccaga cgcgaattat ttttgatggc gttggaattc ccggggatcc
     1321 gtcgacctgc agccaagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     1381 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     1441 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgc
     1501 cgttaacgga ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     1561 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     1621 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1681 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     1741 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     1801 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     1861 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     1921 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     1981 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     2041 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     2101 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     2161 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     2221 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     2281 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     2341 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     2401 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     2461 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     2521 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     2581 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     2641 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     2701 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2761 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2821 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2881 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2941 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3001 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3061 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3121 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3181 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3241 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3301 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3361 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3421 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3481 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     3541 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct
     3601 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     3661 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3721 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3781 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     3841 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     3901 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
     3961 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
     4021 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     4081 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     4141 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
     4201 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
     4261 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
     4321 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
     4381 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct
     4441 gcccgccgtt aacggccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     4501 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     4561 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     4621 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga cca
//
