LOCUS       sequence-258983-        3210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector with aTc inducible M. laminosus Fd1 without BbsI restriction 
            sites in the vector backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3210)
  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 3210)
  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: pBW002
FEATURES             Location/Qualifiers
     source          1..3210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     64..82
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(103..121,122..127,128..144,145..150,151..176)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    103..121
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    128..146
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      537..608
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      624..651
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        703..737
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             772..1398
                     /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(848..867)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      1404..1490
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1655..2243)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1735..1754)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2331..2425)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2446..3105)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3020..3039
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gtctcatttt cgccagatat cgacgtctaa gaaaccatta ttatcatgac attaacctat
       61 aaaaataggc gtatcacgag gccctttcgt cttgacctcg agtccctatc agtgatagag
      121 attgacatcc ctatcagtga tagagatact gagcacatca gcaggacgca ctgacctaaa
      181 aatccgatcc ctctttagtt ctcaatcatg gcaacctata aagttacact tattaacgaa
      241 gctgaaggtc taaacaaaac aattgaagtt ccagatgatc agtacatttt agatgcggct
      301 gaagaagctg gtatagactt accgtattct tgtcgtgctg gtgcttgctc tacctgtgct
      361 ggtaaactca tatccggtac tgtcgatcaa agcgatcagt catttttaga cgacgatcaa
      421 attgaagctg gatacgttct cacctgcgtt gcttacccaa cttccgattg cgtcatcgaa
      481 acacacaaag aagaagagct ttactaagaa ttctgataat aatctagacc aggcatcaaa
      541 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
      601 acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttataggt
      661 acccgtggat cctctagagt cgacctgcag gcatgcaagc tttttacggc tagctcagtc
      721 ctaggtatag tgctagccca gccagagaaa cactctttaa cagggggtag tatgatgtct
      781 cgtttagata aaagtaaagt gattaacagc gcattagagc tgcttaatga ggtcggaatc
      841 gaaggtttaa caacccgtaa actcgcccag aagctaggtg tagagcagcc tacattgtat
      901 tggcatgtaa aaaataagcg ggctttgctc gacgccttag ccattgagat gttagatagg
      961 caccatactc acttttgccc tttagaaggg gaaagctggc aagatttttt acgtaataac
     1021 gctaaaagtt ttagatgtgc tttactaagt catcgcgatg gagcaaaagt acatttaggt
     1081 acacggccta cagaaaaaca gtatgaaact ctcgaaaatc aattagcctt tttatgccaa
     1141 caaggttttt cactagagaa tgcattatat gcactcagcg cagtggggca ttttacttta
     1201 ggttgcgtat tggaagatca agagcatcaa gtcgctaaag aagaaaggga aacacctact
     1261 actgatagta tgccgccatt attacgacaa gctatcgaat tatttgatca ccaaggtgca
     1321 gagccagcct tcttattcgg ccttgaattg atcatatgcg gattagaaaa acaacttaaa
     1381 tgtgaaagtg ggtcttaagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     1441 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgccct
     1501 agacctaggg cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     1561 tatccacaga atcaggggat aacgcatgca agaacatgtg agcaaaaggc cagcaaaagg
     1621 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1681 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1741 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1801 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1861 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1921 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1981 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2041 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     2101 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2161 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2221 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2281 agtggaacga aaactcacgt taagggattt tggtcatgac tagtgcttgg attctcacca
     2341 ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
     2401 attactggat ctatcaacag gagtccaagc gagctcgata tcaaattacg ccccgccctg
     2461 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
     2521 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
     2581 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     2641 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt
     2701 agggaaatag gccaggtttt caccataaca cgccacatct tgcgaatata tgtgtagaaa
     2761 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     2821 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     2881 catacgaaat tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     2941 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     3001 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
     3061 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     3121 tcctgaaaat ctcgataact caaaaaatac gaccggtagt gatcttattt cattatggtg
     3181 aaagttggaa cctcttacgg gccggccaac
//
