LOCUS       sequence-258989-        3918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector with aTc inducible M. laminosus Fd1 with a C-terminal fused 
            mRFP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3918)
  AUTHORS   Wu B, Atkinson JT, Kahanda D, Bennett GN, Silberg JJ
  TITLE     Combinatorial design of chemical-dependent protein switches for 
            controlling intracellular electron transfer
  JOURNAL   bioRxiv May 23, 2019
REFERENCE   2  (bases 1 to 3918)
  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"; journalName: "bioRxiv May
            23, 2019"
COMMENT     Sequence Label: pBW005
FEATURES             Location/Qualifiers
     source          1..3918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     8..26
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(47..65,66..71,72..88,89..94,95..120)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    47..65
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    72..90
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             485..1159
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     terminator      1189..1260
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1276..1303
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1355..1389
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             1424..2050
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(1500..1519)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2056..2142
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(2307..2895)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2387..2406)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2983..3077)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3098..3757)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3672..3691
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ctataaaaat aggcgtatca cgaggccctt tcgtcttcac ctcgagtccc tatcagtgat
       61 agagattgac atccctatca gtgatagaga tactgagcac atcagcagga cgcactgacc
      121 taaaaatccg atccctcttt agttctcaat catggcaacc tataaagtta cacttattaa
      181 cgaagctgaa ggtctaaaca aaacaattga agttccagat gatcagtaca ttttagatgc
      241 ggctgaagaa gctggtatag acttaccgta ttcttgtcgt gctggtgctt gctctacctg
      301 tgctggtaaa ctcatatccg gtactgtcga tcaaagcgat cagtcatttt tagacgacga
      361 tcaaattgaa gctggatacg ttctcacctg cgttgcttac ccaacttccg attgcgtcat
      421 cgaaacacac aaagaagaag agctttacgg gggctctgga ggttctggag gttctggtgg
      481 gtccgcgagt agcgaagacg ttatcaaaga gttcatgcgt ttcaaagttc gtatggaagg
      541 ttccgttaac ggtcacgagt tcgaaatcga aggtgaaggt gaaggtcgtc cgtacgaagg
      601 tacccagacc gctaaactga aagttaccaa aggtggtccg ctgccgttcg cttgggacat
      661 cctgtccccg cagttccagt acggttccaa agcttacgtt aaacacccgg ctgacatccc
      721 ggactacctg aaactgtcct tcccggaagg tttcaaatgg gaacgtgtta tgaacttcga
      781 agacggtggt gttgttaccg ttacccagga ctcctccctg caagacggtg agttcatcta
      841 caaagttaaa ctgcgtggta ccaacttccc gtccgacggt ccggttatgc agaaaaaaac
      901 catgggttgg gaagcttcca ccgaacgtat gtacccggaa gacggtgctc tgaaaggtga
      961 aatcaaaatg cgtctgaaac tgaaagacgg tggtcactac gacgctgaag ttaaaaccac
     1021 ctacatggct aaaaaaccgg ttcagctgcc gggtgcttac aaaaccgaca tcaaactgga
     1081 catcacctcc cacaacgaag actacaccat cgttgaacag tacgaacgtg ctgaaggtcg
     1141 tcactccacc ggtgcttgag aattctgata ataatctaga ccaggcatca aataaaacga
     1201 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     1261 tactagagtc acactggctc accttcgggt gggcctttct gcgtttatag gtacccgtgg
     1321 atcctctaga gtcgacctgc aggcatgcaa gctttttacg gctagctcag tcctaggtat
     1381 agtgctagcc cagccagaga aacactcttt aacagggggt agtatgatgt ctcgtttaga
     1441 taaaagtaaa gtgattaaca gcgcattaga gctgcttaat gaggtcggaa tcgaaggttt
     1501 aacaacccgt aaactcgccc agaagctagg tgtagagcag cctacattgt attggcatgt
     1561 aaaaaataag cgggctttgc tcgacgcctt agccattgag atgttagata ggcaccatac
     1621 tcacttttgc cctttagaag gggaaagctg gcaagatttt ttacgtaata acgctaaaag
     1681 ttttagatgt gctttactaa gtcatcgcga tggagcaaaa gtacatttag gtacacggcc
     1741 tacagaaaaa cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc aacaaggttt
     1801 ttcactagag aatgcattat atgcactcag cgcagtgggg cattttactt taggttgcgt
     1861 attggaagat caagagcatc aagtcgctaa agaagaaagg gaaacaccta ctactgatag
     1921 tatgccgcca ttattacgac aagctatcga attatttgat caccaaggtg cagagccagc
     1981 cttcttattc ggccttgaat tgatcatatg cggattagaa aaacaactta aatgtgaaag
     2041 tgggtcttaa ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     2101 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccgcc ctagacctag
     2161 ggcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2221 gaatcagggg ataacgcatg caagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2281 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2341 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2401 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2461 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2521 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2581 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2641 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2701 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     2761 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2821 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2881 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2941 gaaaactcac gttaagggat tttggtcatg actagtgctt ggattctcac caataaaaaa
     3001 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg
     3061 atctatcaac aggagtccaa gcgagctcga tatcaaatta cgccccgccc tgccactcat
     3121 cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
     3181 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
     3241 tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga
     3301 aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat
     3361 aggccaggtt ttcaccataa cacgccacat cttgcgaata tatgtgtaga aactgccgga
     3421 aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg
     3481 tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa
     3541 attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt
     3601 gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat
     3661 aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata
     3721 tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa
     3781 atctcgataa ctcaaaaaat acgaccggta gtgatcttat ttcattatgg tgaaagttgg
     3841 aacctcttac gggccggcca acgtctcatt ttcgccagat atcgacgtct aagaaaccat
     3901 tattatcatg acattaac
//
