LOCUS       sequence-104837-        6282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CcaS, Ho1, and PcyA constitutively.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6282)
  AUTHORS   Schmidl SR, Sheth RU, Wu A, Tabor JJ
  TITLE     Refactoring and optimization of light-switchable escherichia coli 
            two-component systems.
  JOURNAL   ACS Synth Biol. 2014 Nov 21;3(11):820-31. doi: 10.1021/sb500273n. 
            Epub 2014 Oct 11.
  PUBMED    25250630
REFERENCE   2  (bases 1 to 6282)
  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: "10.1021/sb500273n";
            journalName: "ACS Synth Biol"; date: "2014-11-21- 21"; volume: "3"; 
            issue: "11"; pages: "820-31"
COMMENT     Sequence Label: pSR43.6r
FEATURES             Location/Qualifiers
     source          1..6282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..35
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     terminator      2353..2424
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2440..2467
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2584..2618
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     RBS             2635..2646
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     RBS             3378..3389
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      4157..4228
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4244..4271
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             4471..5262
                     /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      5274..5368
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      5482..6027
                     /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     5928..5947
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6144..6161
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tttacggcta gctcagtcct aggtatagtg ctagctatag aggttaaaat cttaatagga
       61 ggaaggcgca atgggcaaat ttctaattcc aatcgaattt gtttttctgg cgatcgccat
      121 gacctgttat ttatggcaca gacaaaacca agaacgccgc aggattgaaa ttagcatcaa
      181 gcaacaaacc caacgggaac gatttattaa ccaaattacc caacatatcc gccaatcttt
      241 aaacttggaa acggttttaa ataccaccgt cgctgaagtt aaaaccctgt tgcaagttga
      301 tcgagttcta atttatcgca tttggcaaga tggcacgggc agcgccatta cggaatcggt
      361 gaatgccaat tatcctagta ttttagggcg gaccttttcc gatgaagttt ttcccgttga
      421 ataccatcaa gcctacacca aaggtaaagt acgggccatt aatgacattg accaggatga
      481 catagagatt tgcctagctg atttcgtcaa acaatttggc gtgaaatcaa aattagtagt
      541 gcccattctt caacataatc gtgcttcttc cctagataat gaatcagaat ttccctatct
      601 ttgggggctg ttaattaccc atcaatgtgc ttttacccgg ccatggcaac cgtgggaagt
      661 ggagttaatg aaacagctag ccaatcaggt cgcgatcgcc atccaacaat cggaattata
      721 tgagcaatta cagcaactca ataaagattt ggaaaaccga gtcgaaaaac gcacccagca
      781 acttgccgcc accaatcaat ccctaagaat ggaaatcagt gagcgacaaa aaacggaagc
      841 cgctctccgc cacactaacc atactctgca atccctgatt gcggcctccc ccaggggtat
      901 ttttaccctt aatttagcag accaaattca gatttggaat cctacagcag aacgtatttt
      961 tggttggaca gaaacagaaa ttattgccca tccagaatta ttaacatcca acattttgct
     1021 ggaagattat cagcaattta aacagaaagt tttatcaggc atggtttccc ctagcctaga
     1081 attaaaatgt caaaaaaaag atggtagttg gattgaaatt gtcctttccg ctgctcccct
     1141 attggatagt gaagaaaata ttgccggatt ggtggcggtt gtcgccgata ttaccgagca
     1201 aaagcggcag gcagaacaaa ttcgtttgct acaatccgtt gtggttaata ctaatgatgc
     1261 ggtggtgatt acggaagcgg agcccattga tgatcccggg ccgagaattc tctatgtcaa
     1321 tgaagcattt actaaaatca ccggttatac tgctgaagaa atgctaggca aaaccccccg
     1381 agttttacag ggaccaaaaa ctagtcgcac tgaattagat agggtgcggc aagccattag
     1441 tcaatggcaa tcagttaccg ttgaagtgat taattatcgt aaggatggca gtgagttttg
     1501 ggtggaattt agtctggtgc ccgttgccaa taaaacaggt ttttacaccc attggattgc
     1561 tgtgcaaagg gatgtcactg agcgccgacg cacggaggaa gtccgcctag ctttagaacg
     1621 ggaaaaagaa ttaagccgcc taaaaactcg ttttttctcc atggcttccc atgaatttcg
     1681 tactcccctc agtacggcct tagctgctgc ccaattactg gaaaattctg aagtggcctg
     1741 gcttgatccc gataagcgta gccggaactt acaccgtatt caaaattccg tgaaaaatat
     1801 ggtacagctc ctggatgata ttttaatcat taaccgtgcc gaagcgggca aattggaatt
     1861 taatcctaat tggttagatt tgaaattatt gttccagcaa tttatcgaag aaattcaatt
     1921 aagtgtcagt gaccaatatt attttgactt tatttgtagc gctcaagata cgaaggcatt
     1981 ggtggatgaa aggttagtgc ggtctatttt atctaatctg ttatctaatg cgattaaata
     2041 ctctcccggg ggagggcaga ttaaaattgc cctaagccta gattcggaac agattatttt
     2101 tgaagtcacc gaccagggca ttggcatttc gccagaggac caaaagcaaa tttttgaacc
     2161 ctttcatcgg ggcaaaaatg tcagaaatat tacgggaaca ggactcggtt taatggttgc
     2221 caagaaatgt gttgacttac acagtggcag tatcttgcta aaaagtgcag ttgaccaggg
     2281 aacaacagtt actatctgtt taaaacgcta taaccatttg cctcgagctt agtaataatc
     2341 tagaccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     2401 tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct
     2461 ttctgcgttt atagtaaatc actgcataat tcgtgtcgat taaaccgcct aagtccccca
     2521 ggaaaggggg atataacagt atagattttg tcagccttca gcttggcttt accgtcaaaa
     2581 aaactgacag ctagctcagt cctaggtata atgctagctc acacagaatt cattaaagag
     2641 gagaaaggta ccatgagtgt caacttagct tcccagttgc gggaagggac gaaaaaatcc
     2701 cactccatgg cggagaacgt cggctttgtc aaatgcttcc tcaagggcgt tgtcgagaaa
     2761 aattcctacc gtaagctggt tggcaatctc tactttgtct acagtgccat ggaagaggaa
     2821 atggcaaaat ttaaggacca tcccatcctc agccacattt acttccccga actcaaccgc
     2881 aaacaaagcc tagagcaaga cctgcaattc tattacggct ccaactggcg gcaagaagtg
     2941 aaaatttctg ccgctggcca agcctatgtg gaccgagtcc ggcaagtggc cgctacggcc
     3001 cctgaattgt tggtggccca ttcctacacc cgttacctgg gggatctttc cggcggtcaa
     3061 attctcaaga aaattgccca aaatgccatg aatctccacg atggtggcac agctttctat
     3121 gaatttgccg acattgatga cgaaaaggct tttaaaaata cctaccgtca agctatgaat
     3181 gatctgccca ttgaccaagc caccgccgaa cggattgtgg atgaagccaa tgacgccttt
     3241 gccatgaaca tgaaaatgtt caacgaactt gaaggcaacc tgatcaaggc gatcggcatt
     3301 atggtgttca acagcctcac ccgtcgccgc agtcaaggca gcaccgaagt tggcctcgcc
     3361 acctccgaag gctagttaaa gaggagaaag gatccatggc cgtcactgat ttaagtttga
     3421 ccaattcttc cctgatgccc acgttgaacc cgatgattca acagttggcc ctggcgatcg
     3481 ccgctagttg gcaaagttta cccctcaagc cctatcaatt gccggaggat ttgggctacg
     3541 tagaaggccg cctggaaggg gaaaagttag tgattgaaaa tcggtgctac caaacgcccc
     3601 agtttcgcaa aatgcatttg gagttggcca aggtgggcaa agggttggat attctccact
     3661 gtgtaatgtt tcctgagcct ttatacggtc tacctttgtt tggctgtgac attgtggccg
     3721 gccccggtgg agtaagtgcg gctattgcgg atctatcccc cacccaaagc gatcgccaat
     3781 tgcccgcagc gtaccaaaaa tcattggcag agctaggcca gccagaattt gagcaacaac
     3841 gggaattgcc cccctgggga gaaatatttt ctgaatattg tttattcatc cgtcccagca
     3901 atgtcactga agaagaaaga tttgtacaaa gggtagtgga ctttttgcaa attcattgtc
     3961 accaatccat cgttgccgaa cccttgtctg aagctcaaac tttggagcac cgtcaggggc
     4021 aaattcatta ctgccaacaa caacagaaaa atgataaaac ccgtcgggta ctggaaaaag
     4081 cttttgggga agcttgggcg gaacggtata tgagccaagt cttatttgat gttatccaat
     4141 aatctagacc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     4201 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
     4261 gcctttctgc gtttataggt acccggggat cctctagagt cgacctgcag gcatgcaagc
     4321 ttagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc
     4381 aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagatct
     4441 gatcaagaga caggatgagg atcgtttcgc atgagggaag cggtgatcgc cgaagtatcg
     4501 actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc
     4561 gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg
     4621 ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt
     4681 ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt
     4741 gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga
     4801 gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat
     4861 ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg
     4921 gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta
     4981 acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg
     5041 tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac
     5101 tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct
     5161 tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc
     5221 cactacgtga aaggcgagat caccaaggta gtcggcaaat aagagctcgc ttggactcct
     5281 gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg
     5341 ccgccgggcg ttttttattg gtgagaatcc aagcactagt aacaacttat atcgtatggg
     5401 gctgacttca ggtgctacat ttgaagagat aaattgcact gaaatctaga aatattttat
     5461 ctgattaata agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga
     5521 aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt ctctgagcta ccaactcttt
     5581 gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc
     5641 ttaaccggcg catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca
     5701 gtggtgcttt tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc
     5761 agcggtcgga ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc
     5821 cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg
     5881 gtaaaccgaa aggcaggaac aggagagcgc acgagggagc cgccaggggg aaacgcctgg
     5941 tatctttata gtcctgtcgg gtttcgccac cactgatttg agcgtcagat ttcgtgatgc
     6001 ttgtcagggg ggcggagcct atggaaaaac ggctttgccg cggccctctc acttccctgt
     6061 taagtatctt cctggcatct tccaggaaat ctccgccccg ttcgtaagcc atttccgctc
     6121 gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa tatatcccct
     6181 aggtggagtc tggcctcaaa tacaaagtct gcaaagtctc gaaatattaa accagagaat
     6241 aaataattaa acgccaaaat atttctagca aaaaccccag tt
//
