LOCUS       sequence-259071-        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 54 and 55 
            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: pBW013
FEATURES             Location/Qualifiers
     source          1..4755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     84..102
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(123..141,142..147,148..164,165..170,171..196)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    123..141
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    148..166
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1398..2072
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     terminator      2102..2173
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2189..2216
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2268..2302
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2337..2963
                     /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(2413..2432)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2969..3055
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3220..3808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3300..3319)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3896..3990)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4011..4670)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4585..4604
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 cctcttacgg gccggccaac gtctcatttt cgccagatat cgacgtctaa gaaaccatta
       61 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttgacctcg
      121 agtccctatc agtgatagag attgacatcc ctatcagtga tagagatact gagcacatca
      181 gcaggacgca ctgacctaaa aatccgatcc ctctttagtt ctcaatcatg gcaacctata
      241 aagttacact tattaacgaa gctgaaggtc taaacaaaac aattgaagtt ccagatgatc
      301 agtacatttt agatgcggct gaagaagctg gtatagactt accgtattct tgtcgtgctg
      361 gtgcttgctc tacctgtgct ggtaaactca tatctgctgc agctgggggc ggaggttctg
      421 gaggtggtgg gtccaagaag aattcactgg cgttgtcgct tacggctgac caaatggttt
      481 ctgctctgct tgacgccgaa cctccgatcc tgtactctga atacgacccg actcgtcctt
      541 tttcagaggc gagtatgatg gggctgctta cgaatctggc agaccgcgag ttggtacata
      601 tgattaattg ggctaaacgt gtgccgggtt tcgtggactt gaccctgcat gaccaggtcc
      661 acctgttaga atgcgcctgg ttagagattt taatgatcgg acttgtctgg cgtagcatgg
      721 aacaccccgg aaaactgtta tttgccccta acttgctgct ggaccgcaac caggggaagt
      781 gtgtcgaagg aatggtggaa attttcgaca tgcttcttgc cacttcctcg cgttttcgta
      841 tgatgaattt acagggagaa gagttcgttt gtttaaagtc aatcatctta ctgaatagcg
      901 gggtttacac attcttatct tccactctta aatcactgga ggagaaagac catattcacc
      961 gtgtcctgga caagatcaca gatactctta tccatcttat ggcgaaggca ggtttaacac
     1021 ttcaacagca acatcagcgc cttgctcagc ttctgttgat tttaagccat atccgtcaca
     1081 tgtcaaacaa gggtatggaa cacctttaca gtatgaagtg taagaacgtc gttccattgt
     1141 atgatttgtt actggaaatg cttgacgctc accgtctgca tgcacctggt ggcggtggct
     1201 ctggtggagg cgggagcgca gctgcaatat ccggtactgt cgatcaaagc gatcagtcat
     1261 ttttagacga cgatcaaatt gaagctggat acgttctcac ctgcgttgct tacccaactt
     1321 ccgattgcgt catcgaaaca cacaaagaag aagagcttta cgggggctct ggaggttctg
     1381 gaggttctgg tgggtccgcg agtagcgaag acgttatcaa agagttcatg cgtttcaaag
     1441 ttcgtatgga aggttccgtt aacggtcacg agttcgaaat cgaaggtgaa ggtgaaggtc
     1501 gtccgtacga aggtacccag accgctaaac tgaaagttac caaaggtggt ccgctgccgt
     1561 tcgcttggga catcctgtcc ccgcagttcc agtacggttc caaagcttac gttaaacacc
     1621 cggctgacat cccggactac ctgaaactgt ccttcccgga aggtttcaaa tgggaacgtg
     1681 ttatgaactt cgaagacggt ggtgttgtta ccgttaccca ggactcctcc ctgcaagacg
     1741 gtgagttcat ctacaaagtt aaactgcgtg gtaccaactt cccgtccgac ggtccggtta
     1801 tgcagaaaaa aaccatgggt tgggaagctt ccaccgaacg tatgtacccg gaagacggtg
     1861 ctctgaaagg tgaaatcaaa atgcgtctga aactgaaaga cggtggtcac tacgacgctg
     1921 aagttaaaac cacctacatg gctaaaaaac cggttcagct gccgggtgct tacaaaaccg
     1981 acatcaaact ggacatcacc tcccacaacg aagactacac catcgttgaa cagtacgaac
     2041 gtgctgaagg tcgtcactcc accggtgctt gagaattctg ataataatct agaccaggca
     2101 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     2161 ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta
     2221 taggtacccg tggatcctct agagtcgacc tgcaggcatg caagcttttt acggctagct
     2281 cagtcctagg tatagtgcta gcccagccag agaaacactc tttaacaggg ggtagtatga
     2341 tgtctcgttt agataaaagt aaagtgatta acagcgcatt agagctgctt aatgaggtcg
     2401 gaatcgaagg tttaacaacc cgtaaactcg cccagaagct aggtgtagag cagcctacat
     2461 tgtattggca tgtaaaaaat aagcgggctt tgctcgacgc cttagccatt gagatgttag
     2521 ataggcacca tactcacttt tgccctttag aaggggaaag ctggcaagat tttttacgta
     2581 ataacgctaa aagttttaga tgtgctttac taagtcatcg cgatggagca aaagtacatt
     2641 taggtacacg gcctacagaa aaacagtatg aaactctcga aaatcaatta gcctttttat
     2701 gccaacaagg tttttcacta gagaatgcat tatatgcact cagcgcagtg gggcatttta
     2761 ctttaggttg cgtattggaa gatcaagagc atcaagtcgc taaagaagaa agggaaacac
     2821 ctactactga tagtatgccg ccattattac gacaagctat cgaattattt gatcaccaag
     2881 gtgcagagcc agccttctta ttcggccttg aattgatcat atgcggatta gaaaaacaac
     2941 ttaaatgtga aagtgggtct taaggcatca aataaaacga aaggctcagt cgaaagactg
     3001 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     3061 gccctagacc tagggcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3121 acggttatcc acagaatcag gggataacgc atgcaagaac atgtgagcaa aaggccagca
     3181 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3241 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3301 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3361 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3421 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3481 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3541 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3601 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3661 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3721 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3781 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3841 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgactagtg cttggattct
     3901 caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg
     3961 aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa ttacgccccg
     4021 ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca
     4081 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
     4141 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa
     4201 tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac
     4261 cctttaggga aataggccag gttttcacca taacacgcca catcttgcga atatatgtgt
     4321 agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc
     4381 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc
     4441 attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc
     4501 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga
     4561 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga
     4621 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc
     4681 ttagctcctg aaaatctcga taactcaaaa aatacgaccg gtagtgatct tatttcatta
     4741 tggtgaaagt tggaa
//
