LOCUS       sequence-259033-        4047 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 35 and 36.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4047)
  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 4047)
  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: pBW010
FEATURES             Location/Qualifiers
     source          1..4047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     69..87
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(108..126,127..132,133..149,150..155,156..181)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    108..126
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    133..151
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      1379..1450
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1466..1493
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1545..1579
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             1614..2240
                     /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(1690..1709)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2246..2332
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(2497..3085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2577..2596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3173..3267)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3288..3947)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3862..3881
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ccaacgtctc attttcgcca gatatcgacg tctaagaaac cattattatc atgacattaa
       61 cctataaaaa taggcgtatc acgaggccct ttcgtcttga cctcgagtcc ctatcagtga
      121 tagagattga catccctatc agtgatagag atactgagca catcagcagg acgcactgac
      181 ctaaaaatcc gatccctctt tagttctcaa tcatggcaac ctataaagtt acacttatta
      241 acgaagctga aggtctaaac aaaacaattg aagttccaga tgatcagtac attttagatg
      301 cggctgaaga agctggtata gatgctgcag ctgggggcgg aggttctgga ggtggtgggt
      361 ccaagaagaa ttcactggcg ttgtcgctta cggctgacca aatggtttct gctctgcttg
      421 acgccgaacc tccgatcctg tactctgaat acgacccgac tcgtcctttt tcagaggcga
      481 gtatgatggg gctgcttacg aatctggcag accgcgagtt ggtacatatg attaattggg
      541 ctaaacgtgt gccgggtttc gtggacttga ccctgcatga ccaggtccac ctgttagaat
      601 gcgcctggtt agagatttta atgatcggac ttgtctggcg tagcatggaa caccccggaa
      661 aactgttatt tgcccctaac ttgctgctgg accgcaacca ggggaagtgt gtcgaaggaa
      721 tggtggaaat tttcgacatg cttcttgcca cttcctcgcg ttttcgtatg atgaatttac
      781 agggagaaga gttcgtttgt ttaaagtcaa tcatcttact gaatagcggg gtttacacat
      841 tcttatcttc cactcttaaa tcactggagg agaaagacca tattcaccgt gtcctggaca
      901 agatcacaga tactcttatc catcttatgg cgaaggcagg cttaacactt caacagcaac
      961 atcagcgcct tgctcagctt ctgttgattt taagccatat ccgtcacatg tcaaacaagg
     1021 gtatggaaca cctttacagt atgaagtgta agaacgtcgt tccattgtat gatttgttac
     1081 tggaaatgct tgacgctcac cgtctgcatg cacctggtgg cggtggctct ggtggaggcg
     1141 ggagcgcagc tgcaatagac ttaccgtatt cttgtcgtgc tggtgcttgc tctacctgtg
     1201 ctggtaaact catatccggt actgtcgatc aaagcgatca gtcattttta gacgacgatc
     1261 aaattgaagc tggatacgtt ctcacctgcg ttgcttaccc aacttccgat tgcgtcatcg
     1321 aaacacacaa agaagaagag ctttactaag aattctgata ataatctaga ccaggcatca
     1381 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     1441 gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct gcgtttatag
     1501 gtacccgtgg atcctctaga gtcgacctgc aggcatgcaa gctttttacg gctagctcag
     1561 tcctaggtat agtgctagcc cagccagaga aacactcttt aacagggggt agtatgatgt
     1621 ctcgtttaga taaaagtaaa gtgattaaca gcgcattaga gctgcttaat gaggtcggaa
     1681 tcgaaggttt aacaacccgt aaactcgccc agaagctagg tgtagagcag cctacattgt
     1741 attggcatgt aaaaaataag cgggctttgc tcgacgcctt agccattgag atgttagata
     1801 ggcaccatac tcacttttgc cctttagaag gggaaagctg gcaagatttt ttacgtaata
     1861 acgctaaaag ttttagatgt gctttactaa gtcatcgcga tggagcaaaa gtacatttag
     1921 gtacacggcc tacagaaaaa cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc
     1981 aacaaggttt ttcactagag aatgcattat atgcactcag cgcagtgggg cattttactt
     2041 taggttgcgt attggaagat caagagcatc aagtcgctaa agaagaaagg gaaacaccta
     2101 ctactgatag tatgccgcca ttattacgac aagctatcga attatttgat caccaaggtg
     2161 cagagccagc cttcttattc ggccttgaat tgatcatatg cggattagaa aaacaactta
     2221 aatgtgaaag tgggtcttaa ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     2281 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccgcc
     2341 ctagacctag ggcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2401 gttatccaca gaatcagggg ataacgcatg caagaacatg tgagcaaaag gccagcaaaa
     2461 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2521 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2581 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2641 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2701 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2761 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2821 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2881 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     2941 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3001 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3061 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3121 tcagtggaac gaaaactcac gttaagggat tttggtcatg actagtgctt ggattctcac
     3181 caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg
     3241 tcattactgg atctatcaac aggagtccaa gcgagctcga tatcaaatta cgccccgccc
     3301 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
     3361 caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
     3421 tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
     3481 aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct
     3541 ttagggaaat aggccaggtt ttcaccataa cacgccacat cttgcgaata tatgtgtaga
     3601 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
     3661 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
     3721 gccatacgaa attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga
     3781 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg
     3841 gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc
     3901 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
     3961 gctcctgaaa atctcgataa ctcaaaaaat acgaccggta gtgatcttat ttcattatgg
     4021 tgaaagttgg aacctcttac gggccgg
//
