LOCUS       Exported                5969 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWFRT-tel, complete sequence.
ACCESSION   EU329006
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5969
                     /organism="Cloning vector pWFRT-tel"
                     /mol_type="other DNA"
                     /label=Burkholderia sp. suicidal vector
                     /note="Burkholderia sp. suicidal vector"
                     /db_xref="taxon:491085"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             201..1061
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="ABY57995.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            201..1061
                     /gene="bla"
                     /label=bla
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      1266..1884
                     /label=ColE1
                     /note="ColE1"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /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)."
     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
                     /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
     regulatory      5369..5400
                     /regulatory_class="terminator"
     terminator      5369..5400
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     misc_feature    5444..5517
                     /label=Flip recognition target
                     /note="Flip recognition target"
     protein_bind    complement(5461..5508)
                     /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)."
     primer_bind     complement(5575..5591)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"