LOCUS       sequence-259029-        4755 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 
            with a C-terminal fused mRFP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4755)
  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 4755)
  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: pBW009
FEATURES             Location/Qualifiers
     source          1..4755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..112
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(133..151,152..157,158..174,175..180,181..206)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    133..151
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    158..176
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1408..2082
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     terminator      2112..2183
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2199..2226
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2278..2312
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2347..2973
                     /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(2423..2442)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2979..3065
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3230..3818)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3310..3329)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3906..4000)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4021..4680)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4595..4614
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 aaagttggaa cctcttacgg gccggccaac gtctcatttt cgccagatat cgacgtctaa
       61 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
      121 cttgacctcg agtccctatc agtgatagag attgacatcc ctatcagtga tagagatact
      181 gagcacatca gcaggacgca ctgacctaaa aatccgatcc ctctttagtt ctcaatcatg
      241 gcaacctata aagttacact tattaacgaa gctgaaggtc taaacaaaac aattgaagtt
      301 ccagatgatc agtacatttt agatgcggct gatgctgcag ctgggggcgg aggttctgga
      361 ggtggtgggt ccaagaagaa ttcactggcg ttgtcgctta cggctgacca aatggtttct
      421 gctctgcttg acgccgaacc tccgatcctg tactctgaat acgacccgac tcgtcctttt
      481 tcagaggcga gtatgatggg gctgcttacg aatctggcag accgcgagtt ggtacatatg
      541 attaattggg ctaaacgtgt gccgggtttc gtggacttga ccctgcatga ccaggtccac
      601 ctgttagaat gcgcctggtt agagatttta atgatcggac ttgtctggcg tagcatggaa
      661 caccccggaa aactgttatt tgcccctaac ttgctgctgg accgcaacca ggggaagtgt
      721 gtcgaaggaa tggtggaaat tttcgacatg cttcttgcca cttcctcgcg ttttcgtatg
      781 atgaatttac agggagaaga gttcgtttgt ttaaagtcaa tcatcttact gaatagcggg
      841 gtttacacat tcttatcttc cactcttaaa tcactggagg agaaagacca tattcaccgt
      901 gtcctggaca agatcacaga tactcttatc catcttatgg cgaaggcagg cttaacactt
      961 caacagcaac atcagcgcct tgctcagctt ctgttgattt taagccatat ccgtcacatg
     1021 tcaaacaagg gtatggaaca cctttacagt atgaagtgta agaacgtcgt tccattgtat
     1081 gatttgttac tggaaatgct tgacgctcac cgtctgcatg cacctggtgg cggtggctct
     1141 ggtggaggcg ggagcgcagc tgcagctgaa gaagctggta tagacttacc gtattcttgt
     1201 cgtgctggtg cttgctctac ctgtgctggt aaactcatat ccggtactgt cgatcaaagc
     1261 gatcagtcat ttttagacga cgatcaaatt gaagctggat acgttctcac ctgcgttgct
     1321 tacccaactt ccgattgcgt catcgaaaca cacaaagaag aagagcttta cgggggctct
     1381 ggaggttctg gaggttctgg tgggtccgcg agtagcgaag acgttatcaa agagttcatg
     1441 cgtttcaaag ttcgtatgga aggttccgtt aacggtcacg agttcgaaat cgaaggtgaa
     1501 ggtgaaggtc gtccgtacga aggtacccag accgctaaac tgaaagttac caaaggtggt
     1561 ccgctgccgt tcgcttggga catcctgtcc ccgcagttcc agtacggttc caaagcttac
     1621 gttaaacacc cggctgacat cccggactac ctgaaactgt ccttcccgga aggtttcaaa
     1681 tgggaacgtg ttatgaactt cgaagacggt ggtgttgtta ccgttaccca ggactcctcc
     1741 ctgcaagacg gtgagttcat ctacaaagtt aaactgcgtg gtaccaactt cccgtccgac
     1801 ggtccggtta tgcagaaaaa aaccatgggt tgggaagctt ccaccgaacg tatgtacccg
     1861 gaagacggtg ctctgaaagg tgaaatcaaa atgcgtctga aactgaaaga cggtggtcac
     1921 tacgacgctg aagttaaaac cacctacatg gctaaaaaac cggttcagct gccgggtgct
     1981 tacaaaaccg acatcaaact ggacatcacc tcccacaacg aagactacac catcgttgaa
     2041 cagtacgaac gtgctgaagg tcgtcactcc accggtgctt gagaattctg ataataatct
     2101 agaccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     2161 gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt
     2221 tctgcgttta taggtacccg tggatcctct agagtcgacc tgcaggcatg caagcttttt
     2281 acggctagct cagtcctagg tatagtgcta gcccagccag agaaacactc tttaacaggg
     2341 ggtagtatga tgtctcgttt agataaaagt aaagtgatta acagcgcatt agagctgctt
     2401 aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg cccagaagct aggtgtagag
     2461 cagcctacat tgtattggca tgtaaaaaat aagcgggctt tgctcgacgc cttagccatt
     2521 gagatgttag ataggcacca tactcacttt tgccctttag aaggggaaag ctggcaagat
     2581 tttttacgta ataacgctaa aagttttaga tgtgctttac taagtcatcg cgatggagca
     2641 aaagtacatt taggtacacg gcctacagaa aaacagtatg aaactctcga aaatcaatta
     2701 gcctttttat gccaacaagg tttttcacta gagaatgcat tatatgcact cagcgcagtg
     2761 gggcatttta ctttaggttg cgtattggaa gatcaagagc atcaagtcgc taaagaagaa
     2821 agggaaacac ctactactga tagtatgccg ccattattac gacaagctat cgaattattt
     2881 gatcaccaag gtgcagagcc agccttctta ttcggccttg aattgatcat atgcggatta
     2941 gaaaaacaac ttaaatgtga aagtgggtct taaggcatca aataaaacga aaggctcagt
     3001 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     3061 caaatccgcc gccctagacc tagggcgttc ggctgcggcg agcggtatca gctcactcaa
     3121 aggcggtaat acggttatcc acagaatcag gggataacgc atgcaagaac atgtgagcaa
     3181 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3241 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3301 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3361 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3421 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3481 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3541 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3601 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3661 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3721 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3781 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3841 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgactagtg
     3901 cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
     3961 tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa
     4021 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac
     4081 atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
     4141 gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc
     4201 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt
     4261 ctcaataaac cctttaggga aataggccag gttttcacca taacacgcca catcttgcga
     4321 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt
     4381 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc
     4441 accgtctttc attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg
     4501 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat
     4561 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg
     4621 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat
     4681 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgaccg gtagtgatct
     4741 tatttcatta tggtg
//
