LOCUS       sequence-251161-        5378 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of narX under Ptac.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5378)
  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 5378)
  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: pSR348
FEATURES             Location/Qualifiers
     source          1..5378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(97..102,103..118,119..125)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    133..149
                     /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      2007..2078
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2094..2121
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2321..3112
                     /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      3124..3218
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3332..3877
                     /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     3778..3797
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3994..4011
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4050..4127
                     /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             4128..5210
                     /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
                     EDCKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVQALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSV*ARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4147..4166)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      5216..5302
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tgcataattc gtgtagctca aggcgcactc ccgttctgga taatgttttt tgcgccgaca
       61 tcataacggt tctggctaat attctgaaat gagctgttga caattaatca tcggctcgta
      121 taatgtgtgg aattgtgagc ggataacaat ttcacacagc actctcctaa tttaaatcgc
      181 acaactcaca tgcttaaacg ttgtctctct ccgctcaccc tggttaatca ggttgcgctt
      241 attgtgttgc tttctactgc tattggactg gcagggatgg cggtttctgg ctggctggtg
      301 caaggcgttc agggcagcgc ccatgcgatc aacaaagcgg gatcgctgcg catgcaaagt
      361 taccgtctgt tggcggcagt gccattaagc gagaaagaca agcccttaat taaagagatg
      421 gaacaaacgg catttagcgc cgagttgact cgagcagcag aacgagacgg acaactggcg
      481 caattacagg gtttacaaga ttactggcgt aatgaactga tccctgcgct gatgcgtgca
      541 caaaaccgag aaacggtgtc agcggatgtc agccagtttg ttgccgggct tgatcaactg
      601 gtatctggtt ttgaccgcac cacggaaatg cgcatcgaga cagtggtact ggtccatcgg
      661 gtaatggcgg tatttatggc acttttactg gtgttcacta ttatctggtt gcgggcgcga
      721 ctgctacaac cgtggcggca actgctggca atggcgagtg ccgtcagtca tcgcgatttt
      781 acccaacgcg caaacatcag cgggcgcaac gaaatggcga tgcttggaac tgcgttgaac
      841 aatatgtctg cagaactggc cgaaagttat gccgtacttg agcagcgggt tcaggagaaa
      901 accgccgggc tggagcataa aaatcagatc ctctcttttt tatggcaggc taaccgccgt
      961 ttgcattccc gcgccccgct gtgtgaacgc ctgtcacctg tactcaacgg cttacagaat
     1021 ttaaccctgc tacgtgatat cgaattgcgg gtgtatgaca ctgatgatga agagaatcat
     1081 caggagttta cctgccagcc agatatgact tgtgatgata aaggctgcca gctctgcccg
     1141 cgcggcgtat tacccgttgg tgatcgcggc acgaccctga agtggcggct ggctgactct
     1201 catacgcagt acggtatttt gctggcgacc ctgccgcagg ggcgtcatct tagccatgat
     1261 caacaacaac tggtggatac cctggttgaa caactcaccg ccacgctggc gctggatcgc
     1321 catcaggaac gtcagcaaca gttgatcgtg atggaagagc gtgccaccat tgcgcgcgaa
     1381 ctgcatgatt ctattgccca atctctctct tgcatgaaga tgcaggtgag ttgtttacag
     1441 atgcagggcg atgcgctgcc agaaagcagc cgcgaactgt taagtcagat ccgtaacgaa
     1501 ctgaatgcat cctgggcgca gttgcgtgaa ttgctcacca cattccgctt gcagctcacc
     1561 gagcctggat tacgtccggc gctggaggcg agttgcgaag agtacagcgc caaatttggc
     1621 ttcccggtga agctggatta tcaattgccg cctcgcctgg tgccttcgca tcaggcaatc
     1681 cacttgttgc aaattgcccg tgaggcatta agtaacgccc tcaaacattc gcaagcgagt
     1741 gaagtcgtgg tgacggtggc gcaaaacgat aatcaggtca aactgaccgt ccaggataac
     1801 ggctgcggcg tgcctgaaaa tgccatccgc agcaatcact acggcatgat aataatgcgc
     1861 gatcgtgcgc aaagtttacg aggcgattgc cgcgtccgcc gtcgtgaatc aggtggcacc
     1921 gaagtggtgg tcacctttat tcccgaaaaa actttcacag acgtccaagg agatacccat
     1981 gagtaataat aatctagacc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     2041 cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac
     2101 cttcgggtgg gcctttctgc gtttataggt accaggggat cctctagagt cgacctgcag
     2161 gcatgcaagc ttagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg
     2221 gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg
     2281 atcaagatct gatcaagaga caggatgagg atcgtttcgt atgagggaag cggtgatcgc
     2341 cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac
     2401 gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga
     2461 tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat
     2521 caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga
     2581 agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact
     2641 gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat
     2701 cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg
     2761 tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa
     2821 tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt
     2881 gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt
     2941 cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc
     3001 tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga
     3061 agaatttgtc cactacgtga aaggcgagat caccaaggta gtaggcaaat aagagctcgc
     3121 ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa
     3181 cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagcactagt aacaacttat
     3241 atcgtatggg gctgacttca ggtgctacat ttgaagagat aaattgcact gaaatctagt
     3301 aatattttat ctgattaata agatgatctt cttgagatcg ttttggtctg cgcgtaatct
     3361 cttgctctga aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt ctctgagcta
     3421 ccaactcttt gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa cttgtccttt
     3481 cagtttagcc ttaaccggcg catgacttca agactaactc ctctaaatca attaccagtg
     3541 gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg actcaagacg atagttaccg
     3601 gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca tacagtccag cttggagcga
     3661 actgcctacc cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg
     3721 aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc acgagggagc cgccaggggg
     3781 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac cactgatttg agcgtcagat
     3841 ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac ggctttgccg cggccctctc
     3901 acttccctgt taagtatctt cctggcatct tccgggaaat ctccgccccg ttcgtaagcc
     3961 atttccgctc gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa
     4021 tatatcccga agcggcatgc atttacgttg acaccatcga atggtgcaaa acctttcgcg
     4081 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
     4141 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     4201 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     4261 tgaattacat tccaaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     4321 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     4381 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     4441 aagactgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     4501 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttcagg
     4561 cgttatttct tgatgtctct gaccagacac caatcaacag tattattttc tcccatgaag
     4621 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
     4681 tagcgggccc attaagttct gtctaggcgc gtctgcgtct ggctggctgg cataaatatc
     4741 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
     4801 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tccaactgcg atgctggttg
     4861 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
     4921 gtgcggatat ctcggtagtg ggatacgacg atacagaaga cagctcatgt tatatcccgc
     4981 cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     5041 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
     5101 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     5161 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga ggcatcaaat
     5221 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     5281 cgctctcctg agtaggacaa atccgccgcc ctagacctag ggcgttcggc tgcggcgagc
     5341 ggtatcagct cactcaaagg cggtaatacg taaatcac
//
