LOCUS       40924_46543        5969 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWFRT-tel, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5969)
  AUTHORS   Barrett AR, Kang Y, Inamasu KS, Son MS, Vukovich JM, Hoang TT.
  TITLE     Genetic tools for allelic replacement in Burkholderia species
  JOURNAL   Appl. Environ. Microbiol. 74 (14), 4498-4508 (2008)
  PUBMED    18502918
REFERENCE   2  (bases 1 to 5969)
  AUTHORS   Barrett AR, Kang Y, Vukovich JM, Son MS, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-DEC-2007) Microbiology, University of Hawaii at Manoa,
            2538 McCarthy Mall Snyder Hall Rm. 310, Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 5969)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5969)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2008"; volume: "74"; issue: "14"; 
            pages: "4498-4508"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-DEC-2007) Microbiology, University of Hawaii at Manoa, 2538 
            McCarthy Mall Snyder Hall Rm. 310, Honolulu, HI 96822, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /label=AmpR promoter
     CDS             201..1058
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1232..1820
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2144..2174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /label=lac operator
                     /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)."
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2289..2356
                     /label=Flip recognition target
                     /note="Flip recognition target"
     protein_bind    complement(2306..2353)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      2393..2434
                     /regulatory_class="promoter"
     RBS             2445..2467
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2474..3247
                     /codon_start=1
                     /gene="kilA"
                     /product="KilA"
                     /label=kilA
                     /note="confers tellurite resistance"
                     /protein_id="ABY57996.1"
                     /translation="MEEQSVNMARLKGEVLPALFASPATIGEYGAGIDGADSLNELSNL
                     MEHGAVAALADKISQIVAKLADADPRKIAEKPTWFEKMLGREVERQVRYQVARKTLDQL
                     LDEAEGVAQRVRDTLRALDDMLNTHEAEVDRLRAYIQAGREFLDENPEAGAAKAGVIEF
                     DKPRERFARKLANLATLMASHEMSVTQMKLTRAQAVDMLDRFSETASVLVPVWRQHTLA
                     LITTKNMNPAMVAEAAKAHQALMRSLSQSLEGINQ"
     gene            2474..3247
                     /gene="kilA"
                     /label=kilA
     CDS             3265..4401
                     /codon_start=1
                     /gene="telA"
                     /product="TelA"
                     /label=telA
                     /note="confers tellurite resistance"
                     /protein_id="ABY57997.1"
                     /translation="MNALKTTHDAKAPIVAFDMTPATLRELGLQESDVPEVHAVAQRIE
                     VGSPQTVAEFGRDVAEHTSRYADSLLDQVRNSDLDEAGEKLTQVVAKARSLNVGPLSDN
                     RSRLPLIGPLIDRFRVRSTGFMARFDTTREQIEHLVSEVQTTQQGIAQRNASLDEMFAA
                     VREEHRLLGVHIAAGKVRLAELREQAEGLRGNVGNDPGRVQELADLDAMVANLDKRIGD
                     LIALQHSAMQSLPTIRMIQANNQMLVDKFHTIREITVPAWKRQFMLALSLNEQKNAVEL
                     ATAIDDTTNDLMKRNAALLHRTSVETAKENQRLVIDVDTLKQVQTTLIKTVEDVIRIQQ
                     EGVQKRKDAEKQIAAMRGDLQAKLTRQPVRELAQQESV"
     gene            3265..4401
                     /gene="telA"
                     /label=telA
     CDS             4398..5351
                     /codon_start=1
                     /gene="telB"
                     /product="TelB"
                     /label=telB
                     /note="confers tellurite resistance"
                     /protein_id="ABY57998.1"
                     /translation="MNATNTDVFAQVGGLEARGAKMKKRGTRFLIAALAVLAIAGIGAV
                     TGWAISPSATPGSIDVPQVLASTFSDQVPGSEGGGLGGGLPFTSAVGAFTDFMAGPAIF
                     TLGILGIVVAGAVLVFGGEFCGFVRSVCMMVIAVSMIFVSSNLVKGILGGDHDAGPAEP
                     SPRARFMAAVEAKDFARVQELIEARGAKSAADYVLAQLAVAEGLDRKPGARVVVGKAAG
                     SMAMPPAALGFTPRGEAAYAIERSAYGEPRSSIAKQYQQEWNRKAATWWAMAGVAGIIG
                     AILAAAATGFVGLAVSIRNRVKRVRDLLVMEPGAEP"
     gene            4398..5351
                     /gene="telB"
                     /label=telB
     terminator      complement(5369..5400)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
     protein_bind    complement(5461..5508)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(5575..5591)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gcatgcctgc aggtcgactc taggatcccc gggtaccgag
     2281 ctcgaattag cttcaaaagc gctctgaagt tcctatactt tctagagaat aggaacttcg
     2341 gaataggaac ttcaagatcc cctgattccc tttgtcaaca gcaatggata tcagccgttg
     2401 acttagttgg tatttccgga atatcatgct gggttccgaa taattttgtt taactttaag
     2461 aaggagatat accatggaag aacaaagcgt gaacatggcg cgattgaagg gggaggtttt
     2521 gcccgccctc ttcgcgtcgc cggcgacgat tggcgagtac ggggccggca tcgacggggc
     2581 ggattccctc aacgagctgt cgaatctgat ggagcacggc gcagttgccg cgctggccga
     2641 caaaatcagc cagatcgtgg cgaagctggc cgacgcggac ccccgcaaga tcgcggaaaa
     2701 gcctacctgg ttcgagaaga tgcttggccg tgaggttgaa cgccaggtga ggtatcaggt
     2761 cgcccgcaag acgctcgacc agttgctgga cgaagccgag ggcgtagcgc agcgcgtgcg
     2821 ggacacgttg cgcgccttgg atgacatgct caatacgcat gaggccgagg tagaccggct
     2881 cagagcctac attcaagccg ggcgcgagtt cctggacgag aaccccgagg ccggcgcggc
     2941 caaggccggc gtgatcgagt tcgacaagcc gcgcgaacgc ttcgcgcgca agctcgccaa
     3001 cctggcaacc ctcatggcgt cccatgaaat gagcgtcact cagatgaagc tcacgcgggc
     3061 gcaggccgtg gacatgctgg accgcttctc tgaaacggca tccgtcctgg tgcccgtctg
     3121 gcgtcagcac accctcgcgc tcatcaccac caagaacatg aatccggcaa tggtcgccga
     3181 ggcggccaaa gctcaccagg cgctcatgcg gagcctttcg cagagcctgg aaggcatcaa
     3241 ccaataacac ggcgggagaa ccctatgaac gcactgaaaa cgacgcacga cgccaaggcc
     3301 cctatcgtcg ccttcgacat gaccccggca accctgcgcg agctgggctt gcaggaaagc
     3361 gacgtgccgg aagtccatgc ggtcgcgcag cggatcgagg tcggcagtcc gcagaccgtt
     3421 gccgagttcg gccgcgacgt ggccgagcac acgtcccgct acgccgatag cctgctggac
     3481 caggtgcgca acagcgacct ggacgaagca ggcgagaaac tgacccaggt tgtcgccaag
     3541 gcccgttccc tgaacgtcgg ccctttgtcc gacaaccgtt cccgcctgcc cctgattggc
     3601 ccgctgatcg accgcttccg cgtccgttcg acgggcttca tggcgcgctt cgacacgacc
     3661 cgcgagcaga tcgaacacct ggtcagcgaa gtgcagacca cccagcaagg catcgcgcag
     3721 cgcaatgcct cgctcgacga aatgttcgca gccgtgcgcg aggaacaccg ccttcttggc
     3781 gtccacatcg cggccggcaa ggtccgcctt gccgagctgc gcgagcaggc cgagggtctg
     3841 cgcggcaatg tcgggaacga ccctggccgc gtgcaggagc tggccgacct cgatgcgatg
     3901 gttgccaacc tggacaagcg catcggcgac ctgatcgcct tgcaacattc ggccatgcag
     3961 agcctgccga ccatccgcat gatccaggcc aacaaccaga tgctggtcga taaattccac
     4021 accatccgcg aaatcaccgt gccggcgtgg aagcggcaat tcatgctggc cttgagcctc
     4081 aacgagcaga agaacgccgt cgaactggcc acggccatcg acgacaccac caacgacctg
     4141 atgaagcgca atgcggccct gctgcatcgc acgtccgtcg agacggcgaa ggagaaccaa
     4201 cgcctggtga tcgacgtgga cacgctcaag caggttcaga cgacgctcat caagaccgtc
     4261 gaggacgtta ttcgcatcca gcaggaaggc gtgcagaagc gcaaggatgc cgagaagcag
     4321 atcgccgcaa tgcgtggcga tcttcaagcc aagctgaccc gccagcccgt gcgcgagctg
     4381 gcccaacagg agtccgtatg aatgccacaa acaccgatgt tttcgcccag gtaggcggcc
     4441 tggaggcccg aggcgcgaag atgaagaagc ggggcacccg cttcctcatc gcggcgctgg
     4501 cagtccttgc cattgccggg atcggggcag taacgggatg ggcgatcagc ccgagcgcga
     4561 cgcccggaag cattgacgtg ccgcaggtgc tggcatcgac attcagcgac caggtgccgg
     4621 gcagtgaggg cggcggcctg ggtggcggcc tgcccttcac ttcggccgtc ggggcattca
     4681 cggacttcat ggcggggccg gcaattttta ccttgggcat tcttggcata gtggtcgcgg
     4741 gtgccgtgct cgtgttcggg ggtgagttct gcgggttcgt gcgatccgtc tgcatgatgg
     4801 tgatagccgt cagcatgatt ttcgtgtcgt cgaacttggt gaagggcatt ctcggcggcg
     4861 atcacgacgc cggccctgcg gagccttcgc cgcgtgcgcg attcatggcg gccgtggagg
     4921 ccaaggattt cgcgcgagtg caagagctga tcgaggcgcg tggagccaag tcggcggctg
     4981 attatgtcct tgcgcagctc gccgtggccg aaggtctgga ccgcaagcct ggtgcgcgcg
     5041 tcgtggtcgg gaaagcggcg ggcagcatgg caatgccgcc tgcggcgctg ggttttacgc
     5101 caaggggaga agcggcatac gccatcgagc ggtcagccta tggtgagccg aggtccagca
     5161 ttgcgaagca gtaccagcag gaatggaacc ggaaggcggc gacctggtgg gcgatggccg
     5221 gtgtggccgg catcatcggc gcgatcctgg cggcggcggc aaccggcttt gttgggctgg
     5281 cagtgtcgat ccgcaaccga gtgaagcgcg tgcgcgacct gttggtgatg gagccgggtg
     5341 cagagccata agggatcgat ccccctcaag tcaaaagcct ccggtcggag gcttttgact
     5401 ttctgctcga gctaattcca ttgagtaagt ttttaagcac atcagcttca aaagcgctct
     5461 gaagttccta tactttctag agaataggaa cttcggaata ggaacttcaa gatccccaat
     5521 tcgagctcgg tacccgggga tcctagagtc gacctgcagg catgcaagct tggcactggc
     5581 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     5641 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     5701 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca
     5761 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     5821 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     5881 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     5941 gttttcaccg tcatcaccga aacgcgcga
//
