LOCUS       sequence-258986-        3241 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector with aTc inducible M. laminosus Fd1 flanked by BbsI 
            restriction sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3241)
  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 3241)
  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: pBW003
FEATURES             Location/Qualifiers
     source          1..3241
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     35..53
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(74..92,93..98,99..115,116..121,122..147)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    74..92
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    99..117
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      539..610
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      626..653
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        705..739
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             774..1400
                     /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(850..869)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      1406..1492
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1657..2245)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1737..1756)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2333..2427)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2448..3107)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3022..3041
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
       61 tcttgacctc gagtccctat cagtgataga gattgacatc cctatcagtg atagagatac
      121 tgagcacatc agcaggacgc actgacctaa aaatccgatc cctctttagt tctcaatcgc
      181 ggccgcgaag acatcatggc aacctataaa gttacactta ttaacgaagc tgaaggtcta
      241 aacaaaacaa ttgaagttcc agatgatcag tacattttag atgcggctga agaagctggt
      301 atagacttac cgtattcttg tcgtgctggt gcttgctcta cctgtgctgg taaactcata
      361 tccggtactg tcgatcaaag cgatcagtca tttttagacg acgatcaaat tgaagctgga
      421 tacgttctca cctgcgttgc ttacccaact tccgattgcg tcatcgaaac acacaaagaa
      481 gaagagcttt actaagtctt cgcggccgcg aattctgata ataatctaga ccaggcatca
      541 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
      601 gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct gcgtttatag
      661 gtacccgtgg atcctctaga gtcgacctgc aggcatgcaa gctttttacg gctagctcag
      721 tcctaggtat agtgctagcc cagccagaga aacactcttt aacagggggt agtatgatgt
      781 ctcgtttaga taaaagtaaa gtgattaaca gcgcattaga gctgcttaat gaggtcggaa
      841 tcgaaggttt aacaacccgt aaactcgccc agaagctagg tgtagagcag cctacattgt
      901 attggcatgt aaaaaataag cgggctttgc tcgacgcctt agccattgag atgttagata
      961 ggcaccatac tcacttttgc cctttagaag gggaaagctg gcaagatttt ttacgtaata
     1021 acgctaaaag ttttagatgt gctttactaa gtcatcgcga tggagcaaaa gtacatttag
     1081 gtacacggcc tacagaaaaa cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc
     1141 aacaaggttt ttcactagag aatgcattat atgcactcag cgcagtgggg cattttactt
     1201 taggttgcgt attggaagat caagagcatc aagtcgctaa agaagaaagg gaaacaccta
     1261 ctactgatag tatgccgcca ttattacgac aagctatcga attatttgat caccaaggtg
     1321 cagagccagc cttcttattc ggccttgaat tgatcatatg cggattagaa aaacaactta
     1381 aatgtgaaag tgggtcttaa ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     1441 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccgcc
     1501 ctagacctag ggcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1561 gttatccaca gaatcagggg ataacgcatg caagaacatg tgagcaaaag gccagcaaaa
     1621 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1681 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1741 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1801 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1861 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1921 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1981 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2041 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     2101 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2161 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2221 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2281 tcagtggaac gaaaactcac gttaagggat tttggtcatg actagtgctt ggattctcac
     2341 caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg
     2401 tcattactgg atctatcaac aggagtccaa gcgagctcga tatcaaatta cgccccgccc
     2461 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
     2521 caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
     2581 tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
     2641 aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct
     2701 ttagggaaat aggccaggtt ttcaccataa cacgccacat cttgcgaata tatgtgtaga
     2761 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
     2821 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
     2881 gccatacgaa attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga
     2941 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg
     3001 gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc
     3061 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
     3121 gctcctgaaa atctcgataa ctcaaaaaat acgaccggta gtgatcttat ttcattatgg
     3181 tgaaagttgg aacctcttac gggccggcca acgtctcatt ttcgccagat atcgacgtct
     3241 a
//
