LOCUS       sequence-406656-        4561 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Backbone for inducible expression system in Fusobacterium nucleatum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4561)
  AUTHORS   Ponath F, Zhu Y, Cosi V, Vogel J
  TITLE     Expanding the genetic toolkit helps dissect a global stress response
            in the early-branching species Fusobacterium nucleatum.
  JOURNAL   Proc Natl Acad Sci U S A. 2022 Oct 4;119(40):e2201460119. doi: 
            10.1073/pnas.2201460119. Epub 2022 Sep 26.
  PUBMED    36161895
REFERENCE   2  (bases 1 to 4561)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2022-10-4"; volume: "119(40):e2201460119. 
            doi"; pages: " 10.1073/pnas.2201460119. Epub 2022 Sep 26"
COMMENT     Sequence Label: pVoPo-03
FEATURES             Location/Qualifiers
     source          1..4561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     55..74
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    257..275
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     rep_origin      506..1094
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     995..1014
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     RBS             1865..1873
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             complement(3389..3400)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     terminator      3979..4013
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(join(4085..4561,1..150))
                     /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"
ORIGIN
        1 ccgcttattt tttacatgcc aatacaatgt aggctgctct acacctagct tctgggcgag
       61 tttacgggtt gttaaacctt cgattccgac ctcattaagc agctctaatg cgctgttaat
      121 cactttactt ttatctaatc tagacatcat taattcctcc tttttgttga cattatatca
      181 ttgatagagt tatttgtcaa actagttttt tatttcgatg ccctggactt catgaaaaac
      241 taaaaaaaat attgacactc tatcattgat agagtataat taaaatggat cctataagtt
      301 ttaataaaac tttaaataga aaaaggcttc tctcatgaga agtctttttt atttaaaata
      361 aatataaaat aaaatagagg ctataaatag cctctatttt atgtgagaaa tccctaaata
      421 aaaaactagt catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
      481 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
      541 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
      601 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
      661 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
      721 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
      781 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
      841 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
      901 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
      961 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     1021 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     1081 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     1141 gaattcttat ggattttagg gtaacaaaaa acaccgtatt tctacgatgt ttttgcttaa
     1201 atacttgttt ttagttacag acaaacctga agttaactat ttatcaattc ctgcaattcg
     1261 tttacaaaac ggcaaatgtg aaatccgtca catactgcgt gatgaacttg aattgccaaa
     1321 ggaagtataa ttttgttatc ttctttataa tatttcccca tagtaaaaat aggaatcaaa
     1381 taatcatatc ctttctgcaa attcagatta aagccatcga aggttgacca cggtatcata
     1441 gatacattaa aaatgttttc cggagcattt ggctttcctt ccattctatg attgtttcca
     1501 taccgttgcg tatcactttc ataatctgct aaaaatgatt taaagtcaga cttacactca
     1561 gtccaaaggc tggaaaatgt ttcagtatca ttgtgaaata ttgtatagct tggtatcatc
     1621 tcatcatata tccccaattc accatcttga ttgattgccg tcctaaactc tgaatggcgg
     1681 tttacaatca ttgcaatata ataaagcatt gcaggatata gtttcattcc cttttccttt
     1741 atttgtgtga tatccacttt aacggtcatg ctgtaggtac aaggtacact tgcaaagtag
     1801 tggtcaaaat actcttttct gttccaacta tttttatcaa ttttttcaaa taccatttaa
     1861 aattacctcc ttaaaatata ctgctttaat tgtataattt tagctaatta ttgtcaagta
     1921 tttttaaagt tatgtaaatt ttttacttat attttatcga tcgttctaga tattcttttt
     1981 taaacttttc aaatgcttca aattctacat ctgcaacttc tacatctttt ccagtatttc
     2041 caaacttcac acaatttttg taaacatttc ttaatttttc caagtattct atgtcttcct
     2101 tttcttcttt aacatactct tgtaactgtg tcatcaaatt agttttgtct tccacattcc
     2161 ccctttttct tgtaattgcc ttatttaatt cttcaaatct tattgttttc attttattgt
     2221 acctccaatt tttttatata tctatgttaa aaaacatatt gacaattgat taaaacaatg
     2281 atattatggt cggttgagct cattgagctg agtgcagact ttcccccttg taaaaagggg
     2341 aaaaaacgat tgataatgat acaatggtcg tatatcgcta tgttcgcatc gtttaactta
     2401 ctttatcgca tatttaatta tatgccagtg ttagccctgc ccaacaccta gagttttagc
     2461 ttactttcta aattataacc ctaataaagt acccactcag agcacctcca agacttttaa
     2521 aggtagcctc gcaccttgct acaattcata ataccaccat tacggtgcta taccgtttcg
     2581 agcagaacgg ctatacatac gaatttatct tatgtataaa atttgttaca aatataatag
     2641 taagtttttt ttatataaaa aagcagatcg cttttatttt ttttgcaacc tgcttcacaa
     2701 ctcatttttt tactttagta aataaatcct ttacatcttt taaaaaatat ggtatattaa
     2761 gtcaaagcag gcataacata tatgttatgt tttatagtaa aaatggacta ttcaactatt
     2821 taaaaaatgg tagaatacaa ataattttcc tggcagtcaa ttatttttta aaaattttta
     2881 ctattatatt tttaaatttt aaattttaga tagagttatt tggcgataac tctttttgtc
     2941 ttacttctga gtttagcata ttttttttgt tttgtaaatc catttcttta tttttcttta
     3001 caacataaac tttttttact tctctataaa tcttctcacc agttgaaaca tcaatttcta
     3061 acagttctct agcaataaaa tttttaatat agagtaaatc tttttcatct gtaagtatat
     3121 cattcatttc atcaaaatcc aacttttttt tggtttcttt tggttttatt aaatttcgtg
     3181 aacagaaata ggcttttttt tgatttgctt tcttttctaa gtctttcaca aaatacttcg
     3241 ttatatacac ccccacagct tcatttttaa cagtttcttt aatcttattt atttttataa
     3301 atccattcgc ccacaattcc ccaagcttct tattatctat atatgggacc gagaacagca
     3361 ccaaatgata atggattgct cccctctgct ggagttccca agttgcaata tatttaagct
     3421 gttgattttt caagtacctt ttcaatctct taataaataa agtaaattca cgatttgccc
     3481 tttctatatc ctgtatattt tctttaaaag taagagttaa aaaggtactt ttatcatcat
     3541 aattacaatc aattatcctt tttatttcaa atcgcttgtc ctggtaatat gctttccttc
     3601 gttgcatact ttcagctttt tcaatgctac ttaattcttc aaaatctctt ctttttcttc
     3661 ctggtcttct tttaaagtca aaagcagtta aagcatcaat attgaaaatg tattcagtta
     3721 tattttttga ttttatttct ttagtgttat aaaatttcaa ctcaataaat tcccccttta
     3781 ttaatctgca taatcttttt tataatgtcc tttacaacct aatattctta atctatcatt
     3841 ttttaagtca taaattagtc tatgttcatc acttattctt ctgctataaa tagccctatt
     3901 tttaaaatgt tttaatctct caggcttacc agtaccgcgg ccgctccagt aatgacctca
     3961 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
     4021 aatccaagca ctagtcataa aaataagaag cctgcatttg caggcttctt atttttatgg
     4081 taccttaaga cccactttca catttaagtt gtttttctaa tccgcatatg atcaattcaa
     4141 ggccgaataa gaaggctggc tctgcacctt ggtgatcaaa taattcgata gcttgtcgta
     4201 ataatggcgg catactatca gtagtaggtg tttccctttc ttctttagcg acttgatgct
     4261 cttgatcttc caatacgcaa cctaaagtaa aatgccccac agcgctgagt gcatataatg
     4321 cattctctag tgaaaaacct tgttggcata aaaaggctaa ttgattttcg agagtttcat
     4381 actgtttttc tgtaggccgt gtacctaaat gtacttttgc tccatcgcga tgacttagta
     4441 aagcacatct aaaactttta gcgttattac gtaaaaaatc ttgccagctt tccccttcta
     4501 aagggcaaaa gtgagtatgg tgcctatcta acatctcaat ggctaaggcg tcgagcaaag
     4561 c
//
