LOCUS       sequence-259074-        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 55 and 56 
            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: pBW015
FEATURES             Location/Qualifiers
     source          1..4755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             682..1356
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     terminator      1386..1457
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1473..1500
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1552..1586
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             1621..2247
                     /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(1697..1716)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2253..2339
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(2504..3092)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2584..2603)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3180..3274)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3295..3954)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3869..3888
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     4123..4141
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(4162..4180,4181..4186,4187..4203,4204..4209,
                     4210..4235)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    4162..4180
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4187..4205
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 agcatggaac accccggaaa actgttattt gcccctaact tgctgctgga ccgcaaccag
       61 gggaagtgtg tcgaaggaat ggtggaaatt ttcgacatgc ttcttgccac ttcctcgcgt
      121 tttcgtatga tgaatttaca gggagaagag ttcgtttgtt taaagtcaat catcttactg
      181 aatagcgggg tttacacatt cttatcttcc actcttaaat cactggagga gaaagaccat
      241 attcaccgtg tcctggacaa gatcacagat actcttatcc atcttatggc gaaggcaggc
      301 ttaacacttc aacagcaaca tcagcgcctt gctcagcttc tgttgatttt aagccatatc
      361 cgtcacatgt caaacaaggg tatggaacac ctttacagta tgaagtgtaa gaacgtcgtt
      421 ccattgtatg atttgttact ggaaatgctt gacgctcacc gtctgcatgc acctggtggc
      481 ggtggctctg gtggaggcgg gagcgcagct gcatccggta ctgtcgatca aagcgatcag
      541 tcatttttag acgacgatca aattgaagct ggatacgttc tcacctgcgt tgcttaccca
      601 acttccgatt gcgtcatcga aacacacaaa gaagaagagc tttacggggg ctctggaggt
      661 tctggaggtt ctggtgggtc cgcgagtagc gaagacgtta tcaaagagtt catgcgtttc
      721 aaagttcgta tggaaggttc cgttaacggt cacgagttcg aaatcgaagg tgaaggtgaa
      781 ggtcgtccgt acgaaggtac ccagaccgct aaactgaaag ttaccaaagg tggtccgctg
      841 ccgttcgctt gggacatcct gtccccgcag ttccagtacg gttccaaagc ttacgttaaa
      901 cacccggctg acatcccgga ctacctgaaa ctgtccttcc cggaaggttt caaatgggaa
      961 cgtgttatga acttcgaaga cggtggtgtt gttaccgtta cccaggactc ctccctgcaa
     1021 gacggtgagt tcatctacaa agttaaactg cgtggtacca acttcccgtc cgacggtccg
     1081 gttatgcaga aaaaaaccat gggttgggaa gcttccaccg aacgtatgta cccggaagac
     1141 ggtgctctga aaggtgaaat caaaatgcgt ctgaaactga aagacggtgg tcactacgac
     1201 gctgaagtta aaaccaccta catggctaaa aaaccggttc agctgccggg tgcttacaaa
     1261 accgacatca aactggacat cacctcccac aacgaagact acaccatcgt tgaacagtac
     1321 gaacgtgctg aaggtcgtca ctccaccggt gcttgagaat tctgataata atctagacca
     1381 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     1441 tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg
     1501 tttataggta cccgtggatc ctctagagtc gacctgcagg catgcaagct ttttacggct
     1561 agctcagtcc taggtatagt gctagcccag ccagagaaac actctttaac agggggtagt
     1621 atgatgtctc gtttagataa aagtaaagtg attaacagcg cattagagct gcttaatgag
     1681 gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct
     1741 acattgtatt ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg
     1801 ttagataggc accatactca cttttgccct ttagaagggg aaagctggca agatttttta
     1861 cgtaataacg ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta
     1921 catttaggta cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt
     1981 ttatgccaac aaggtttttc actagagaat gcattatatg cactcagcgc agtggggcat
     2041 tttactttag gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa
     2101 acacctacta ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac
     2161 caaggtgcag agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa
     2221 caacttaaat gtgaaagtgg gtcttaaggc atcaaataaa acgaaaggct cagtcgaaag
     2281 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
     2341 cgccgcccta gacctagggc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2401 taatacggtt atccacagaa tcaggggata acgcatgcaa gaacatgtga gcaaaaggcc
     2461 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2521 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2581 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2641 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2701 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2761 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2821 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2881 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2941 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3001 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3061 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3121 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgact agtgcttgga
     3181 ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt
     3241 tctgaggtca ttactggatc tatcaacagg agtccaagcg agctcgatat caaattacgc
     3301 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
     3361 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
     3421 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
     3481 taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat
     3541 aaacccttta gggaaatagg ccaggttttc accataacac gccacatctt gcgaatatat
     3601 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
     3661 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
     3721 tttcattgcc atacgaaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
     3781 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
     3841 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
     3901 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
     3961 ttccttagct cctgaaaatc tcgataactc aaaaaatacg accggtagtg atcttatttc
     4021 attatggtga aagttggaac ctcttacggg ccggccaacg tctcattttc gccagatatc
     4081 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     4141 ccctttcgtc ttgacctcga gtccctatca gtgatagaga ttgacatccc tatcagtgat
     4201 agagatactg agcacatcag caggacgcac tgacctaaaa atccgatccc tctttagttc
     4261 tcaatcatgg caacctataa agttacactt attaacgaag ctgaaggtct aaacaaaaca
     4321 attgaagttc cagatgatca gtacatttta gatgcggctg aagaagctgg tatagactta
     4381 ccgtattctt gtcgtgctgg tgcttgctct acctgtgctg gtaaactcat atccggtgct
     4441 gcagctgggg gcggaggttc tggaggtggt gggtccaaga agaattcact ggcgttgtcg
     4501 cttacggctg accaaatggt ttctgctctg cttgacgccg aacctccgat cctgtactct
     4561 gaatacgacc cgactcgtcc tttttcagag gcgagtatga tggggctgct tacgaatctg
     4621 gcagaccgcg agttggtaca tatgattaat tgggctaaac gtgtgccggg tttcgtggac
     4681 ttgaccctgc atgaccaggt ccacctgtta gaatgcgcct ggttagagat tttaatgatc
     4741 ggacttgtct ggcgt
//
