LOCUS       sequence-259043-        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 54 and 55.
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: pBW012
FEATURES             Location/Qualifiers
     source          1..4047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(35..53,54..59,60..76,77..82,83..108)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    35..53
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    60..78
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      1306..1377
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1393..1420
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1472..1506
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             1541..2167
                     /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(1617..1636)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2173..2259
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(2424..3012)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2504..2523)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3100..3194)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3215..3874)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3789..3808
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     join(4043..4047,1..14)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcgtatcacg aggccctttc gtcttgacct cgagtcccta tcagtgatag agattgacat
       61 ccctatcagt gatagagata ctgagcacat cagcaggacg cactgaccta aaaatccgat
      121 ccctctttag ttctcaatca tggcaaccta taaagttaca cttattaacg aagctgaagg
      181 tctaaacaaa acaattgaag ttccagatga tcagtacatt ttagatgcgg ctgaagaagc
      241 tggtatagac ttaccgtatt cttgtcgtgc tggtgcttgc tctacctgtg ctggtaaact
      301 catatctgct gcagctgggg gcggaggttc tggaggtggt gggtccaaga agaattcact
      361 ggcgttgtcg cttacggctg accaaatggt ttctgctctg cttgacgccg aacctccgat
      421 cctgtactct gaatacgacc cgactcgtcc tttttcagag gcgagtatga tggggctgct
      481 tacgaatctg gcagaccgcg agttggtaca tatgattaat tgggctaaac gtgtgccggg
      541 tttcgtggac ttgaccctgc atgaccaggt ccacctgtta gaatgcgcct ggttagagat
      601 tttaatgatc ggacttgtct ggcgtagcat ggaacacccc ggaaaactgt tatttgcccc
      661 taacttgctg ctggaccgca accaggggaa gtgtgtcgaa ggaatggtgg aaattttcga
      721 catgcttctt gccacttcct cgcgttttcg tatgatgaat ttacagggag aagagttcgt
      781 ttgtttaaag tcaatcatct tactgaatag cggggtttac acattcttat cttccactct
      841 taaatcactg gaggagaaag accatattca ccgtgtcctg gacaagatca cagatactct
      901 tatccatctt atggcgaagg caggtttaac acttcaacag caacatcagc gccttgctca
      961 gcttctgttg attttaagcc atatccgtca catgtcaaac aagggtatgg aacaccttta
     1021 cagtatgaag tgtaagaacg tcgttccatt gtatgatttg ttactggaaa tgcttgacgc
     1081 tcaccgtctg catgcacctg gtggcggtgg ctctggtgga ggcgggagcg cagctgcaat
     1141 atccggtact gtcgatcaaa gcgatcagtc atttttagac gacgatcaaa ttgaagctgg
     1201 atacgttctc acctgcgttg cttacccaac ttccgattgc gtcatcgaaa cacacaaaga
     1261 agaagagctt tactaagaat tctgataata atctagacca ggcatcaaat aaaacgaaag
     1321 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac
     1381 tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataggta cccgtggatc
     1441 ctctagagtc gacctgcagg catgcaagct ttttacggct agctcagtcc taggtatagt
     1501 gctagcccag ccagagaaac actctttaac agggggtagt atgatgtctc gtttagataa
     1561 aagtaaagtg attaacagcg cattagagct gcttaatgag gtcggaatcg aaggtttaac
     1621 aacccgtaaa ctcgcccaga agctaggtgt agagcagcct acattgtatt ggcatgtaaa
     1681 aaataagcgg gctttgctcg acgccttagc cattgagatg ttagataggc accatactca
     1741 cttttgccct ttagaagggg aaagctggca agatttttta cgtaataacg ctaaaagttt
     1801 tagatgtgct ttactaagtc atcgcgatgg agcaaaagta catttaggta cacggcctac
     1861 agaaaaacag tatgaaactc tcgaaaatca attagccttt ttatgccaac aaggtttttc
     1921 actagagaat gcattatatg cactcagcgc agtggggcat tttactttag gttgcgtatt
     1981 ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa acacctacta ctgatagtat
     2041 gccgccatta ttacgacaag ctatcgaatt atttgatcac caaggtgcag agccagcctt
     2101 cttattcggc cttgaattga tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg
     2161 gtcttaaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     2221 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgcccta gacctagggc
     2281 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     2341 tcaggggata acgcatgcaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2401 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2461 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2521 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2581 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2641 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2701 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2761 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2821 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     2881 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2941 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3001 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3061 aactcacgtt aagggatttt ggtcatgact agtgcttgga ttctcaccaa taaaaaacgc
     3121 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc
     3181 tatcaacagg agtccaagcg agctcgatat caaattacgc cccgccctgc cactcatcgc
     3241 agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat
     3301 gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg
     3361 tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac
     3421 tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg
     3481 ccaggttttc accataacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat
     3541 cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt
     3601 aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc atacgaaatt
     3661 ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct
     3721 tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg
     3781 tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat
     3841 caacggtggt atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc
     3901 tcgataactc aaaaaatacg accggtagtg atcttatttc attatggtga aagttggaac
     3961 ctcttacggg ccggccaacg tctcattttc gccagatatc gacgtctaag aaaccattat
     4021 tatcatgaca ttaacctata aaaatag
//
