LOCUS       sequence-383719-        3030 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Suicide vector to generate markerless gene deletions in 
            Fusobacterium nucleatum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3030)
  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 3030)
  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-04
FEATURES             Location/Qualifiers
     source          1..3030
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             744..752
                     /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)"
     terminator      904..938
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1010..1636)
                     /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     1541..1560
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    1743..1761
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     rep_origin      2415..3003
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2904..2923
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttcctggcct tttgctggcg aattcttatg gattttaggg taacaaaaaa caccgtattt
       61 ctacgatgtt tttgcttaaa tacttgtttt tagttacaga caaacctgaa gttaactatt
      121 tatcaattcc tgcaattcgt ttacaaaacg gcaaatgtga aatccgtcac atactgcgtg
      181 atgaacttga attgccaaag gaagtataat tttgttatct tctttataat atttccccat
      241 agtaaaaata ggaatcaaat aatcatatcc tttctgcaaa ttcagattaa agccatcgaa
      301 ggttgaccac ggtatcatag atacattaaa aatgttttcc ggagcatttg gctttccttc
      361 cattctatga ttgtttccat accgttgcgt atcactttca taatctgcta aaaatgattt
      421 aaagtcagac ttacactcag tccaaaggct ggaaaatgtt tcagtatcat tgtgaaatat
      481 tgtatagctt ggtatcatct catcatatat ccccaattca ccatcttgat tgattgccgt
      541 cctaaactct gaatggcggt ttacaatcat tgcaatataa taaagcattg caggatatag
      601 tttcattccc ttttccttta tttgtgtgat atccacttta acggtcatgc tgtaggtaca
      661 aggtacactt gcaaagtagt ggtcaaaata ctcttttctg ttccaactat ttttatcaat
      721 tttttcaaat accatttaaa attacctcct taaaatatac tgctttaatt gtataatttt
      781 agctaattat tgtcaagtat ttttaaagtt atgtaaattt tttacttata ttttatcgat
      841 cgagtgctag ccaatcattt tgcggccgct ccagtaatga cctcagaact ccatctggat
      901 ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagcactagt
      961 cataaaaata agaagcctgc atttgcaggc ttcttatttt tatggtacct taagacccac
     1021 tttcacattt aagttgtttt tctaatccgc atatgatcaa ttcaaggccg aataagaagg
     1081 ctggctctgc accttggtga tcaaataatt cgatagcttg tcgtaataat ggcggcatac
     1141 tatcagtagt aggtgtttcc ctttcttctt tagcgacttg atgctcttga tcttccaata
     1201 cgcaacctaa agtaaaatgc cccacagcgc tgagtgcata taatgcattc tctagtgaaa
     1261 aaccttgttg gcataaaaag gctaattgat tttcgagagt ttcatactgt ttttctgtag
     1321 gccgtgtacc taaatgtact tttgctccat cgcgatgact tagtaaagca catctaaaac
     1381 ttttagcgtt attacgtaaa aaatcttgcc agctttcccc ttctaaaggg caaaagtgag
     1441 tatggtgcct atctaacatc tcaatggcta aggcgtcgag caaagcccgc ttatttttta
     1501 catgccaata caatgtaggc tgctctacac ctagcttctg ggcgagttta cgggttgtta
     1561 aaccttcgat tccgacctca ttaagcagct ctaatgcgct gttaatcact ttacttttat
     1621 ctaatctaga catcattaat tcctcctttt tgttgacatt atatcattga tagagttatt
     1681 tgtcaaacta gttttttatt tcgatgccct ggacttcatg aaaaactaaa aaaaatattg
     1741 acactctatc attgatagag tataattaaa ataagcttga tcgtagcgtt aacagatctg
     1801 agctcctctc gaggaaggga agattgccca aggtgagtac taattgtgaa ataaagcgtg
     1861 gtgatttata ttacgctgat ttaagtccag ttgttggttc agagcaggga ggcgttagac
     1921 ctgttctgat aatacaaaat gatgtaggta ataaatatag tcctacagtt atagtatcag
     1981 ctataacatc tcagattaat aaagccaaat tacctactca tgtagagata agttccaatg
     2041 aatatgggct taataaagac tcagtaattc ttttagagca aattagaact atcgataaaa
     2101 agagactgag ggaaaaaatt ggttgtctag atgaaaatat gatgataaaa gtagatgatg
     2161 gacttcaaat aagtttagga ttgtttactt tttaaggatc ctataagttt taataaaact
     2221 ttaaatagaa aaaggcttct ctcatgagaa gtctttttta tttaaaataa atataaaata
     2281 aaatagaggc tataaatagc ctctatttta tgtgagaaat ccctaaataa aaaactagtc
     2341 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2401 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2461 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2521 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     2581 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2641 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2701 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2761 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2821 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2881 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2941 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3001 aaaaacgcca gcaacgcggc ctttttacgg
//
