LOCUS       V005247                 8419 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005247
VERSION     V005247
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8419)
  AUTHORS   Rodriguez SA, Yu JJ, Davis G, Arulanandam BP, Klose KE.
  TITLE     Targeted inactivation of francisella tularensis genes by group II
            introns
  JOURNAL   Appl. Environ. Microbiol. 74 (9), 2619-2626 (2008)
   PUBMED   18310413
REFERENCE   2  (bases 1 to 8419)
  AUTHORS   Rodriguez SA, Petrosino JF, Klose KE.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-FEB-2008) Biology, South Texas Center for Emerging
            Infectious Diseases, The University of Texas at San Antonio, One
            UTSA Circle, San Antonio, TX 78249, USA
REFERENCE   3  (bases 1 to 8419)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8419)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Environ. Microbiol."; date: "2008"; volume: "74"; issue: "9"; pages:
            "2619-2626"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (18-FEB-2008) Biology, South Texas Center for Emerging Infectious
            Diseases, The University of Texas at San Antonio, One UTSA Circle,
            San Antonio, TX 78249, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8419
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        142..172
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..195
                     /label="lac operator"
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             216..389
                     /label="lacZ-alpha"
                     /note="LacZ-alpha fragment of beta-galactosidase"
     intron          770..1372
                     /note="Lactococcus lactis LtrB group II intron"
     RBS             1405..1427
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1619..3415
                     /gene="ltrA"
                     /label="Group II intron-encoded protein LtrA"
                     /note="Group II intron-encoded protein LtrA from
                     Lactococcus lactis subsp. cremoris (strain MG1363).
                     Accession#: P0A3U1"
     terminator      3610..3696
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3788..3815
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     rep_origin      4036..4581
                     /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."
     misc_feature    complement(4912..6807)
                     /note="corresponds to region within Francisella tularensis
                     subsp. novicida plasmid, pFNL10, which encodes origin of
                     replication"
     CDS             complement(5273..6292)
                     /codon_start=1
                     /gene="repA"
                     /product="temperature-sensitive RepA"
                     /label="repA"
                     /note="contains amino acid mutation, methionine to
                     isoleucine M120I"
                     /protein_id="ACA63832.1"
                     /translation="MKEVKNVNAGKEIAMSNTLVAGKYSLTKEEQNLIFLVASMVKRED
                     KEFHRYKISLSDLEKATGVKHNRVRLKQLMHSIMSKPVWLNKEQTKIANWFAYIEADPK
                     SSALICEFHWSLMPHIIQLQEYFTKAERQLLFSFKSKYSSRLYLLLKSKLGEQAGYANI
                     VDCVLYVDDMINDFDLPKSYSNRYSNFKNKFLLPALEEINQLSDIFVTYDDKDHHRKHG
                     RKITNIKFTVSKVAETEEEFKQKLLDTKNKEDYIPIEASERLREVVLSDELDLDLYYVR
                     SIFRHHHLHDIERVCDDTWRNWDNKKIKDRKRVFIANIKKLEKKPEENHKLPFGFEEV"
     gene            complement(5273..6292)
                     /gene="repA"
                     /label="repA"
     regulatory      7163..7322
                     /note="region encodes promoter of FTN1451 of Francisella
                     tularensis subsp. novicida"
                     /regulatory_class="promoter"
     CDS             7326..8138
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     regulatory      8181..8419
                     /note="region encodes groELp promoter of Francisella
                     tularensis subsp. holarctica strain, LVS"
                     /regulatory_class="promoter"
ORIGIN
        1 ctcgagaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacattttc acacaggaaa cagctatgac catgattacg gattcactgg
      241 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
      301 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
      361 cccaacagtt gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg gcaccagaag
      421 cggtgccgga aagctggctg gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct
      481 caaactggca gatgcacggt tacgatgcgc ccatctacac caacgtgacc tatcccatta
      541 cggtcaatcc gccgtttgtt cccacggaga atccgacggg ttgttactcg ctcacattta
      601 atgttgatga aagctggcta caggaaggcc agacgcgaat tatttttgat ggcgttaact
      661 cggcgtttca tctgtggtgc aacgggcgct gggtcggtta cggccaggac agtcgtttgc
      721 cgtctgaatt tgacctgagc gcatttttac gcgccggaga aaatgtacaa tctgtaggag
      781 aacctatggg aacgaaacga atgcgatgcc gagaatctga atttaccaag acttaacact
      841 aactggggat accctaaaca agaatgccta atagaaagga ggaaaaaggc tatagcacta
      901 gagcttgaaa atcttgcaag ggtacggagt actcgtagta gtctgagaag ggtaacgccc
      961 tttacatggc aaaggggtac agttattgtg tactaaaatt aaaaattgat tagggaggaa
     1021 aacctcaaaa tgaaaccaac aatggcaatt ttagaaagaa tcagtaaaaa ttcacaagaa
     1081 aatatagacg aagtttttac aagactttat cgttatcttt tacgtccaga tatttattac
     1141 gtgtttttct cgttgggaaa tggcaatgat agcgaaacaa cgtaaaactc ttgttgtatg
     1201 ctttcattgt catcgtcacg tgattcataa acacaagtga atgtcgacag tgaattttta
     1261 cgaacgaaca ataacagagc cgtatactcc gagaggggta cgtacggttc ccgaagaggg
     1321 tggtgcaaac cagtcacagt aatgtgaaca aggcggtacc tccctacttc accatatcat
     1381 tttctgcagc cccctagaaa taattttgtt taactttaag aaggagatat acatatatgg
     1441 ctagatcgtc cattccgaca gcatcgccag tcactatggc gtgctgctag cgctatatgc
     1501 gttgatgcaa tttctatgca ctcgtagtag tctgagaagg gtaacgccct ttacatggca
     1561 aaggggtaca gttattgtgt actaaaatta aaaattgatt agggaggaaa acctcaaaat
     1621 gaaaccaaca atggcaattt tagaaagaat cagtaaaaat tcacaagaaa atatagacga
     1681 agtttttaca agactttatc gttatctttt acgtccagat atttattacg tggcgtatca
     1741 aaatttatat tccaataaag gagcttccac aaaaggaata ttagatgata cagcggatgg
     1801 ctttagtgaa gaaaaaataa aaaagattat tcaatcttta aaagacggaa cttactatcc
     1861 tcaacctgta cgaagaatgt atattgcaaa aaagaattct aaaaagatga gacctttagg
     1921 aattccaact ttcacagata aattgatcca agaagctgtg agaataattc ttgaatctat
     1981 ctatgaaccg gtattcgaag atgtgtctca cggttttaga cctcaacgaa gctgtcacac
     2041 agctttgaaa acaatcaaaa gagagtttgg cggcgcaaga tggtttgtgg agggagatat
     2101 aaaaggctgc ttcgataata tagaccacgt tacactcatt ggactcatca atcttaaaat
     2161 caaagatatg aaaatgagcc aattgattta taaatttcta aaagcaggtt atctggaaaa
     2221 ctggcagtat cacaaaactt acagcggaac acctcaaggt ggaattctat ctcctctttt
     2281 ggccaacatc tatcttcatg aattggataa gtttgtttta caactcaaaa tgaagtttga
     2341 ccgagaaagt ccagaaagaa taacacctga atatcgggag ctccacaatg agataaaaag
     2401 aatttctcac cgtctcaaga agttggaggg tgaagaaaaa gctaaagttc ttttagaata
     2461 tcaagaaaaa cgtaaaagat tacccacact cccctgtacc tcacagacaa ataaagtatt
     2521 gaaatacgtc cggtatgcgg acgacttcat tatctctgtt aaaggaagca aagaggactg
     2581 tcaatggata aaagaacaat taaaactttt tattcataac aagctaaaaa tggaattgag
     2641 tgaagaaaaa acactcatca cacatagcag tcaacccgct cgttttctgg gatatgatat
     2701 acgagtaagg agatctggaa cgataaaacg atctggtaaa gtcaaaaaga gaacactcaa
     2761 tgggagtgta gaactcctta ttcctcttca agacaaaatt cgtcaattta tttttgacaa
     2821 gaaaatagct atccaaaaga aagatagctc atggtttcca gttcacagga aatatcttat
     2881 tcgttcaaca gacttagaaa tcatcacaat ttataattct gaactccgcg ggatttgtaa
     2941 ttactacggt ctagcaagta attttaacca gctcaattat tttgcttatc ttatggaata
     3001 cagctgtcta aaaacgatag cctccaaaca taagggaaca ctttcaaaaa ccatttccat
     3061 gtttaaagat ggaagtggtt cgtgggggat cccgtatgag ataaagcaag gtaagcagcg
     3121 ccgttatttt gcaaatttta gtgaatgtaa atccccttat caatttacgg atgagataag
     3181 tcaagctcct gtattgtatg gctatgcccg gaatactctt gaaaacaggt taaaagctaa
     3241 atgttgtgaa ttatgtggga cgtctgatga aaatacttcc tatgaaattc accatgtcaa
     3301 taaggtcaaa aatcttaaag gcaaagaaaa atgggaaatg gcaatgatag cgaaacaacg
     3361 taaaactctt gttgtatgct ttcattgtca tcgtcacgtg attcataaac acaagtgaga
     3421 tatctcgatc gagcacccgt tctcggagca ctgtccgacc gctttggccg ccgcccagtc
     3481 ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac cacacccgtc
     3541 ctgtggatcg ccaagccgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact
     3601 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     3661 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     3721 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
     3781 attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttcctgtc
     3841 gtcatatcta caagccatcc ccccacagat acggtaaact agcctcgttt ttgcatcagg
     3901 aaagcagaac gccatgagcg gcctcatttc ttattctgag ttacaacagt ccgcaccgct
     3961 gccggtagct ccttccggtg ggcgcggggc atactagttc tagaaatatt ttatctgatt
     4021 aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct ctgaaaacga
     4081 aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga gctaccaact ctttgaaccg
     4141 aggtaactgg cttggaggag cgcagtcacc aaaacttgtc ctttcagttt agccttaacc
     4201 ggcgcatgac ttcaagacta actcctctaa atcaattacc agtggctgct gccagtggtg
     4261 cttttgcatg tctttccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4321 cggactgaac ggggggttcg tgcatacagt ccagcttgga gcgaactgcc tacccggaac
     4381 tgagtgtcag gcgtggaatg agacaaacgc ggccataaca gcggaatgac accggtaaac
     4441 cgaaaggcag gaacaggaga gcgcacgagg gagccgccag ggggaaacgc ctggtatctt
     4501 tatagtcctg tcgggtttcg ccaccactga tttgagcgtc agatttcgtg atgcttgtca
     4561 ggggggcgga gcctatggaa aaacggcttt gccgcggccc tctcacttcc ctgttaagta
     4621 tcttcctggc atcttccagg aaatctccgc cccgttcgta agccatttcc gctcgccgca
     4681 gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc ctgtatcaca
     4741 tattctgctg acgcaccggt gcagcctttt ttctcctgcc acatgaagca cttcactgac
     4801 accctcatca gtgccaacat agtaagccag tatacactcc gctagcgctg atgtccggcg
     4861 gtgcttttgc cgttacgcac caccccgtca gtagctgaac aggagggaca gctagaaatg
     4921 taccaaattt tacctatttt aggatacttt tcatgaagct cttctatttt tccccagtcc
     4981 tttaatagtc tacctttaga gtctcgtaaa tagttatctt tgtgacaggg gcctctttta
     5041 tcttttttaa tgtaactata tgttattcca acgtcactat tactattatc caaatctttt
     5101 ttagcatgcc agtaagaact ttcataactt aactctatct ttcgacctct ttgatataca
     5161 acaataaagc tatagccagt agtaacaacc tgttttactt ttgttaaatc tattaacttc
     5221 ttatttattt ttttatgttt ttttgaaaat ttaaatattt ctatattcat tcctacactt
     5281 cctcaaatcc aaatggtagc ttatgattct cttctggttt cttttctaat ttttttatat
     5341 ttgcaataaa aactcttttt ctatctttga tttttttatt gtcccaattc ctccaagtat
     5401 catcacaaac cctttcaata tcatgtaaat gatgatgtct aaatattgat ctgacataat
     5461 acagatctag gtctagttca tcacttaaca caacttctct aagtctttca gatgcttcga
     5521 ttggtatgta atcctcttta tttttagtat ctaaaagctt ttgcttaaat tcttcttctg
     5581 tctctgctac cttactaact gtaaacttga tatttgtaat cttacgacca tgttttctgt
     5641 gatgatcctt gtcatcatag gttacaaaaa tatccgataa ttgattaatc tcttctagtg
     5701 ctggtaatag gaacttattt ttaaaatttg aatatctgtt gctgtaactt ttaggtaaat
     5761 caaaatcatt aatcatatca tcgacataca atacgcaatc aactatatta gcataccctg
     5821 cttgttcgcc taatttgctt ttgagaagta agtataatct gcttgaatac ttacttttaa
     5881 atgaaaatag taactgtctt tctgctttag taaagtactc ttgtagttgt attatgtgtg
     5941 gcattaatga ccaatgaaac tcgcaaatta aagcactgct tttagggtct gcttcaatat
     6001 atgcaaacca gttagctatc ttcgtttgtt ctttattcag ccatactggc ttagacatta
     6061 ttgagtgcat taattgcttc aatctcactc tgttatgctt aacccctgta gctttttcaa
     6121 gatcagatag gcttatctta tacctgtgaa actctttatc ttctctttta accattgagg
     6181 caactaagaa tattaagttt tgttcttctt ttgtaaggct atactttcct gcaacaagag
     6241 tattagacat agctatttct ttgccagcat ttacattttt aacttctttc atagaactag
     6301 agtcattatc tcgatataca aattctataa aacttctatt agtaaaacaa ctacttcata
     6361 aaaaaaagta gttttaacga tacaaaaagt agttttaaat tcaaaaagtg atacaaaaag
     6421 tagttttaaa ttcaaaaagt gatacaaaaa gtagttttaa attttttaaa aaagtgcttc
     6481 aaagccttat gtagcaatac ttacagagga ttaaaaaaaa atctgacaat atataaagag
     6541 aatatataaa gagaatatct taggggattt taaaaaaatc ccacagactc aaagactttt
     6601 ttgacttttt aaatcctaga aactatactt taagtactta tttaagtaca tggatttaga
     6661 ttatgcaaac cgttaattat tcaactttta gaaatgaact atctgattca atggatagag
     6721 taacaaaaaa tcatagtcct atgattgtaa ctagaggttc aaaaaaagaa gcagttgtta
     6781 tgatgtcgtt agaggatttt aaagctttaa tgcggtattt tatcacagtt aaattgctaa
     6841 cgcagtcagg caccgtgtat gaaatctaac aatgcgctca tcgtcatcct cggcctcgac
     6901 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     6961 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     7021 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     7081 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     7141 ccggcgtaga ggatcgagat cttttgggtt gtcactcatc gtatttggtt tataatttta
     7201 agctaataac ctaattataa ctaattaata gttttgtatc ttgaaaaaat agctataaaa
     7261 cttatttaaa taacgaagat ttttgtgtat aaaatattta taacaaaaaa aaggagacta
     7321 aacatatgag ccatattcaa cgggaaacgt cttgctccag gccgcgatta aattccaaca
     7381 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     7441 caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     7501 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     7561 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     7621 ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     7681 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     7741 gtccttttaa cagcgatcgc gtatttcgtc tcgctcatgc gcaatcacga atgaataacg
     7801 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     7861 ggaaagaaat gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt
     7921 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     7981 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     8041 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     8101 ataaattgca gtttcatttg atgctcgatg agtttttcta aggatccggc tgctaacaaa
     8161 gcccgaaagg aagcggccgc tgaaaaataa acttaattat tatatatgtt atttagctag
     8221 tttttttaat taaagttaaa atcgagagct tgtttgacaa aaaaacaaaa aaatttcttg
     8281 aaaatttttt ttttgactca atatctagac ttgcaagagc ttggaacttt gagattgttc
     8341 taagatgcat acaaattcaa aatgcttaaa caaaaataat ttaacaaagg agtaagattg
     8401 ttatgaacat tcgtccatt
//
