LOCUS       sequence-174546-        5175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a chimeric LacI:RbsR transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5175)
  TITLE     Swint-Kruse lab plasmids
REFERENCE   2  (bases 1 to 5175)
  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: LRhR-pHG165a
FEATURES             Location/Qualifiers
     source          1..5175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(205..222)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(205..221)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      709..1222
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(796..815)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1006..1027
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    1638..1778
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1704..1723)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1864..1886
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1934..1952)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2020..2124
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2125..2193,2194..2985)
                     /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(2343..2362)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3155..3743
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3644..3663
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3897..3914
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3938..3959
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3974..3979,3980..3997,3998..4004)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4012..4028
                     /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     4017..4039
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4036..4052
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4036..4052
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4099..4176
                     /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(4196..4215)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tgaggccgat actgtcgtcg tcccctcaaa ctggcagatg cacggttacg atgcgcccat
       61 ctacaccaac gtaacctatc ccattacggt caatccgccg tttgttccca cggagaatcc
      121 gacgggttgt tactcgctca catttaatgt tgatgaaagc tggctacagg aaggccagac
      181 gcgaattatt tttgatggcg cggcactggc cgtcgtttta caacgtactg actgggaaaa
      241 ccctggcgtt acccaactta atcgccttgc agcacatccc cccttcgcca gctggcgtaa
      301 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgtagcctga atggcgaatg
      361 gtaattccag acgattgagc gtcaaaatgt aggtatttcc atgagcgttt ttcctgttgc
      421 aatggctggc ggtaatattg ttctggatat taccagcaag gccgatagtt tgagttcttc
      481 tactcaggca agtgatgtta ttactaatca aagaagtatt gcgacaacgg ttaatttgcg
      541 tgatggacag actcttttac tcggtggcct cactgattat aaaaacactt ctcaggattc
      601 tggcgtaccg ttcctgtcta aaatcccttt aatcggcctc ctgtttagct cccgctctga
      661 ttctaacgag gaaagcacgt tatacgtgct cgtcaaagca accatagtac gcgccctgta
      721 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
      781 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
      841 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
      901 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
      961 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     1021 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     1081 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     1141 acaaaatatt aacgtttaca atttaaatat ttgcttatac aatcttcctg tttttggggc
     1201 ttttctgatt atcaaccggg gtacatatga ttgacatgct agttttacga ttaccgttca
     1261 tcgattctct tgtttgctcc agactctcag gcaatgacct gatagccttt gtagagacct
     1321 ctcaaaaata gctaccctct ccggcatgaa tttatcagct agaacggttg aatatcatat
     1381 tgatggtgat ttgactgtct ccggcctttc tcacccgttt gaatctttac ctacacatta
     1441 ctcaggcatt gcatttaaaa tatatgaggg ttctaaaaat ttttatcctt gcgttgaaat
     1501 aaaggcttct cccgcaaaag tattacaggg tcataatgtt tttggtacaa ccgatttagc
     1561 tttatgctct gaggctttat tgcttaattt tgctaattct ttgccttgcc tgtatgattt
     1621 attggatgtt ggaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     1681 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     1741 atactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     1801 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     1861 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagt
     1921 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     1981 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2041 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2101 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     2161 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     2221 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2281 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2341 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     2401 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     2461 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     2521 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     2581 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     2641 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     2701 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     2761 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2821 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     2881 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2941 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     3001 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3061 aagatccttt ttgataatct catgaccaaa atccctaacg tgagttttcg ttccactgag
     3121 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3181 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3241 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3301 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3361 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3421 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3481 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctaccgc
     3541 gtgagcattg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3601 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3661 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3721 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3781 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3841 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3901 agtcagtgag cgaggaagcg gaagagcgca acgcaattaa tgtgagttag ctcactcatt
     3961 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     4021 gataacaatt tcacacagga aacagctatg accatgatta caagcttggc tcgaggtcga
     4081 cggatccccg ggaattccga caccatcgaa tggtgcaaaa cctttcgcgg tatggcatga
     4141 tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat
     4201 gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc
     4261 cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct caattacgcg
     4321 ccatcagctc tggcgcgtag cctcaaactc aatcaaacac ataccattgg catgttgatc
     4381 actgccagta ccaatccttt ctattcagaa ctggtgcgtg gcgttgaacg cagctgcttc
     4441 gaacgcggtt atagtctcgt cctttgcaat accgaaggcg atgaacagcg gatgaatcgc
     4501 aatctggaaa cgctgatgca aaaacgcgtt gatggcttgc tgttactgtg caccgaaacg
     4561 catcaacctt cgcgtgaaat catgcaacgt tatccgacag tgcctactgt gatgatggac
     4621 tgggctccgt tcgatggcga cagcgatctt attcaggata actcgttgct gggcggagac
     4681 ttagcaacgc aatatctgat cgataaaggt catacccgta tcgcctgtat taccggcccg
     4741 ctggataaaa ctccggcgcg cctgcggttg gaaggttatc gggcggcgat gaaacgtgcg
     4801 ggtctcaaca ttcctgatgg ctatgaagtc actggtgatt ttgaatttaa cggcgggttt
     4861 gacgctatgc gccaactgct atcacatccg ctgcgtcctc aggccgtctt taccggaaat
     4921 gacgctatgg ctgttggcgt ttaccaggcg ttatatcagg cagagttaca ggttccgcag
     4981 gatatcgcgg tgattggcta tgacgatatc gaactggcaa gctttatgac gccaccatta
     5041 accactatcc accaaccgaa agatgaactg ggggagctgg cgattgatgt actcatccat
     5101 cggataaccc agccgaccct tcagcaacaa cgattacaac ttactccgat tctgatggaa
     5161 cgcggttcgg cttag
//
