LOCUS       sequence-259021-        4755 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector with aTc inducible M. laminosus Fd1 mutant with the estrogen 
            recepor-ligand binding domain fused between amino acids 25 and 26 
            with a C-terminal fused mRFP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4755)
  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 4755)
  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: pBW007
FEATURES             Location/Qualifiers
     source          1..4755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     59..77
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(98..116,117..122,123..139,140..145,146..171)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    98..116
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    123..141
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1373..2047
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     terminator      2077..2148
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2164..2191
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2243..2277
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2312..2938
                     /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(2388..2407)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2944..3030
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3195..3783)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3275..3294)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3871..3965)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3986..4645)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4560..4579
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 attttcgcca gatatcgacg tctaagaaac cattattatc atgacattaa cctataaaaa
       61 taggcgtatc acgaggccct ttcgtcttga cctcgagtcc ctatcagtga tagagattga
      121 catccctatc agtgatagag atactgagca catcagcagg acgcactgac ctaaaaatcc
      181 gatccctctt tagttctcaa tcatggcaac ctataaagtt acacttatta acgaagctga
      241 aggtctaaac aaaacaattg aagttccaga tgatcagtac attgctgcag ctgggggcgg
      301 aggttctgga ggtggtgggt ccaagaagaa ttcactggcg ttgtcgctta cggctgacca
      361 aatggtttct gctctgcttg acgccgaacc tccgatcctg tactctgaat acgacccgac
      421 tcgtcctttt tcagaggcga gtatgatggg gctgcttacg aatctggcag accgcgagtt
      481 ggtacatatg attaattggg ctaaacgtgt gccgggtttc gtggacttga ccctgcatga
      541 ccaggtccac ctgttagaat gcgcctggtt agagatttta atgatcggac ttgtctggcg
      601 tagcatggaa caccccggaa aactgttatt tgcccctaac ttgctgctgg accgcaacca
      661 ggggaagtgt gtcgaaggaa tggtggaaat tttcgacatg cttcttgcca cttcctcgcg
      721 ttttcgtatg atgaatttac agggagaaga gttcgtttgt ttaaagtcaa tcatcttact
      781 gaatagcggg gtttacacat tcttatcttc cactcttaaa tcactggagg agaaagacca
      841 tattcaccgt gtcctggaca agatcacaga tactcttatc catcttatgg cgaaggcagg
      901 cttaacactt caacagcaac atcagcgcct tgctcagctt ctgttgattt taagccatat
      961 ccgtcacatg tcaaacaagg gtatggaaca cctttacagt atgaagtgta agaacgtcgt
     1021 tccattgtat gatttgttac tggaaatgct tgacgctcac cgtctgcatg cacctggtgg
     1081 cggtggctct ggtggaggcg ggagcgcagc tgcatacatt ttagatgcgg ctgaagaagc
     1141 tggtatagac ttaccgtatt cttgtcgtgc tggtgcttgc tctacctgtg ctggtaaact
     1201 catatccggt actgtcgatc aaagcgatca gtcattttta gacgacgatc aaattgaagc
     1261 tggatacgtt ctcacctgcg ttgcttaccc aacttccgat tgcgtcatcg aaacacacaa
     1321 agaagaagag ctttacgggg gctctggagg ttctggaggt tctggtgggt ccgcgagtag
     1381 cgaagacgtt atcaaagagt tcatgcgttt caaagttcgt atggaaggtt ccgttaacgg
     1441 tcacgagttc gaaatcgaag gtgaaggtga aggtcgtccg tacgaaggta cccagaccgc
     1501 taaactgaaa gttaccaaag gtggtccgct gccgttcgct tgggacatcc tgtccccgca
     1561 gttccagtac ggttccaaag cttacgttaa acacccggct gacatcccgg actacctgaa
     1621 actgtccttc ccggaaggtt tcaaatggga acgtgttatg aacttcgaag acggtggtgt
     1681 tgttaccgtt acccaggact cctccctgca agacggtgag ttcatctaca aagttaaact
     1741 gcgtggtacc aacttcccgt ccgacggtcc ggttatgcag aaaaaaacca tgggttggga
     1801 agcttccacc gaacgtatgt acccggaaga cggtgctctg aaaggtgaaa tcaaaatgcg
     1861 tctgaaactg aaagacggtg gtcactacga cgctgaagtt aaaaccacct acatggctaa
     1921 aaaaccggtt cagctgccgg gtgcttacaa aaccgacatc aaactggaca tcacctccca
     1981 caacgaagac tacaccatcg ttgaacagta cgaacgtgct gaaggtcgtc actccaccgg
     2041 tgcttgagaa ttctgataat aatctagacc aggcatcaaa taaaacgaaa ggctcagtcg
     2101 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac
     2161 actggctcac cttcgggtgg gcctttctgc gtttataggt acccgtggat cctctagagt
     2221 cgacctgcag gcatgcaagc tttttacggc tagctcagtc ctaggtatag tgctagccca
     2281 gccagagaaa cactctttaa cagggggtag tatgatgtct cgtttagata aaagtaaagt
     2341 gattaacagc gcattagagc tgcttaatga ggtcggaatc gaaggtttaa caacccgtaa
     2401 actcgcccag aagctaggtg tagagcagcc tacattgtat tggcatgtaa aaaataagcg
     2461 ggctttgctc gacgccttag ccattgagat gttagatagg caccatactc acttttgccc
     2521 tttagaaggg gaaagctggc aagatttttt acgtaataac gctaaaagtt ttagatgtgc
     2581 tttactaagt catcgcgatg gagcaaaagt acatttaggt acacggccta cagaaaaaca
     2641 gtatgaaact ctcgaaaatc aattagcctt tttatgccaa caaggttttt cactagagaa
     2701 tgcattatat gcactcagcg cagtggggca ttttacttta ggttgcgtat tggaagatca
     2761 agagcatcaa gtcgctaaag aagaaaggga aacacctact actgatagta tgccgccatt
     2821 attacgacaa gctatcgaat tatttgatca ccaaggtgca gagccagcct tcttattcgg
     2881 ccttgaattg atcatatgcg gattagaaaa acaacttaaa tgtgaaagtg ggtcttaagg
     2941 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     3001 tcggtgaacg ctctcctgag taggacaaat ccgccgccct agacctaggg cgttcggctg
     3061 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3121 aacgcatgca agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3181 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3241 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3301 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3361 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3421 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3481 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3541 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3601 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     3661 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3721 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3781 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3841 taagggattt tggtcatgac tagtgcttgg attctcacca ataaaaaacg cccggcggca
     3901 accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag
     3961 gagtccaagc gagctcgata tcaaattacg ccccgccctg ccactcatcg cagtactgtt
     4021 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
     4081 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
     4141 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     4201 gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     4261 caccataaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
     4321 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
     4381 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacgaaat tccggatgag
     4441 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
     4501 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag
     4561 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
     4621 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact
     4681 caaaaaatac gaccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgg
     4741 gccggccaac gtctc
//
