LOCUS       sequence-258996-        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 25 and 26.
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: pBW006
FEATURES             Location/Qualifiers
     source          1..4047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    24..42
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      1270..1341
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1357..1384
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1436..1470
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             1505..2131
                     /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(1581..1600)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2137..2223
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(2388..2976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2468..2487)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3064..3158)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3179..3838)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3753..3772
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     4007..4025
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(4046..4047,1..17,18..23,24..40,41..46,47..72)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    join(4046..4047,1..17)
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 cctatcagtg atagagattg acatccctat cagtgataga gatactgagc acatcagcag
       61 gacgcactga cctaaaaatc cgatccctct ttagttctca atcatggcaa cctataaagt
      121 tacacttatt aacgaagctg aaggtctaaa caaaacaatt gaagttccag atgatcagta
      181 cattgctgca gctgggggcg gaggttctgg aggtggtggg tccaagaaga attcactggc
      241 gttgtcgctt acggctgacc aaatggtttc tgctctgctt gacgccgaac ctccgatcct
      301 gtactctgaa tacgacccga ctcgtccttt ttcagaggcg agtatgatgg ggctgcttac
      361 gaatctggca gaccgcgagt tggtacatat gattaattgg gctaaacgtg tgccgggttt
      421 cgtggacttg accctgcatg accaggtcca cctgttagaa tgcgcctggt tagagatttt
      481 aatgatcgga cttgtctggc gtagcatgga acaccccgga aaactgttat ttgcccctaa
      541 cttgctgctg gaccgcaacc aggggaagtg tgtcgaagga atggtggaaa ttttcgacat
      601 gcttcttgcc acttcctcgc gttttcgtat gatgaattta cagggagaag agttcgtttg
      661 tttaaagtca atcatcttac tgaatagcgg ggtttacaca ttcttatctt ccactcttaa
      721 atcactggag gagaaagacc atattcaccg tgtcctggac aagatcacag atactcttat
      781 ccatcttatg gcgaaggcag gcttaacact tcaacagcaa catcagcgcc ttgctcagct
      841 tctgttgatt ttaagccata tccgtcacat gtcaaacaag ggtatggaac acctttacag
      901 tatgaagtgt aagaacgtcg ttccattgta tgatttgtta ctggaaatgc ttgacgctca
      961 ccgtctgcat gcacctggtg gcggtggctc tggtggaggc gggagcgcag ctgcatacat
     1021 tttagatgcg gctgaagaag ctggtataga cttaccgtat tcttgtcgtg ctggtgcttg
     1081 ctctacctgt gctggtaaac tcatatccgg tactgtcgat caaagcgatc agtcattttt
     1141 agacgacgat caaattgaag ctggatacgt tctcacctgc gttgcttacc caacttccga
     1201 ttgcgtcatc gaaacacaca aagaagaaga gctttactaa gaattctgat aataatctag
     1261 accaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     1321 tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc
     1381 tgcgtttata ggtacccgtg gatcctctag agtcgacctg caggcatgca agctttttac
     1441 ggctagctca gtcctaggta tagtgctagc ccagccagag aaacactctt taacaggggg
     1501 tagtatgatg tctcgtttag ataaaagtaa agtgattaac agcgcattag agctgcttaa
     1561 tgaggtcgga atcgaaggtt taacaacccg taaactcgcc cagaagctag gtgtagagca
     1621 gcctacattg tattggcatg taaaaaataa gcgggctttg ctcgacgcct tagccattga
     1681 gatgttagat aggcaccata ctcacttttg ccctttagaa ggggaaagct ggcaagattt
     1741 tttacgtaat aacgctaaaa gttttagatg tgctttacta agtcatcgcg atggagcaaa
     1801 agtacattta ggtacacggc ctacagaaaa acagtatgaa actctcgaaa atcaattagc
     1861 ctttttatgc caacaaggtt tttcactaga gaatgcatta tatgcactca gcgcagtggg
     1921 gcattttact ttaggttgcg tattggaaga tcaagagcat caagtcgcta aagaagaaag
     1981 ggaaacacct actactgata gtatgccgcc attattacga caagctatcg aattatttga
     2041 tcaccaaggt gcagagccag ccttcttatt cggccttgaa ttgatcatat gcggattaga
     2101 aaaacaactt aaatgtgaaa gtgggtctta aggcatcaaa taaaacgaaa ggctcagtcg
     2161 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     2221 aatccgccgc cctagaccta gggcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2281 gcggtaatac ggttatccac agaatcaggg gataacgcat gcaagaacat gtgagcaaaa
     2341 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2401 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2461 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2521 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2581 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2641 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2701 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2761 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2821 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2881 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2941 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3001 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gactagtgct
     3061 tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
     3121 gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt
     3181 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
     3241 ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
     3301 cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca
     3361 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct
     3421 caataaaccc tttagggaaa taggccaggt tttcaccata acacgccaca tcttgcgaat
     3481 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
     3541 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
     3601 cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa
     3661 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
     3721 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
     3781 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt
     3841 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgaccggt agtgatctta
     3901 tttcattatg gtgaaagttg gaacctctta cgggccggcc aacgtctcat tttcgccaga
     3961 tatcgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     4021 gaggcccttt cgtcttgacc tcgagtc
//
