LOCUS       sequence-247132-        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"
     terminator      1407..1478
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1494..1521
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             1721..2512
                     /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      2524..2618
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2732..3277
                     /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     3178..3197
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3394..3411
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3450..3527
                     /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             3528..4610
                     /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(3547..3566)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      4616..4702
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(4875..4880,4881..4896,4897..4903)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4911..4927
                     /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)."
ORIGIN
        1 gtgctgttat ttccgtttgg tctgcgtctg gggctaatgc tgcaatgccc gcgcggatac
       61 tggcccgtat tgctgggcgc ggagtggctg ctgatttact ggctaacgca ggcggtcggt
      121 ttaacccatt ttccgttatt gatgatcggt agtttactga cgttactgcc cgtagcgctg
      181 atctcgcgct atcgccatca gcgtgactgg cgcaccttgc tgttacaggg ggcggcgtta
      241 acggcggcgg cgttgttgca gtcgctgccc tggctttggc acggcaaaga gtcgtggaat
      301 gcgctgttgc tgactttaac tggcggcctg acgctggccc cgatatgtct ggtgttctgg
      361 cactatctcg ccaataacac ctggctgccg ctcggtccgt cactggtttc tcagccaatc
      421 aactggcgcg ggcgacatct ggtctggtac ttgctgctgt ttgttatcag tctctggctc
      481 cagttgggat tgccggacga actgtcgcgc tttacgccat tctgtctggc gctgccgatt
      541 atcgcgctgg cctggcacta tggttggcaa ggggcgctga ttgcgacgtt gatgaacgcc
      601 atcgcgctga tcgccagtca aacctggcgc gatcatccgg tggatttatt gctctcgctg
      661 ctggtgcaaa gtctgacagg gttgttgctt ggcgctggca tccagcggtt gcgtgaactt
      721 aaccagtcgc tgcaaaagga actggcgcgc aatcagcatc tggctgaacg gttgctggaa
      781 accgaagaga gcgtgcgccg tgatgtggcg cgtgagctgc atgatgatat cggtcagacc
      841 atcactgcta ttcgtactca ggcgggcatt gttcagcggc tggcggcaga taacgccagc
      901 gtgaagcaga gcgggcagct catcgaacaa ctatcgctgg gcgtttacga cgcggtgcgc
      961 cgtttgttgg gtcggttacg tccgcgccag ttggatgatc tcaccctgga gcaggccatc
     1021 cgctcactga tgcgggaaat ggagctggaa gggcgcggta ttgtcagcca tctcgaatgg
     1081 cgaatcgatg aatcagcgtt aagcgaaaac cagcgcgtga cgctgtttcg tgtctgccag
     1141 gaagggctga acaacattgt gaaacatgct gatgccagcg cggtcaccct gcaaggctgg
     1201 cagcaggatg aacggttgat gctggttatt gaagacgatg gcagcggttt gccgccgggt
     1261 tccgggcaac aaggttttgg cctcaccgga atgcgcgagc gcgtaacggc gctgggtggc
     1321 acattacaca tttcctgtct gcacggcacg cgtgtcagcg tttctctacc tcaacgctat
     1381 gtctaataat aatctagacc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     1441 cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac
     1501 cttcgggtgg gcctttctgc gtttataggt acccggggat cctctagagt cgacctgcag
     1561 gcatgcaagc ttagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg
     1621 gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg
     1681 atcaagatct gatcaagaga caggatgagg atcgtttcgt atgagggaag cggtgatcgc
     1741 cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac
     1801 gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga
     1861 tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat
     1921 caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga
     1981 agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact
     2041 gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat
     2101 cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg
     2161 tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa
     2221 tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt
     2281 gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt
     2341 cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc
     2401 tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga
     2461 agaatttgtc cactacgtga aaggcgagat caccaaggta gtcggcaaat aagagctcgc
     2521 ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa
     2581 cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagcactagt aacaacttat
     2641 atcgtatggg gctgacttca ggtgctacat ttgaagagat aaattgcact gaaatctagt
     2701 aatattttat ctgattaata agatgatctt cttgagatcg ttttggtctg cgcgtaatct
     2761 cttgctctga aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt ctctgagcta
     2821 ccaactcttt gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa cttgtccttt
     2881 cagtttagcc ttaaccggcg catgacttca agactaactc ctctaaatca attaccagtg
     2941 gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg actcaagacg atagttaccg
     3001 gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca tacagtccag cttggagcga
     3061 actgcctacc cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg
     3121 aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc acgagggagc cgccaggggg
     3181 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac cactgatttg agcgtcagat
     3241 ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac ggctttgccg cggccctctc
     3301 acttccctgt taagtatctt cctggcatct tccgggaaat ctccgccccg ttcgtaagcc
     3361 atttccgctc gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa
     3421 tatatcccga agcggcatgc atttacgttg acaccatcga atggtgcaaa acctttcgcg
     3481 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
     3541 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     3601 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     3661 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     3721 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     3781 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     3841 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     3901 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
     3961 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
     4021 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
     4081 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc
     4141 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
     4201 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg
     4261 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
     4321 gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc
     4381 cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     4441 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
     4501 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     4561 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga ggcatcaaat
     4621 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     4681 cgctctcctg agtaggacaa atccgccgcc ctagacctag ggcgttcggc tgcggcgagc
     4741 ggtatcagct cactcaaagg cggtaatacg taaatcactg cataattcgt gtcgctcaag
     4801 gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc tggctaatat
     4861 tctgaaatga gctgttgaca attaatcatc ggctcgtata atgtgtggaa ttgtgagcgg
     4921 ataacaattt cacacatact attactatta gtttatctta ccagataaaa gatgaagacg
     4981 ttgttctccc gcttaattac cgttattgcc tgctttttta tcttctctgc cgcatggttt
     5041 tgcctgtgga gtatcagcct gcatctggtt gagcgccctg atatggcg
//
