LOCUS       sequence-253097-        4753 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 4753)
  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 4753)
  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..4753
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(133..721)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(213..232)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(809..903)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(924..1583)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1498..1517
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1584..1686)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1752..1770
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1791..1809,1810..1815,1816..1832,1833..1838,
                     1839..1864)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1791..1809
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1816..1834
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             3066..3740
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     terminator      3770..3841
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3857..3884
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        3936..3970
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             4005..4631
                     /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(4081..4100)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      4637..4723
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
       61 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
      121 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
      181 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
      241 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
      301 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
      361 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
      421 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
      481 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
      541 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
      601 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
      661 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
      721 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
      781 agggattttg gtcatgacta gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac
      841 cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga
      901 gtccaagcga gctcgatatc aaattacgcc ccgccctgcc actcatcgca gtactgttgt
      961 aattcattaa gcattctgcc gacatggaag ccatcacaga cggcatgatg aacctgaatc
     1021 gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg
     1081 gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga
     1141 ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca
     1201 ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat
     1261 tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga
     1321 acactatccc atatcaccag ctcaccgtct ttcattgcca tacgaaattc cggatgagca
     1381 ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt
     1441 acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca
     1501 actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta
     1561 tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca
     1621 aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc
     1681 cgatcaacgt ctcattttcg ccagatatcg acgtctaaga aaccattatt atcatgacat
     1741 taacctataa aaataggcgt atcacgaggc cctttcgtct tcacctcgag tccctatcag
     1801 tgatagagat tgacatccct atcagtgata gagatactga gcacatcagc aggacgcact
     1861 gacctaaaaa tccgatccct ctttagttct caatcatggc aacctataaa gttacactta
     1921 ttaacgaagc tgaaggtcta aacaaaacaa ttgaagttcc agatgatcag tacattttag
     1981 atgcggctga agaagctggt atagacttac cgtattcttg tcgtgctggt gcttgctcta
     2041 cctgtgctgg taaactcata tccggtgctg cagctggggg cggaggttct ggaggtggtg
     2101 ggtccaagaa gaattcactg gcgttgtcgc ttacggctga ccaaatggtt tctgctctgc
     2161 ttgacgccga acctccgatc ctgtactctg aatacgaccc gactcgtcct ttttcagagg
     2221 cgagtatgat ggggctgctt acgaatctgg cagaccgcga gttggtacat atgattaatt
     2281 gggctaaacg tgtgccgggt ttcgtggact tgaccctgca tgaccaggtc cacctgttag
     2341 aatgcgcctg gttagagatt ttaatgatcg gacttgtctg gcgtagcatg gaacaccccg
     2401 gaaaactgtt atttgcccct aacttgctgc tggaccgcaa ccaggggaag tgtgtcgaag
     2461 gaatggtgga aattttcgac atgcttcttg ccacttcctc gcgttttcgt atgatgaatt
     2521 tacagggaga agagttcgtt tgtttaaagt caatcatctt actgaatagc ggggtttaca
     2581 cattcttatc ttccactctt aaatcactgg aggagaaaga ccatattcac cgtgtcctgg
     2641 acaagatcac agatactctt atccatctta tggcgaaggc aggcttaaca cttcaacagc
     2701 aacatcagcg ccttgctcag cttctgttga ttttaagcca tatccgtcac atgtcaaaca
     2761 agggtatgga acacctttac agtatgaagt gtaagaacgt cgttccattg tatgatttgt
     2821 tactggaaat gcttgacgct caccgtctgc atgcacctgg tggcggtggc tctggtggag
     2881 gcgggagcgc agctgcatcc ggtactgtcg atcaaagcga tcagtcattt ttagacgacg
     2941 atcaaattga agctggatac gttctcacct gcgttgctta cccaacttcc gattgcgtca
     3001 tcgaaacaca caaagaagaa gagctttacg ggggctctgg aggttctgga ggttctggtg
     3061 ggtccgcgag tagcgaagac gttatcaaag agttcatgcg tttcaaagtt cgtatggaag
     3121 gttccgttaa cggtcacgag ttcgaaatcg aaggtgaagg tgaaggtcgt ccgtacgaag
     3181 gtacccagac cgctaaactg aaagttacca aaggtggtcc gctgccgttc gcttgggaca
     3241 tcctgtcccc gcagttccag tacggttcca aagcttacgt taaacacccg gctgacatcc
     3301 cggactacct gaaactgtcc ttcccggaag gtttcaaatg ggaacgtgtt atgaacttcg
     3361 aagacggtgg tgttgttacc gttacccagg actcctccct gcaagacggt gagttcatct
     3421 acaaagttaa actgcgtggt accaacttcc cgtccgacgg tccggttatg cagaaaaaaa
     3481 ccatgggttg ggaagcttcc accgaacgta tgtacccgga agacggtgct ctgaaaggtg
     3541 aaatcaaaat gcgtctgaaa ctgaaagacg gtggtcacta cgacgctgaa gttaaaacca
     3601 cctacatggc taaaaaaccg gttcagctgc cgggtgctta caaaaccgac atcaaactgg
     3661 acatcacctc ccacaacgaa gactacacca tcgttgaaca gtacgaacgt gctgaaggtc
     3721 gtcactccac cggtgcttaa gaattctgat aataatctag accaggcatc aaataaaacg
     3781 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     3841 ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata ggtacccgtg
     3901 gatcctctag agtcgacctg caggcatgca agctttttac ggctagctca gtcctaggta
     3961 tagtgctagc ccagccagag aaacactctt taacaggggg tagtatgatg tctcgtttag
     4021 ataaaagtaa agtgattaac agcgcattag agctgcttaa tgaggtcgga atcgaaggtt
     4081 taacaacccg taaactcgcc cagaagctag gtgtagagca gcctacattg tattggcatg
     4141 taaaaaataa gcgggctttg ctcgacgcct tagccattga gatgttagat aggcaccata
     4201 ctcacttttg ccctttagaa ggggaaagct ggcaagattt tttacgtaat aacgctaaaa
     4261 gttttagatg tgctttacta agtcatcgcg atggagcaaa agtacattta ggtacacggc
     4321 ctacagaaaa acagtatgaa actctcgaaa atcaattagc ctttttatgc caacaaggtt
     4381 tttcactaga gaatgcatta tatgcactca gcgcagtggg gcattttact ttaggttgcg
     4441 tattggaaga tcaagagcat caagtcgcta aagaagaaag ggaaacacct actactgata
     4501 gtatgccgcc attattacga caagctatcg aattatttga tcaccaaggt gcagagccag
     4561 ccttcttatt cggccttgaa ttgatcatat gcggattaga aaaacaactt aaatgtgaaa
     4621 gtgggtctta aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     4681 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgc cctagaccta
     4741 ggcgttcggc tgc
//
