LOCUS       sequence-178368-        4665 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 4665)
  TITLE     Swint-Kruse lab plasmids
REFERENCE   2  (bases 1 to 4665)
  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..4665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..86
                     /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(106..125)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(1332..1349)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1332..1348)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1341..1363)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1516..1534)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1602..1706
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1707..1775,1776..2567)
                     /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(1925..1944)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2738..3326
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3227..3246
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3480..3497
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3512..3652
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3578..3597)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3738..3760
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3754..3945)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    4545..4566
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4581..4586,4587..4604,4605..4611)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     4641..4657
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4641..4657
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cgaattccga caccatcgaa tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg
       61 aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt
      121 atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg
      181 cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct caattacgcg ccatcagctc
      241 tggcgcgtag cctcaaactc aatcaaacac ataccattgg catgttgatc actgccagta
      301 ccaatccttt ctattcagaa ctggtgcgtg gcgttgaacg cagctgcttc gaacgcggtt
      361 atagtctcgt cctttgcaat accgaaggcg atgaacagcg gatgaatcgc aatctggaaa
      421 cgctgatgca aaaacgcgtt gatggcttgc tgttactgtg caccgaaacg catcaacctt
      481 cgcgtgaaat catgcaacgt tatccgacag tgcctactgt gatgatggac tgggctccgt
      541 tcgatggcga cagcgatctt attcaggata actcgttgct gggcggagac ttagcaacgc
      601 aatatctgat cgataaaggt catacccgta tcgcctgtat taccggcccg ctggataaaa
      661 ctccggcgcg cctgcggttg gaaggttatc gggcggcgat gaaacgtgcg ggtctcaaca
      721 ttcctgatgg ctatgaagtc actggtgatt ttgaatttaa cggcgggttt gacgctatgc
      781 gccaactgct atcacatccg ctgcgtccac aggccgtctt taccggaaat gacgctatgg
      841 ctgttggcgt ttaccaggcg ttatatcagg cagagttaca ggttccgcag gatatcgcgg
      901 tgattggcta tgacgatatc gaactggcaa gctttatgac gccaccatta accactatcc
      961 accaaccgaa agatgaactg ggggagctgg cgattgatgt actcatccat cggataaccc
     1021 agccgaccct tcagcaacaa cgattacaac ttactccgat tctgatggaa cgcggttcgg
     1081 cttagtaacc tcaggccgat actgtcgtcg tcccctcaaa ctggcagatg cacggttacg
     1141 atgcgcccat ctacaccaac gtgacctatc ccattaggtc aatccgccgt ttgttcccac
     1201 ggagaatccg acgggttgtt actcgctcac atttaatgtt gatgaaagct ggctacagga
     1261 aggccagacg cgaattattt ttgatggcgt tggaattccc ggggatccgt cgacctgcag
     1321 ccaagcttgg cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     1381 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     1441 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatgccg ttaacggatt
     1501 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     1561 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1621 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1681 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     1741 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     1801 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     1861 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     1921 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     1981 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2041 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2101 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2161 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2221 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     2281 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2341 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2401 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     2461 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2521 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     2581 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     2641 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     2701 gagcgtcagg ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2761 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2821 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2881 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2941 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3001 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3061 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3121 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3181 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3241 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3301 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3361 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3421 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3481 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     3541 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     3601 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggccg cgccccgaca
     3661 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     3721 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3781 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     3841 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     3901 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
     3961 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
     4021 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
     4081 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
     4141 tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat
     4201 aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac
     4261 cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc
     4321 gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct
     4381 caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc ccgccgttaa
     4441 cggccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     4501 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     4561 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggcgtg
     4621 gtactcataa caatttcaca caggaaacag ctatgaccat gatta
//
