LOCUS       sequence-253094-        4046 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 4046)
  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 4046)
  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..4046
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(9..103)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(124..783)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     698..717
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(784..886)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     952..970
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(991..1009,1010..1015,1016..1032,1033..1038,1039..1064)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    991..1009
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1016..1034
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      2262..2333
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2349..2376
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2428..2462
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2497..3123
                     /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(2573..2592)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      3129..3215
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3379..3967)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3459..3478)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
       61 agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgatatc
      121 aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc
      181 gacatggaag ccatcacaga cggcatgatg aacctgaatc gccagcggca tcagcacctt
      241 gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt
      301 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat
      361 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
      421 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
      481 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
      541 ctcaccgtct ttcattgcca tacgaaattc cggatgagca ttcatcaggc gggcaagaat
      601 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
      661 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
      721 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
      781 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
      841 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt ctcattttcg
      901 ccagatatcg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
      961 atcacgaggc cctttcgtct tcacctcgag tccctatcag tgatagagat tgacatccct
     1021 atcagtgata gagatactga gcacatcagc aggacgcact gacctaaaaa tccgatccct
     1081 ctttagttct caatcatggc aacctataaa gttacactta ttaacgaagc tgaaggtcta
     1141 aacaaaacaa ttgaagttcc agatgatcag tacattttag atgcggctga agaagctggt
     1201 atagacttac cgtattcttg tcgtgctggt gcttgctcta cctgtgctgg taaactcata
     1261 tctgctgcag ctgggggcgg aggttctgga ggtggtgggt ccaagaagaa ttcactggcg
     1321 ttgtcgctta cggctgacca aatggtttct gctctgcttg acgccgaacc tccgatcctg
     1381 tactctgaat acgacccgac tcgtcctttt tcagaggcga gtatgatggg gctgcttacg
     1441 aatctggcag accgcgagtt ggtacatatg attaattggg ctaaacgtgt gccgggtttc
     1501 gtggacttga ccctgcatga ccaggtccac ctgttagaat gcgcctggtt agagatttta
     1561 atgatcggac ttgtctggcg tagcatggaa caccccggaa aactgttatt tgcccctaac
     1621 ttgctgctgg accgcaacca ggggaagtgt gtcgaaggaa tggtggaaat tttcgacatg
     1681 cttcttgcca cttcctcgcg ttttcgtatg atgaatttac agggagaaga gttcgtttgt
     1741 ttaaagtcaa tcatcttact gaatagcggg gtttacacat tcttatcttc cactcttaaa
     1801 tcactggagg agaaagacca tattcaccgt gtcctggaca agatcacaga tactcttatc
     1861 catcttatgg cgaaggcagg cttaacactt caacagcaac atcagcgcct tgctcagctt
     1921 ctgttgattt taagccatat ccgtcacatg tcaaacaagg gtatggaaca cctttacagt
     1981 atgaagtgta agaacgtcgt tccattgtat gatttgttac tggaaatgct tgacgctcac
     2041 cgtctgcatg cacctggtgg cggtggctct ggtggaggcg ggagcgcagc tgcaatatcc
     2101 ggtactgtcg atcaaagcga tcagtcattt ttagacgacg atcaaattga agctggatac
     2161 gttctcacct gcgttgctta cccaacttcc gattgcgtca tcgaaacaca caaagaagaa
     2221 gagctttact aagaattctg ataataatct agaccaggca tcaaataaaa cgaaaggctc
     2281 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga
     2341 gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta taggtacccg tggatcctct
     2401 agagtcgacc tgcaggcatg caagcttttt acggctagct cagtcctagg tatagtgcta
     2461 gcccagccag agaaacactc tttaacaggg ggtagtatga tgtctcgttt agataaaagt
     2521 aaagtgatta acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc
     2581 cgtaaactcg cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat
     2641 aagcgggctt tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt
     2701 tgccctttag aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga
     2761 tgtgctttac taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa
     2821 aaacagtatg aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta
     2881 gagaatgcat tatatgcact cagcgcagtg gggcatttta ctttaggttg cgtattggaa
     2941 gatcaagagc atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg
     3001 ccattattac gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta
     3061 ttcggccttg aattgatcat atgcggatta gaaaaacaac ttaaatgtga aagtgggtct
     3121 taaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     3181 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gccctagacc taggcgttcg
     3241 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3301 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3361 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3421 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3481 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3541 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     3601 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3661 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3721 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3781 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     3841 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3901 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3961 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4021 acgttaaggg attttggtca tgacta
//
