LOCUS       sequence-259073-        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 55 and 56.
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: pBW014
FEATURES             Location/Qualifiers
     source          1..4047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     109..127
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(148..166,167..172,173..189,190..195,196..221)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    148..166
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    173..191
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      1419..1490
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1506..1533
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1585..1619
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             1654..2280
                     /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(1730..1749)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2286..2372
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(2537..3125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2617..2636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3213..3307)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3328..3987)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3902..3921
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tatttcatta tggtgaaagt tggaacctct tacgggccgg ccaacgtctc attttcgcca
       61 gatatcgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
      121 acgaggccct ttcgtcttga cctcgagtcc ctatcagtga tagagattga catccctatc
      181 agtgatagag atactgagca catcagcagg acgcactgac ctaaaaatcc gatccctctt
      241 tagttctcaa tcatggcaac ctataaagtt acacttatta acgaagctga aggtctaaac
      301 aaaacaattg aagttccaga tgatcagtac attttagatg cggctgaaga agctggtata
      361 gacttaccgt attcttgtcg tgctggtgct tgctctacct gtgctggtaa actcatatcc
      421 ggtgctgcag ctgggggcgg aggttctgga ggtggtgggt ccaagaagaa ttcactggcg
      481 ttgtcgctta cggctgacca aatggtttct gctctgcttg acgccgaacc tccgatcctg
      541 tactctgaat acgacccgac tcgtcctttt tcagaggcga gtatgatggg gctgcttacg
      601 aatctggcag accgcgagtt ggtacatatg attaattggg ctaaacgtgt gccgggtttc
      661 gtggacttga ccctgcatga ccaggtccac ctgttagaat gcgcctggtt agagatttta
      721 atgatcggac ttgtctggcg tagcatggaa caccccggaa aactgttatt tgcccctaac
      781 ttgctgctgg accgcaacca ggggaagtgt gtcgaaggaa tggtggaaat tttcgacatg
      841 cttcttgcca cttcctcgcg ttttcgtatg atgaatttac agggagaaga gttcgtttgt
      901 ttaaagtcaa tcatcttact gaatagcggg gtttacacat tcttatcttc cactcttaaa
      961 tcactggagg agaaagacca tattcaccgt gtcctggaca agatcacaga tactcttatc
     1021 catcttatgg cgaaggcagg cttaacactt caacagcaac atcagcgcct tgctcagctt
     1081 ctgttgattt taagccatat ccgtcacatg tcaaacaagg gtatggaaca cctttacagt
     1141 atgaagtgta agaacgtcgt tccattgtat gatttgttac tggaaatgct tgacgctcac
     1201 cgtctgcatg cacctggtgg cggtggctct ggtggaggcg ggagcgcagc tgcatccggt
     1261 actgtcgatc aaagcgatca gtcattttta gacgacgatc aaattgaagc tggatacgtt
     1321 ctcacctgcg ttgcttaccc aacttccgat tgcgtcatcg aaacacacaa agaagaagag
     1381 ctttactaag aattctgata ataatctaga ccaggcatca aataaaacga aaggctcagt
     1441 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc
     1501 acactggctc accttcgggt gggcctttct gcgtttatag gtacccgtgg atcctctaga
     1561 gtcgacctgc aggcatgcaa gctttttacg gctagctcag tcctaggtat agtgctagcc
     1621 cagccagaga aacactcttt aacagggggt agtatgatgt ctcgtttaga taaaagtaaa
     1681 gtgattaaca gcgcattaga gctgcttaat gaggtcggaa tcgaaggttt aacaacccgt
     1741 aaactcgccc agaagctagg tgtagagcag cctacattgt attggcatgt aaaaaataag
     1801 cgggctttgc tcgacgcctt agccattgag atgttagata ggcaccatac tcacttttgc
     1861 cctttagaag gggaaagctg gcaagatttt ttacgtaata acgctaaaag ttttagatgt
     1921 gctttactaa gtcatcgcga tggagcaaaa gtacatttag gtacacggcc tacagaaaaa
     1981 cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc aacaaggttt ttcactagag
     2041 aatgcattat atgcactcag cgcagtgggg cattttactt taggttgcgt attggaagat
     2101 caagagcatc aagtcgctaa agaagaaagg gaaacaccta ctactgatag tatgccgcca
     2161 ttattacgac aagctatcga attatttgat caccaaggtg cagagccagc cttcttattc
     2221 ggccttgaat tgatcatatg cggattagaa aaacaactta aatgtgaaag tgggtcttaa
     2281 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     2341 tgtcggtgaa cgctctcctg agtaggacaa atccgccgcc ctagacctag ggcgttcggc
     2401 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2461 ataacgcatg caagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2521 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2581 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2641 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2701 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2761 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2821 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2881 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2941 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     3001 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3061 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3121 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3181 gttaagggat tttggtcatg actagtgctt ggattctcac caataaaaaa cgcccggcgg
     3241 caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac
     3301 aggagtccaa gcgagctcga tatcaaatta cgccccgccc tgccactcat cgcagtactg
     3361 ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg
     3421 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac
     3481 gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca
     3541 gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt
     3601 ttcaccataa cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg
     3661 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg
     3721 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa attccggatg
     3781 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt
     3841 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg
     3901 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt
     3961 ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa
     4021 ctcaaaaaat acgaccggta gtgatct
//
