LOCUS       sequence-178285-        4706 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Constitutive expression of a chimeric LacI:PurR:GalR transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4706)
  TITLE     Swint-Kruse lab plasmids
REFERENCE   2  (bases 1 to 4706)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: LPhG-pHG165
FEATURES             Location/Qualifiers
     source          1..4706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..85
                     /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(105..124)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(1370..1387)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1370..1386)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1379..1401)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1554..1572)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1640..1744
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1745..1813,1814..2605)
                     /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(1963..1982)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2776..3364
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3265..3284
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3518..3535
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3550..3690
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3616..3635)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3776..3798
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3792..3983)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    4583..4604
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4619..4624,4625..4642,4643..4649)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4657..4673
                     /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     4662..4684
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4681..4697
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4681..4697
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gaattccgac accatcgaat ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga
       61 agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta
      121 tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc
      181 gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctc cactactccc ctagcgcggt
      241 ggcgcgtagc ctgaaggtta accacaccga aacggtcggt ctggtcgttg gtgatgtttc
      301 cgatccgttt ttcggtgcaa tggtgaaagc ggtcgaacag gtggcttatc acaccggtaa
      361 ttttttattg attggcaacg gttaccacaa cgaacaaaaa gagcgtcagg ccattgagca
      421 actgatccgc catcgctgtg ctgcgttggt cgtccatgcc aaaatgatcc cggatgctga
      481 tttagcctca ttaatgaaac aaatgcccgg tatggtgctg atcaaccgta tcctgcctgg
      541 ctttgaaaac cgttgtattg ctctggacga tcgttacggt gcctggctgg caacgcgtca
      601 tttaattcag caaggtcata cccgcattgg ttatctgtgc tctaaccact ctatttctga
      661 cgccgaagat cgtctgcaag ggtattacga tgcccttgct gaaagtggta ttgcggccaa
      721 tgaccggctg gtgacatttg gcgaaccaga cgaaagcggc ggcgaacagg caatgaccaa
      781 gcttttggga cgaggaagaa atttcactgc ggtagcctgt tataacgatt caatggcggc
      841 gggtgcgatg ggcgttctca atgataatgg tattgatgta ccgggtgaga tttcgttaat
      901 tggctttgat gatgtgctgg tgtcacgcta tgtgcgtccg cgcctgacca ccgtgcgtta
      961 cccaatcgtg acgatggcga cccaggctgc cgaactggct ttggcgctgg cggataatcg
     1021 ccctctcccg gaaatcacta atgtctttag tccgacgctg gtacgtcgtc attcagtgtc
     1081 aactccgtcg ctggaggcaa gtcatcatgc aaccagcgac taataacctg aggccgatac
     1141 tgtcgtcgtc ccctcaaact ggcagatgca cggttacgat gcgcccatct acaccaacgt
     1201 gacctatccc attaggtcaa tccgccgttt gttcccacgg agaatccgac gggttgttac
     1261 tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg aattattttt
     1321 gatggcgttg gaattcccgg ggatccgtcg acctgcagcc aagcttggca ctggccgtcg
     1381 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     1441 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     1501 agttgcgcag cctgaatggc gaatgccgtt aacggattct tgaagacgaa agggcctcgt
     1561 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     1621 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     1681 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     1741 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     1801 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttagg
     1861 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     1921 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     1981 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2041 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     2101 gttatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     2161 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     2221 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     2281 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     2341 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     2401 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     2461 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     2521 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     2581 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     2641 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     2701 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2761 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2821 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2881 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     2941 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3001 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3061 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3121 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3181 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3241 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3301 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3361 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3421 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3481 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     3541 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3601 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     3661 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     3721 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     3781 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     3841 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     3901 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     3961 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     4021 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     4081 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     4141 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     4201 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     4261 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgtcgt tgctcaggtc
     4321 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     4381 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     4441 cgcacccgtg gccaggaccc aacgctgccc gccgttaacg gccaatacgc aaaccgcctc
     4501 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     4561 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     4621 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     4681 caggaaacag ctatgaccat gattac
//
