LOCUS       sequence-259026-        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 30 and 31.
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: pBW008
FEATURES             Location/Qualifiers
     source          1..4047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     83..101
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(122..140,141..146,147..163,164..169,170..195)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    122..140
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    147..165
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      1393..1464
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1480..1507
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1559..1593
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             1628..2254
                     /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(1704..1723)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2260..2346
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(2511..3099)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2591..2610)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3187..3281)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3302..3961)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3876..3895
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ctcttacggg ccggccaacg tctcattttc gccagatatc gacgtctaag aaaccattat
       61 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttgacctcga
      121 gtccctatca gtgatagaga ttgacatccc tatcagtgat agagatactg agcacatcag
      181 caggacgcac tgacctaaaa atccgatccc tctttagttc tcaatcatgg caacctataa
      241 agttacactt attaacgaag ctgaaggtct aaacaaaaca attgaagttc cagatgatca
      301 gtacatttta gatgcggctg atgctgcagc tgggggcgga ggttctggag gtggtgggtc
      361 caagaagaat tcactggcgt tgtcgcttac ggctgaccaa atggtttctg ctctgcttga
      421 cgccgaacct ccgatcctgt actctgaata cgacccgact cgtccttttt cagaggcgag
      481 tatgatgggg ctgcttacga atctggcaga ccgcgagttg gtacatatga ttaattgggc
      541 taaacgtgtg ccgggtttcg tggacttgac cctgcatgac caggtccacc tgttagaatg
      601 cgcctggtta gagattttaa tgatcggact tgtctggcgt agcatggaac accccggaaa
      661 actgttattt gcccctaact tgctgctgga ccgcaaccag gggaagtgtg tcgaaggaat
      721 ggtggaaatt ttcgacatgc ttcttgccac ttcctcgcgt tttcgtatga tgaatttaca
      781 gggagaagag ttcgtttgtt taaagtcaat catcttactg aatagcgggg tttacacatt
      841 cttatcttcc actcttaaat cactggagga gaaagaccat attcaccgtg tcctggacaa
      901 gatcacagat actcttatcc atcttatggc gaaggcaggc ttaacacttc aacagcaaca
      961 tcagcgcctt gctcagcttc tgttgatttt aagccatatc cgtcacatgt caaacaaggg
     1021 tatggaacac ctttacagta tgaagtgtaa gaacgtcgtt ccattgtatg atttgttact
     1081 ggaaatgctt gacgctcacc gtctgcatgc acctggtggc ggtggctctg gtggaggcgg
     1141 gagcgcagct gcagctgaag aagctggtat agacttaccg tattcttgtc gtgctggtgc
     1201 ttgctctacc tgtgctggta aactcatatc cggtactgtc gatcaaagcg atcagtcatt
     1261 tttagacgac gatcaaattg aagctggata cgttctcacc tgcgttgctt acccaacttc
     1321 cgattgcgtc atcgaaacac acaaagaaga agagctttac taagaattct gataataatc
     1381 tagaccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     1441 tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct
     1501 ttctgcgttt ataggtaccc gtggatcctc tagagtcgac ctgcaggcat gcaagctttt
     1561 tacggctagc tcagtcctag gtatagtgct agcccagcca gagaaacact ctttaacagg
     1621 gggtagtatg atgtctcgtt tagataaaag taaagtgatt aacagcgcat tagagctgct
     1681 taatgaggtc ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc taggtgtaga
     1741 gcagcctaca ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg ccttagccat
     1801 tgagatgtta gataggcacc atactcactt ttgcccttta gaaggggaaa gctggcaaga
     1861 ttttttacgt aataacgcta aaagttttag atgtgcttta ctaagtcatc gcgatggagc
     1921 aaaagtacat ttaggtacac ggcctacaga aaaacagtat gaaactctcg aaaatcaatt
     1981 agccttttta tgccaacaag gtttttcact agagaatgca ttatatgcac tcagcgcagt
     2041 ggggcatttt actttaggtt gcgtattgga agatcaagag catcaagtcg ctaaagaaga
     2101 aagggaaaca cctactactg atagtatgcc gccattatta cgacaagcta tcgaattatt
     2161 tgatcaccaa ggtgcagagc cagccttctt attcggcctt gaattgatca tatgcggatt
     2221 agaaaaacaa cttaaatgtg aaagtgggtc ttaaggcatc aaataaaacg aaaggctcag
     2281 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     2341 acaaatccgc cgccctagac ctagggcgtt cggctgcggc gagcggtatc agctcactca
     2401 aaggcggtaa tacggttatc cacagaatca ggggataacg catgcaagaa catgtgagca
     2461 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2521 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2581 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2641 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2701 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2761 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2821 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2881 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2941 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3001 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3061 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3121 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgactagt
     3181 gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag
     3241 atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc tcgatatcaa
     3301 attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga
     3361 catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt
     3421 cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg
     3481 ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat
     3541 tctcaataaa ccctttaggg aaataggcca ggttttcacc ataacacgcc acatcttgcg
     3601 aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg
     3661 tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct
     3721 caccgtcttt cattgccata cgaaattccg gatgagcatt catcaggcgg gcaagaatgt
     3781 gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa
     3841 tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat
     3901 gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca
     3961 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgacc ggtagtgatc
     4021 ttatttcatt atggtgaaag ttggaac
//
