LOCUS       sequence-238305-        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        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      3358..3429
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3445..3472
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             3672..4463
                     /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      4475..4569
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      4683..5228
                     /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     5129..5148
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5345..5362
                     /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 tttcacacag cactctccta atttaaatcg cacaactcac atgcttaaac gttgtctctc
     1561 tccgctcacc ctggttaatc aggttgcgct tattgtgttg ctttctactg ctattggact
     1621 ggcagggatg gcggtttctg gctggctggt gcaaggcgtt cagggcagcg cccatgcgat
     1681 caacaaagcg ggatcgctgc gcatgcaaag ttaccgtctg ttggcggcag tgccattaag
     1741 cgagaaagac aagcccttaa ttaaagagat ggaacaaacg gcatttagcg ccgagttgac
     1801 tcgagcagca gaacgagacg gacaactggc gcaattacag ggtttacaag attactggcg
     1861 taatgaactg atccctgcgc tgatgcgtgc acaaaaccga gaaacggtgt cagcggatgt
     1921 cagccagttt gttgccgggc ttgatcaact ggtatctggt tttgaccgca ccacggaaat
     1981 gcgcatcgag acagtggtac tggtccatcg ggtaatggcg gtatttatgg cacttttact
     2041 ggtgttcact attatctggt tgcgggcgcg actgctacaa ccgtggcggc aactgctggc
     2101 aatggcgagt gccgtcagtc atcgcgattt tacccaacgc gcaaacatca gcgggcgcaa
     2161 cgaaatggcg atgcttggaa ctgcgttgaa caatatgtct gcagaactgg ccgaaagtta
     2221 tgccgtactt gagcagcggg ttcaggagaa aaccgccggg ctggagcata aaaatcagat
     2281 cctctctttt ttatggcagg ctaaccgccg tttgcattcc cgcgccccgc tgtgtgaacg
     2341 cctgtcacct gtactcaacg gcttacagaa tttaaccctg ctacgtgata tcgaattgcg
     2401 ggtgtatgac actgatgatg aagagaatca tcaggagttt acctgccagc cagatatgac
     2461 ttgtgatgat aaaggctgcc agctctgccc gcgcggcgta ttacccgttg gtgatcgcgg
     2521 cacgaccctg aagtggcggc tggctgactc tcatacgcag tacggtattt tgctggcgac
     2581 cctgccgcag gggcgtcatc ttagccatga tcaacaacaa ctggtggata ccctggttga
     2641 acaactcacc gccacgctgg cgctggatcg ccatcaggaa cgtcagcaac agttgatcgt
     2701 gatggaagag cgtgccacca ttgcgcgcga actgcatgat tctattgccc aatctctctc
     2761 ttgcatgaag atgcaggtga gttgtttaca gatgcagggc gatgcgctgc cagaaagcag
     2821 ccgcgaactg ttaagtcaga tccgtaacga actgaatgca tcctgggcgc agttgcgtga
     2881 attgctcacc acattccgct tgcagctcac cgagcctgga ttacgtccgg cgctggaggc
     2941 gagttgcgaa gagtacagcg ccaaatttgg cttcccggtg aagctggatt atcaattgcc
     3001 gcctcgcctg gtgccttcgc atcaggcaat ccacttgttg caaattgccc gtgaggcatt
     3061 aagtaacgcc ctcaaacatt cgcaagcgag tgaagtcgtg gtgacggtgg cgcaaaacga
     3121 taatcaggtc aaactgaccg tccaggataa cggctgcggc gtgcctgaaa atgccatccg
     3181 cagcaatcac tacggcatga taataatgcg cgatcgtgcg caaagtttac gaggcgattg
     3241 ccgcgtccgc cgtcgtgaat caggtggcac cgaagtggtg gtcaccttta ttcccgaaaa
     3301 aactttcaca gacgtccaag gagataccca tgagtaataa taatctagac caggcatcaa
     3361 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     3421 aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatagg
     3481 tacccgggga tcctctagag tcgacctgca ggcatgcaag cttagcaagc gaaccggaat
     3541 tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac tggatggctt
     3601 tcttgccgcc aaggatctga tggcgcaggg gatcaagatc tgatcaagag acaggatgag
     3661 gatcgtttcg catgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag
     3721 ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg
     3781 cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa
     3841 ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc
     3901 ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca
     3961 ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca
     4021 ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa
     4081 aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg
     4141 ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc
     4201 ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg
     4261 cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc
     4321 cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag
     4381 atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga
     4441 tcaccaaggt agtcggcaaa taagagctcg cttggactcc tgttgataga tccagtaatg
     4501 acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt
     4561 ggtgagaatc caagcactag taacaactta tatcgtatgg ggctgacttc aggtgctaca
     4621 tttgaagaga taaattgcac tgaaatctag taatatttta tctgattaat aagatgatct
     4681 tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg
     4741 cagggcggtt tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt
     4801 ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc
     4861 aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct
     4921 ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg
     4981 gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg
     5041 tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa
     5101 caggagagcg cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg
     5161 ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc
     5221 tatggaaaaa cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc
     5281 ttccaggaaa tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga
     5341 gcgtagcgag tcagtgagcg aggaagcgga atatatcc
//
