LOCUS       sequence-232431-        7800 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 7800)
  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 7800)
  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: pFNMB2-msfGFP
FEATURES             Location/Qualifiers
     source          1..7800
                     /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    1924..1942
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      2837..2923
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3015..3042
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(3644..4015)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            4048..4157
                     /note="incP origin of transfer"
     CDS             5100..5900
                     /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(5160..5179)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5770..5789
                     /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 ttttgactca atatctagac
     1921 ttgtccctat cagtgataga gactgcagat ttaagaagga gatacgcgta tgggatctaa
     1981 aggtgaagaa ctgttcaccg gtgttgttcc gatcctggtt gaactggatg gtgatgttaa
     2041 cggccacaaa ttctctgttc gtggtgaagg tgaaggtgat gcaaccaacg gtaaactgac
     2101 cctgaaattc atctgcacta ccggtaaact gccggttcca tggccgactc tggtgactac
     2161 cctgacctat ggtgttcagt gtttttctcg ttacccggat cacatgaagc agcatgattt
     2221 cttcaaatct gcaatgccgg aaggttatgt acaggagcgc accatttctt tcaaagacga
     2281 tggcacctac aaaacccgtg cagaggttaa atttgaaggt gatactctgg tgaaccgtat
     2341 tgaactgaaa ggcattgatt tcaaagagga cggcaacatc ctgggccaca aactggaata
     2401 taacttcaac tcccataacg tttacatcac cgcagacaaa cagaagaacg gtatcaaagc
     2461 taacttcaaa attcgccata acgttgaaga cggtagcgta cagctggcgg accactacca
     2521 gcagaacact ccgatcggtg atggtccggt tctgctgccg gataaccact acctgtccac
     2581 ccagtctaag ctgtccaaag acccgaacga aaagcgcgac cacatggtgc tgctggagtt
     2641 cgttactgca gcaggtatca cgcacggcat ggatgagctc tacaaataac ccggggcggt
     2701 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     2761 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     2821 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     2881 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     2941 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     3001 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt ttgaattctt agatccagcc
     3061 gaccaggctt tccacgcccg cgtgccgctc catgtcgttc gcgcggttct cggaaacgcg
     3121 ctgccgcgtt tcgtgattgt cacgctcaag cccgtagtcc cgttcgagcg tcgcgcagag
     3181 gtcagcgagg gcgcggtagg cccgatacgg ctcatggatg gtgtttcggg tcgggtgaat
     3241 cttgttgatg gcgatatgga tgtgcaggtt gtcggtgtcg tgatgcacgg cactgacgcg
     3301 ctgatgctcg gcgaagccaa gcccagcgca gatgcggtcc tcaatcgcgc gcaacgtctc
     3361 cgcgtcgggc ttctctcccg cgcggaagct aaccagcagg tgataggtct tgtcggcctc
     3421 ggaacgggtg ttgccgtgct gggtcgccat cacctcggcc atgacagcgg gcagggtgtt
     3481 tgcctcgcag ttcgtgacgc gcacgtgacc caggcgctcg gtcttgcctt gctcgtcggt
     3541 gatgtacttc accagctccg cgaagtcgct cttcttgatg gagcgcatgg ggacgtgctt
     3601 ggcaatcacg cgcacccccc ggccgtttta gcggctaaaa aagtcatggc tctgccctcg
     3661 ggcggaccac gcccatcatg accttgccaa gctcgtcctg cttctcttcg atcttcgcca
     3721 gcagggcgag gatcgtggca tcaccgaacc gcgccgtgcg cgggtcgtcg gtgagccaga
     3781 gtttcagcag gccgcccagg cggcccaggt cgccattgat gcgggccagc tcgcggacgt
     3841 gctcatagtc cacgacgccc gtgattttgt agccctggcc gacggccagc aggtaggccg
     3901 acaggctcat gccggccgcc gccgcctttt cctcaatcgc tcttcgttcg tctggaaggc
     3961 agtacacctt gataggtggg ctgcccttcc tggttggctt ggtttcatca gccatccgct
     4021 tgccctcatc tgttacgccg gcggtagccg gccagcctcg cagagcagga ttcccgttga
     4081 gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg ggtgggccta
     4141 cttcacctat cctgcccggc tgacgccgtt ggatacacca aggaaagtct acacgaaccc
     4201 tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa atcgctataa
     4261 tgaccccgaa gcagggttat gcagcggaaa agcgctgctt ccctgctgtt ttgtggaata
     4321 tctaccgact ggaaacaggc aaatgcagga aattactgaa ctgaggggac aggcgagaga
     4381 cgatgccaaa gagctacacc gacgagctgg ccgagtgggt tgaatcccgc gcggccaaga
     4441 agcgccggcg tgatgaggct gcggttgcgt tcctggcggt gagggcggat gtcgaggcgg
     4501 cgttagcgtc cggctatgcg ctcgtcacca tttgggagca catgcgggaa acggggaagg
     4561 tcaagttctc ctacgagacg ttccgctcgc acgccaggcg gcacatcaag gccaagcccg
     4621 ccgatgtgcc cgcaccgcag gccaaggctg cggaacccgc gccggcaccc aagacgccgg
     4681 agccacggcg gccgaagcag gggggcaagg ctgaaaagcc ggcccccgct gcggccccga
     4741 ccggcttcac cttcaaccca acaccggaca aaaaggattg acctgcaggc caaaaaactc
     4801 aaatcctatt gtatggatta gtcgagctaa aaagctcata ttttttatat tcaaactata
     4861 tacccttatt tacaagcttt gaaaaataaa cttaattatt atatatgtta tttagctagt
     4921 ttttttaatt aaagttaaaa tcgagagctt gtttgacaaa aaaacaaaaa aatttcttga
     4981 aaattttttt tttgactcaa tatctagact tgcaagagct tggagctttg agattgttct
     5041 aagatgcaca caaattcaaa acgcttaaac aaaaaactta atttaacaaa ggagtaagag
     5101 gatccatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5161 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5221 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5281 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5341 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5401 ggcaggatct cctgtcatcc caccttgctc ctgccgagaa agtatccatc atggctgatg
     5461 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5521 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5581 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     5641 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5701 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5761 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5821 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5881 ttcttgacga gttcttctga attgtcgact taaaatcctc taacgacatc ataacaactg
     5941 cttctttttt tgaacctcta gttacaatca taggactatg attttttgtt actctatccg
     6001 ttgaatcaga tagttcattt ctaaaagttg aataattaac ggtttgcata atctaaatcc
     6061 atgtacttaa ataagtactt aaagtatagt ttctaggatt taaaaagtca aaaaagtctt
     6121 tgagtctgtg ggattttttt aaaatcccct aagatattct ctttatatat tctctttata
     6181 tattgtcaga ttttttttta atcctctgta agtattgcta cataaggctt tgaagcactt
     6241 ttttaaaaaa tttaaaacta ctttttgtat cactttttga atttaaaact actttttgta
     6301 tcactttttg aatttaaaac tactttttgt atcgttaaaa ctactttttt ttatgaagta
     6361 gttgttttac taatagaagt tttatagaat ttgtatatcg agataatgac tctagttcta
     6421 tgaaagaagt taaaaatgta aatgctggca aagaaatagc tatgtctaat actcttgttg
     6481 caggaaagta tagccttaca aaagaagaac aaaacttaat attcttagtt gcctcaatgg
     6541 ttaaaagaga ggataaagag tttcacaggt ataagataag cctatctgat cttgaaaaag
     6601 ctacaggggt taagcataac agagtgagat tgaagcaatt aatgcactca ataatgtcta
     6661 agccagtatg gctgaataaa gaacaaacga agatagctaa ctggtttgca tatattgaag
     6721 cagaccctaa aagcagtgct ttaatttgcg agtttcattg gtcattaatg ccacacatga
     6781 tacaactaca agagtacttt actaaagcag aaagacagtt actattttca tttaaaagta
     6841 agtattcaag cagattatac ttacttctca aaagcaaatt aggcgaacaa gcagggtatg
     6901 ctaatatagt tgattgcgta ttgtatgtcg atgatatgat taatgatttt gatttaccta
     6961 aaagttacag caacagatat tcaaatttta aaaataagtt cctattacca gcactagaag
     7021 agattaatca attatcggat ctttttgtaa cctatgatga caaggatcat cacagaaaac
     7081 atggtcgtaa gattacaaat atcaagttta cagttagtaa ggtagcagag acagaagaag
     7141 aatttaagca gaagctttta gatactaaaa ataaagagga ttacatacca atcgaagcat
     7201 ctgaaagact tagagaagtt gtgttaagtg atgaactaga cctagatctg tattatgtca
     7261 gatcaatatt tagacatcat catttacatg atattgaaag ggtttgtgat gatacttgga
     7321 ggaattggga caataaaaaa atcaaagata gaaaaagagt ttttattgca aatataaaaa
     7381 aattagaaaa gaaaccagaa gagaatcata agctaccatt tggatttgag gaagtgtagg
     7441 aatgaatata gaaatattta aattttcaaa aaaacataaa aaaataaata agaagttgat
     7501 agatttaaca aaagtaaaac aggttgttac tactggctat agctttattg ttgtatatca
     7561 aagaggtcga aagatagagt taagttatga aagttcttac tggcatgcta aaaaagattt
     7621 ggataatagt aatagtgacg ttggaataac atatagttac attaaaaaag ataaaagagg
     7681 cccctgtcac aaagataact atttacgaga ctctaaaggt agactattaa aggactgggg
     7741 aaaaatagaa gagcatcatg aaaagtatcc taaaataggt aaaatttggt acatttctag
//
