LOCUS       sequence-238291-        5088 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of uhpB under Ptac.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5088)
  AUTHORS   Schmidl SR, Ekness F, Sofjan K, Daeffler KN, Brink KR, Landry BP, 
            Gerhardt KP, Dyulgyarov N, Sheth RU, Tabor JJ
  TITLE     Rewiring bacterial two-component systems by modular DNA-binding 
            domain swapping.
  JOURNAL   Nat Chem Biol. 2019 Jul;15(7):690-698. doi: 
            10.1038/s41589-019-0286-6. Epub 2019 May 20.
  PUBMED    31110305
REFERENCE   2  (bases 1 to 5088)
  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: 
            "10.1038/s41589-019-0286-6"; journalName: "Nat Chem Biol"; date: 
            "2019-07"; volume: "15"; issue: "7"; pages: "690-698"
COMMENT     Sequence Label: pSR305
FEATURES             Location/Qualifiers
     source          1..5088
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..100
                     /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."
     CDS             101..1183
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(120..139)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      1189..1275
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(1448..1453,1454..1469,1470..1476)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1484..1500
                     /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)."
     terminator      3068..3139
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3155..3182
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             3382..4173
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     terminator      4185..4279
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      4393..4938
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4839..4858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5055..5072
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgaagcggca tgcatttacg ttgacaccat cgaatggtgc aaaacctttc gcggtatggc
       61 atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata
      121 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
      181 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
      241 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
      301 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
      361 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
      421 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
      481 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
      541 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac
      601 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
      661 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
      721 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
      781 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
      841 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
      901 tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac
      961 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
     1021 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
     1081 aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1141 gcagctggca cgacaggttt cccgactgga aagcgggcag tgaggcatca aataaaacga
     1201 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     1261 ctgagtagga caaatccgcc gccctagacc tagggcgttc ggctgcggcg agcggtatca
     1321 gctcactcaa aggcggtaat acgtaaatca ctgcataatt cgtgtcgctc aaggcgcact
     1381 cccgttctgg ataatgtttt ttgcgccgac atcataacgg ttctggctaa tattctgaaa
     1441 tgagctgttg acaattaatc atcggctcgt ataatgtgtg gaattgtgag cggataacaa
     1501 tttcacacat actattacta ttagtttatc ttaccagata aaagatgaag acgttgttct
     1561 cccgcttaat taccgttatt gcctgctttt ttatcttctc tgccgcatgg ttttgcctgt
     1621 ggagtatcag cctgcatctg gttgagcgcc ctgatatggc ggtgctgtta tttccgtttg
     1681 gtctgcgtct ggggctaatg ctgcaatgcc cgcgcggata ctggcccgta ttgctgggcg
     1741 cggagtggct gctgatttac tggctaacgc aggcggtcgg tttaacccat tttccgttat
     1801 tgatgatcgg tagtttactg acgttactgc ccgtagcgct gatctcgcgc tatcgccatc
     1861 agcgtgactg gcgcaccttg ctgttacagg gggcggcgtt aacggcggcg gcgttgttgc
     1921 agtcgctgcc ctggctttgg cacggcaaag agtcgtggaa tgcgctgttg ctgactttaa
     1981 ctggcggcct gacgctggcc ccgatatgtc tggtgttctg gcactatctc gccaataaca
     2041 cctggctgcc gctcggtccg tcactggttt ctcagccaat caactggcgc gggcgacatc
     2101 tggtctggta cttgctgctg tttgttatca gtctctggct ccagttggga ttgccggacg
     2161 aactgtcgcg ctttacgcca ttctgtctgg cgctgccgat tatcgcgctg gcctggcact
     2221 atggttggca aggggcgctg attgcgacgt tgatgaacgc catcgcgctg atcgccagtc
     2281 aaacctggcg cgatcatccg gtggatttat tgctctcgct gctggtgcaa agtctgacag
     2341 ggttgttgct tggcgctggc atccagcggt tgcgtgaact taaccagtcg ctgcaaaagg
     2401 aactggcgcg caatcagcat ctggctgaac ggttgctgga aaccgaagag agcgtgcgcc
     2461 gtgatgtggc gcgtgagctg catgatgata tcggtcagac catcactgct attcgtactc
     2521 aggcgggcat tgttcagcgg ctggcggcag ataacgccag cgtgaagcag agcgggcagc
     2581 tcatcgaaca actatcgctg ggcgtttacg acgcggtgcg ccgtttgttg ggtcggttac
     2641 gtccgcgcca gttggatgat ctcaccctgg agcaggccat ccgctcactg atgcgggaaa
     2701 tggagctgga agggcgcggt attgtcagcc atctcgaatg gcgaatcgat gaatcagcgt
     2761 taagcgaaaa ccagcgcgtg acgctgtttc gtgtctgcca ggaagggctg aacaacattg
     2821 tgaaacatgc tgatgccagc gcggtcaccc tgcaaggctg gcagcaggat gaacggttga
     2881 tgctggttat tgaagacgat ggcagcggtt tgccgccggg ttccgggcaa caaggttttg
     2941 gcctcaccgg aatgcgcgag cgcgtaacgg cgctgggtgg cacattacac atttcctgtc
     3001 tgcacggcac gcgtgtcagc gtttctctac ctcaacgcta tgtctaataa taatctagac
     3061 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     3121 tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg
     3181 cgtttatagg tacccgggga tcctctagag tcgacctgca ggcatgcaag cttagcaagc
     3241 gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac
     3301 tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagatc tgatcaagag
     3361 acaggatgag gatcgtttcg catgagggaa gcggtgatcg ccgaagtatc gactcaacta
     3421 tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg
     3481 tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg
     3541 gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact
     3601 tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac
     3661 gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag
     3721 cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc
     3781 ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc
     3841 tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg
     3901 aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt
     3961 tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg
     4021 gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga
     4081 caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg
     4141 aaaggcgaga tcaccaaggt agtcggcaaa taagagctcg cttggactcc tgttgataga
     4201 tccagtaatg acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc
     4261 gttttttatt ggtgagaatc caagcactag taacaactta tatcgtatgg ggctgacttc
     4321 aggtgctaca tttgaagaga taaattgcac tgaaatctag taatatttta tctgattaat
     4381 aagatgatct tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa
     4441 aaccgccttg cagggcggtt tttcgaaggt tctctgagct accaactctt tgaaccgagg
     4501 taactggctt ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc
     4561 gcatgacttc aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt
     4621 ttgcatgtct ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4681 actgaacggg gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga
     4741 gtgtcaggcg tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga
     4801 aaggcaggaa caggagagcg cacgagggag ccgccagggg gaaacgcctg gtatctttat
     4861 agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg
     4921 gggcggagcc tatggaaaaa cggctttgcc gcggccctct cacttccctg ttaagtatct
     4981 tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc catttccgct cgccgcagtc
     5041 gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga atatatcc
//
