LOCUS       sequence-259039-        4755 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Vector with aTc inducible M. laminosus Fd1 mutant with the estrogen 
            recepor-ligand binding domain fused between amino acids 35 and 36 
            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 Apr 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: pBW011
FEATURES             Location/Qualifiers
     source          1..4755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             966..1640
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     terminator      1670..1741
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1757..1784
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1836..1870
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             1905..2531
                     /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(1981..2000)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2537..2623
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(2788..3376)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2868..2887)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3464..3558)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3579..4238)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4153..4172
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     4407..4425
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(4446..4464,4465..4470,4471..4487,4488..4493,
                     4494..4519)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    4446..4464
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4471..4489
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 ttgacgccga acctccgatc ctgtactctg aatacgaccc gactcgtcct ttttcagagg
       61 cgagtatgat ggggctgctt acgaatctgg cagaccgcga gttggtacat atgattaatt
      121 gggctaaacg tgtgccgggt ttcgtggact tgaccctgca tgaccaggtc cacctgttag
      181 aatgcgcctg gttagagatt ttaatgatcg gacttgtctg gcgtagcatg gaacaccccg
      241 gaaaactgtt atttgcccct aacttgctgc tggaccgcaa ccaggggaag tgtgtcgaag
      301 gaatggtgga aattttcgac atgcttcttg ccacttcctc gcgttttcgt atgatgaatt
      361 tacagggaga agagttcgtt tgtttaaagt caatcatctt actgaatagc ggggtttaca
      421 cattcttatc ttccactctt aaatcactgg aggagaaaga ccatattcac cgtgtcctgg
      481 acaagatcac agatactctt atccatctta tggcgaaggc aggcttaaca cttcaacagc
      541 aacatcagcg ccttgctcag cttctgttga ttttaagcca tatccgtcac atgtcaaaca
      601 agggtatgga acacctttac agtatgaagt gtaagaacgt cgttccattg tatgatttgt
      661 tactggaaat gcttgacgct caccgtctgc atgcacctgg tggcggtggc tctggtggag
      721 gcgggagcgc agctgcaata gacttaccgt attcttgtcg tgctggtgct tgctctacct
      781 gtgctggtaa actcatatcc ggtactgtcg atcaaagcga tcagtcattt ttagacgacg
      841 atcaaattga agctggatac gttctcacct gcgttgctta cccaacttcc gattgcgtca
      901 tcgaaacaca caaagaagaa gagctttacg ggggctctgg aggttctgga ggttctggtg
      961 ggtccgcgag tagcgaagac gttatcaaag agttcatgcg tttcaaagtt cgtatggaag
     1021 gttccgttaa cggtcacgag ttcgaaatcg aaggtgaagg tgaaggtcgt ccgtacgaag
     1081 gtacccagac cgctaaactg aaagttacca aaggtggtcc gctgccgttc gcttgggaca
     1141 tcctgtcccc gcagttccag tacggttcca aagcttacgt taaacacccg gctgacatcc
     1201 cggactacct gaaactgtcc ttcccggaag gtttcaaatg ggaacgtgtt atgaacttcg
     1261 aagacggtgg tgttgttacc gttacccagg actcctccct gcaagacggt gagttcatct
     1321 acaaagttaa actgcgtggt accaacttcc cgtccgacgg tccggttatg cagaaaaaaa
     1381 ccatgggttg ggaagcttcc accgaacgta tgtacccgga agacggtgct ctgaaaggtg
     1441 aaatcaaaat gcgtctgaaa ctgaaagacg gtggtcacta cgacgctgaa gttaaaacca
     1501 cctacatggc taaaaaaccg gttcagctgc cgggtgctta caaaaccgac atcaaactgg
     1561 acatcacctc ccacaacgaa gactacacca tcgttgaaca gtacgaacgt gctgaaggtc
     1621 gtcactccac cggtgcttga gaattctgat aataatctag accaggcatc aaataaaacg
     1681 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     1741 ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata ggtacccgtg
     1801 gatcctctag agtcgacctg caggcatgca agctttttac ggctagctca gtcctaggta
     1861 tagtgctagc ccagccagag aaacactctt taacaggggg tagtatgatg tctcgtttag
     1921 ataaaagtaa agtgattaac agcgcattag agctgcttaa tgaggtcgga atcgaaggtt
     1981 taacaacccg taaactcgcc cagaagctag gtgtagagca gcctacattg tattggcatg
     2041 taaaaaataa gcgggctttg ctcgacgcct tagccattga gatgttagat aggcaccata
     2101 ctcacttttg ccctttagaa ggggaaagct ggcaagattt tttacgtaat aacgctaaaa
     2161 gttttagatg tgctttacta agtcatcgcg atggagcaaa agtacattta ggtacacggc
     2221 ctacagaaaa acagtatgaa actctcgaaa atcaattagc ctttttatgc caacaaggtt
     2281 tttcactaga gaatgcatta tatgcactca gcgcagtggg gcattttact ttaggttgcg
     2341 tattggaaga tcaagagcat caagtcgcta aagaagaaag ggaaacacct actactgata
     2401 gtatgccgcc attattacga caagctatcg aattatttga tcaccaaggt gcagagccag
     2461 ccttcttatt cggccttgaa ttgatcatat gcggattaga aaaacaactt aaatgtgaaa
     2521 gtgggtctta aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     2581 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgc cctagaccta
     2641 gggcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2701 agaatcaggg gataacgcat gcaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2761 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2821 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2881 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2941 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3001 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3061 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3121 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3181 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     3241 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3301 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3361 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3421 cgaaaactca cgttaaggga ttttggtcat gactagtgct tggattctca ccaataaaaa
     3481 acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg
     3541 gatctatcaa caggagtcca agcgagctcg atatcaaatt acgccccgcc ctgccactca
     3601 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca
     3661 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
     3721 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
     3781 aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa
     3841 taggccaggt tttcaccata acacgccaca tcttgcgaat atatgtgtag aaactgccgg
     3901 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
     3961 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacga
     4021 aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
     4081 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
     4141 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
     4201 atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa
     4261 aatctcgata actcaaaaaa tacgaccggt agtgatctta tttcattatg gtgaaagttg
     4321 gaacctctta cgggccggcc aacgtctcat tttcgccaga tatcgacgtc taagaaacca
     4381 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttgacc
     4441 tcgagtccct atcagtgata gagattgaca tccctatcag tgatagagat actgagcaca
     4501 tcagcaggac gcactgacct aaaaatccga tccctcttta gttctcaatc atggcaacct
     4561 ataaagttac acttattaac gaagctgaag gtctaaacaa aacaattgaa gttccagatg
     4621 atcagtacat tttagatgcg gctgaagaag ctggtataga tgctgcagct gggggcggag
     4681 gttctggagg tggtgggtcc aagaagaatt cactggcgtt gtcgcttacg gctgaccaaa
     4741 tggtttctgc tctgc
//
