LOCUS       sequence-253089-        4754 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 
            with a C-terminal fused mRFP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4754)
  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 4754)
  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: pBW007
FEATURES             Location/Qualifiers
     source          1..4754
                     /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"
     CDS             2266..2940
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     terminator      2970..3041
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3057..3084
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        3136..3170
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             3205..3831
                     /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(3281..3300)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      3837..3923
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(4087..4675)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4167..4186)
                     /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 tgacctcgag tccctatcag tgatagagat tgacatccct
     1021 atcagtgata gagatactga gcacatcagc aggacgcact gacctaaaaa tccgatccct
     1081 ctttagttct caatcatggc aacctataaa gttacactta ttaacgaagc tgaaggtcta
     1141 aacaaaacaa ttgaagttcc agatgatcag tacattgctg cagctggggg cggaggttct
     1201 ggaggtggtg ggtccaagaa gaattcactg gcgttgtcgc ttacggctga ccaaatggtt
     1261 tctgctctgc ttgacgccga acctccgatc ctgtactctg aatacgaccc gactcgtcct
     1321 ttttcagagg cgagtatgat ggggctgctt acgaatctgg cagaccgcga gttggtacat
     1381 atgattaatt gggctaaacg tgtgccgggt ttcgtggact tgaccctgca tgaccaggtc
     1441 cacctgttag aatgcgcctg gttagagatt ttaatgatcg gacttgtctg gcgtagcatg
     1501 gaacaccccg gaaaactgtt atttgcccct aacttgctgc tggaccgcaa ccaggggaag
     1561 tgtgtcgaag gaatggtgga aattttcgac atgcttcttg ccacttcctc gcgttttcgt
     1621 atgatgaatt tacagggaga agagttcgtt tgtttaaagt caatcatctt actgaatagc
     1681 ggggtttaca cattcttatc ttccactctt aaatcactgg aggagaaaga ccatattcac
     1741 cgtgtcctgg acaagatcac agatactctt atccatctta tggcgaaggc aggcttaaca
     1801 cttcaacagc aacatcagcg ccttgctcag cttctgttga ttttaagcca tatccgtcac
     1861 atgtcaaaca agggtatgga acacctttac agtatgaagt gtaagaacgt cgttccattg
     1921 tatgatttgt tactggaaat gcttgacgct caccgtctgc atgcacctgg tggcggtggc
     1981 tctggtggag gcgggagcgc agctgcatac attttagatg cggctgaaga agctggtata
     2041 gacttaccgt attcttgtcg tgctggtgct tgctctacct gtgctggtaa actcatatcc
     2101 ggtactgtcg atcaaagcga tcagtcattt ttagacgacg atcaaattga agctggatac
     2161 gttctcacct gcgttgctta cccaacttcc gattgcgtca tcgaaacaca caaagaagaa
     2221 gagctttacg ggggctctgg aggttctgga ggttctggtg ggtccgcgag tagcgaagac
     2281 gttatcaaag agttcatgcg tttcaaagtt cgtatggaag gttccgttaa cggtcacgag
     2341 ttcgaaatcg aaggtgaagg tgaaggtcgt ccgtacgaag gtacccagac cgctaaactg
     2401 aaagttacca aaggtggtcc gctgccgttc gcttgggaca tcctgtcccc gcagttccag
     2461 tacggttcca aagcttacgt taaacacccg gctgacatcc cggactacct gaaactgtcc
     2521 ttcccggaag gtttcaaatg ggaacgtgtt atgaacttcg aagacggtgg tgttgttacc
     2581 gttacccagg actcctccct gcaagacggt gagttcatct acaaagttaa actgcgtggt
     2641 accaacttcc cgtccgacgg tccggttatg cagaaaaaaa ccatgggttg ggaagcttcc
     2701 accgaacgta tgtacccgga agacggtgct ctgaaaggtg aaatcaaaat gcgtctgaaa
     2761 ctgaaagacg gtggtcacta cgacgctgaa gttaaaacca cctacatggc taaaaaaccg
     2821 gttcagctgc cgggtgctta caaaaccgac atcaaactgg acatcacctc ccacaacgaa
     2881 gactacacca tcgttgaaca gtacgaacgt gctgaaggtc gtcactccac cggtgcttga
     2941 gaattctgat aataatctag accaggcatc aaataaaacg aaaggctcag tcgaaagact
     3001 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     3061 caccttcggg tgggcctttc tgcgtttata ggtacccgtg gatcctctag agtcgacctg
     3121 caggcatgca agctttttac ggctagctca gtcctaggta tagtgctagc ccagccagag
     3181 aaacactctt taacaggggg tagtatgatg tctcgtttag ataaaagtaa agtgattaac
     3241 agcgcattag agctgcttaa tgaggtcgga atcgaaggtt taacaacccg taaactcgcc
     3301 cagaagctag gtgtagagca gcctacattg tattggcatg taaaaaataa gcgggctttg
     3361 ctcgacgcct tagccattga gatgttagat aggcaccata ctcacttttg ccctttagaa
     3421 ggggaaagct ggcaagattt tttacgtaat aacgctaaaa gttttagatg tgctttacta
     3481 agtcatcgcg atggagcaaa agtacattta ggtacacggc ctacagaaaa acagtatgaa
     3541 actctcgaaa atcaattagc ctttttatgc caacaaggtt tttcactaga gaatgcatta
     3601 tatgcactca gcgcagtggg gcattttact ttaggttgcg tattggaaga tcaagagcat
     3661 caagtcgcta aagaagaaag ggaaacacct actactgata gtatgccgcc attattacga
     3721 caagctatcg aattatttga tcaccaaggt gcagagccag ccttcttatt cggccttgaa
     3781 ttgatcatat gcggattaga aaaacaactt aaatgtgaaa gtgggtctta aggcatcaaa
     3841 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     3901 acgctctcct gagtaggaca aatccgccgc cctagaccta ggcgttcggc tgcggcgagc
     3961 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4021 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4081 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4141 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4201 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4261 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4321 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4381 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4441 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4501 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     4561 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4621 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4681 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4741 tttggtcatg acta
//
