LOCUS       sequence-220818-        7797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mobilizable plasmid for tetracycline inducible gene expression in 
            Francisella novicida.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7797)
  AUTHORS   Brodmann M, Heilig R, Broz P, Basler M
  TITLE     Mobilizable Plasmids for Tunable Gene Expression in Francisella 
            novicida.
  JOURNAL   Front Cell Infect Microbiol. 2018 Aug 31;8:284. doi: 
            10.3389/fcimb.2018.00284. eCollection 2018.
  PUBMED    30234022
REFERENCE   2  (bases 1 to 7797)
  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: "Front Cell 
            Infect Microbiol."; date: "2018-08-31"; volume: "8:284. doi"; pages:
            " 10.3389/fcimb.2018.00284. eCollection 2018"
COMMENT     Sequence Label: pFNMB1-msfGFP
FEATURES             Location/Qualifiers
     source          1..7797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(197..214)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(331..876)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(411..430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(920..1546)
                     /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     1451..1470
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    1923..1941
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      2834..2920
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3012..3039
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(3641..4012)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            4045..4154
                     /note="incP origin of transfer"
     CDS             5097..5897
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="GSMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5157..5176)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5767..5786
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 ctgtccctcc tgttcagcta ctgacggggt ggtgcgtaac ggcaaaagca ccgccggaca
       61 tcagcgctag cggagtgtat actggcttac tatgttggca ctgatgaggg tgtcagtgaa
      121 gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg tcagcagaat atgtgataca
      181 ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag
      241 cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca
      301 gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca
      361 tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca
      421 ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg
      481 tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt
      541 aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg
      601 ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg
      661 cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc
      721 taaactgtaa ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa
      781 gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca
      841 gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata
      901 aaatatttct agaactagtt taagacccac tttcacattt aagttgtttt tctaatccgc
      961 atatgatcaa ttcaaggccg aataagaagg ctggctctgc accttggtga tcaaataatt
     1021 cgatagcttg tcgtaataat ggcggcatac tatcagtagt aggtgtttcc ctttcttctt
     1081 tagcgacttg atgctcttga tcttccaata cgcaacctaa agtaaaatgc cccacagcgc
     1141 tgagtgcata taatgcattc tctagtgaaa aaccttgttg gcataaaaag gctaattgat
     1201 tttcgagagt ttcatactgt ttttctgtag gccgtgtacc taaatgtact tttgctccat
     1261 cgcgatgact tagtaaagca catctaaaac ttttagcgtt attacgtaaa aaatcttgcc
     1321 agctttcccc ttctaaaggg caaaagtgag tatggtgcct atctaacatc tcaatggcta
     1381 aggcgtcgag caaagcccgc ttatttttta catgccaata caatgtaggc tgctctacac
     1441 ctagcttctg ggcgagttta cgggttgtta aaccttcgat tccgacctca ttaagcagct
     1501 ctaatgcgct gttaatcact ttacttttat ctaatctaga catcattatg gtccatggat
     1561 ccgccattag ttattattct ccgtttttta tcagttttta tatcaaatta atccaaacac
     1621 tcaataagag catttgctat gcaaaaatag cattagacaa taataatatc aaaaaagcaa
     1681 agaaaagcaa agaaaaacta attagttatc agaagcacca aaaactgatt taaaagccac
     1741 atagagatag ttacacatag aataaatagt tagtataacc gcaacataaa gcattaagaa
     1801 acctaagaat ataaaccaag tattgaaact cgtatagtca tagaagcttg catgcctgca
     1861 ggtcgacaaa aaaacaaaaa aatttcttga aaattttttt tttgactcaa tatctagact
     1921 tgtccctatc agtgatagag agtcgaccct cgagaggaga acgcgtatgg gatctaaagg
     1981 tgaagaactg ttcaccggtg ttgttccgat cctggttgaa ctggatggtg atgttaacgg
     2041 ccacaaattc tctgttcgtg gtgaaggtga aggtgatgca accaacggta aactgaccct
     2101 gaaattcatc tgcactaccg gtaaactgcc ggttccatgg ccgactctgg tgactaccct
     2161 gacctatggt gttcagtgtt tttctcgtta cccggatcac atgaagcagc atgatttctt
     2221 caaatctgca atgccggaag gttatgtaca ggagcgcacc atttctttca aagacgatgg
     2281 cacctacaaa acccgtgcag aggttaaatt tgaaggtgat actctggtga accgtattga
     2341 actgaaaggc attgatttca aagaggacgg caacatcctg ggccacaaac tggaatataa
     2401 cttcaactcc cataacgttt acatcaccgc agacaaacag aagaacggta tcaaagctaa
     2461 cttcaaaatt cgccataacg ttgaagacgg tagcgtacag ctggcggacc actaccagca
     2521 gaacactccg atcggtgatg gtccggttct gctgccggat aaccactacc tgtccaccca
     2581 gtctaagctg tccaaagacc cgaacgaaaa gcgcgaccac atggtgctgc tggagttcgt
     2641 tactgcagca ggtatcacgc acggcatgga tgagctctac aaataacccg gggcggtctg
     2701 ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac
     2761 tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg
     2821 aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     2881 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa
     2941 cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca
     3001 tcaaattaag cagaaggcca tcctgacgga tggccttttg aattcttaga tccagccgac
     3061 caggctttcc acgcccgcgt gccgctccat gtcgttcgcg cggttctcgg aaacgcgctg
     3121 ccgcgtttcg tgattgtcac gctcaagccc gtagtcccgt tcgagcgtcg cgcagaggtc
     3181 agcgagggcg cggtaggccc gatacggctc atggatggtg tttcgggtcg ggtgaatctt
     3241 gttgatggcg atatggatgt gcaggttgtc ggtgtcgtga tgcacggcac tgacgcgctg
     3301 atgctcggcg aagccaagcc cagcgcagat gcggtcctca atcgcgcgca acgtctccgc
     3361 gtcgggcttc tctcccgcgc ggaagctaac cagcaggtga taggtcttgt cggcctcgga
     3421 acgggtgttg ccgtgctggg tcgccatcac ctcggccatg acagcgggca gggtgtttgc
     3481 ctcgcagttc gtgacgcgca cgtgacccag gcgctcggtc ttgccttgct cgtcggtgat
     3541 gtacttcacc agctccgcga agtcgctctt cttgatggag cgcatgggga cgtgcttggc
     3601 aatcacgcgc accccccggc cgttttagcg gctaaaaaag tcatggctct gccctcgggc
     3661 ggaccacgcc catcatgacc ttgccaagct cgtcctgctt ctcttcgatc ttcgccagca
     3721 gggcgaggat cgtggcatca ccgaaccgcg ccgtgcgcgg gtcgtcggtg agccagagtt
     3781 tcagcaggcc gcccaggcgg cccaggtcgc cattgatgcg ggccagctcg cggacgtgct
     3841 catagtccac gacgcccgtg attttgtagc cctggccgac ggccagcagg taggccgaca
     3901 ggctcatgcc ggccgccgcc gccttttcct caatcgctct tcgttcgtct ggaaggcagt
     3961 acaccttgat aggtgggctg cccttcctgg ttggcttggt ttcatcagcc atccgcttgc
     4021 cctcatctgt tacgccggcg gtagccggcc agcctcgcag agcaggattc ccgttgagca
     4081 ccgccaggtg cgaataaggg acagtgaaga aggaacaccc gctcgcgggt gggcctactt
     4141 cacctatcct gcccggctga cgccgttgga tacaccaagg aaagtctaca cgaacccttt
     4201 ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc gctataatga
     4261 ccccgaagca gggttatgca gcggaaaagc gctgcttccc tgctgttttg tggaatatct
     4321 accgactgga aacaggcaaa tgcaggaaat tactgaactg aggggacagg cgagagacga
     4381 tgccaaagag ctacaccgac gagctggccg agtgggttga atcccgcgcg gccaagaagc
     4441 gccggcgtga tgaggctgcg gttgcgttcc tggcggtgag ggcggatgtc gaggcggcgt
     4501 tagcgtccgg ctatgcgctc gtcaccattt gggagcacat gcgggaaacg gggaaggtca
     4561 agttctccta cgagacgttc cgctcgcacg ccaggcggca catcaaggcc aagcccgccg
     4621 atgtgcccgc accgcaggcc aaggctgcgg aacccgcgcc ggcacccaag acgccggagc
     4681 cacggcggcc gaagcagggg ggcaaggctg aaaagccggc ccccgctgcg gccccgaccg
     4741 gcttcacctt caacccaaca ccggacaaaa aggattgacc tgcaggccaa aaaactcaaa
     4801 tcctattgta tggattagtc gagctaaaaa gctcatattt tttatattca aactatatac
     4861 ccttatttac aagctttgaa aaataaactt aattattata tatgttattt agctagtttt
     4921 tttaattaaa gttaaaatcg agagcttgtt tgacaaaaaa acaaaaaaat ttcttgaaaa
     4981 tttttttttt gactcaatat ctagacttgc aagagcttgg agctttgaga ttgttctaag
     5041 atgcacacaa attcaaaacg cttaaacaaa aaacttaatt taacaaagga gtaagaggat
     5101 ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5161 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5221 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5281 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5341 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5401 aggatctcct gtcatcccac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5461 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5521 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5581 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5641 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5701 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5761 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5821 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5881 ttgacgagtt cttctgaatt gtcgacttaa aatcctctaa cgacatcata acaactgctt
     5941 ctttttttga acctctagtt acaatcatag gactatgatt ttttgttact ctatccgttg
     6001 aatcagatag ttcatttcta aaagttgaat aattaacggt ttgcataatc taaatccatg
     6061 tacttaaata agtacttaaa gtatagtttc taggatttaa aaagtcaaaa aagtctttga
     6121 gtctgtggga tttttttaaa atcccctaag atattctctt tatatattct ctttatatat
     6181 tgtcagattt ttttttaatc ctctgtaagt attgctacat aaggctttga agcacttttt
     6241 taaaaaattt aaaactactt tttgtatcac tttttgaatt taaaactact ttttgtatca
     6301 ctttttgaat ttaaaactac tttttgtatc gttaaaacta ctttttttta tgaagtagtt
     6361 gttttactaa tagaagtttt atagaatttg tatatcgaga taatgactct agttctatga
     6421 aagaagttaa aaatgtaaat gctggcaaag aaatagctat gtctaatact cttgttgcag
     6481 gaaagtatag ccttacaaaa gaagaacaaa acttaatatt cttagttgcc tcaatggtta
     6541 aaagagagga taaagagttt cacaggtata agataagcct atctgatctt gaaaaagcta
     6601 caggggttaa gcataacaga gtgagattga agcaattaat gcactcaata atgtctaagc
     6661 cagtatggct gaataaagaa caaacgaaga tagctaactg gtttgcatat attgaagcag
     6721 accctaaaag cagtgcttta atttgcgagt ttcattggtc attaatgcca cacatgatac
     6781 aactacaaga gtactttact aaagcagaaa gacagttact attttcattt aaaagtaagt
     6841 attcaagcag attatactta cttctcaaaa gcaaattagg cgaacaagca gggtatgcta
     6901 atatagttga ttgcgtattg tatgtcgatg atatgattaa tgattttgat ttacctaaaa
     6961 gttacagcaa cagatattca aattttaaaa ataagttcct attaccagca ctagaagaga
     7021 ttaatcaatt atcggatctt tttgtaacct atgatgacaa ggatcatcac agaaaacatg
     7081 gtcgtaagat tacaaatatc aagtttacag ttagtaaggt agcagagaca gaagaagaat
     7141 ttaagcagaa gcttttagat actaaaaata aagaggatta cataccaatc gaagcatctg
     7201 aaagacttag agaagttgtg ttaagtgatg aactagacct agatctgtat tatgtcagat
     7261 caatatttag acatcatcat ttacatgata ttgaaagggt ttgtgatgat acttggagga
     7321 attgggacaa taaaaaaatc aaagatagaa aaagagtttt tattgcaaat ataaaaaaat
     7381 tagaaaagaa accagaagag aatcataagc taccatttgg atttgaggaa gtgtaggaat
     7441 gaatatagaa atatttaaat tttcaaaaaa acataaaaaa ataaataaga agttgataga
     7501 tttaacaaaa gtaaaacagg ttgttactac tggctatagc tttattgttg tatatcaaag
     7561 aggtcgaaag atagagttaa gttatgaaag ttcttactgg catgctaaaa aagatttgga
     7621 taatagtaat agtgacgttg gaataacata tagttacatt aaaaaagata aaagaggccc
     7681 ctgtcacaaa gataactatt tacgagactc taaaggtaga ctattaaagg actggggaaa
     7741 aatagaagag catcatgaaa agtatcctaa aataggtaaa atttggtaca tttctag
//
