LOCUS       Exported                9406 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Transposon delivery vector pAW068, complete sequence.
ACCESSION   EU146228
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9406)
  AUTHORS   Wilson AC, Perego M, Hoch JA.
  TITLE     New transposon delivery plasmids for insertional mutagenesis in 
            Bacillus anthracis
  JOURNAL   J. Microbiol. Methods 71 (3), 332-335 (2007)
  PUBMED    17931726
REFERENCE   2  (bases 1 to 9406)
  AUTHORS   Wilson AC, Perego M, Hoch JA.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-SEP-2007) Department of Molecular and Experimental, 
            The Scripps Research Institute, 10550 North Torrey Pines Road, La 
            Jolla, CA 92037, USA
REFERENCE   3  (bases 1 to 9406)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9406
                     /organism="Transposon delivery vector pAW068"
                     /mol_type="other DNA"
                     /db_xref="taxon:474006"
     CDS             171..1217
                     /codon_start=1
                     /product="C9 transposase"
                     /label=C9 transposase
                     /protein_id="ABV70027.1"
                     /translation="MEKKEFRVLIKYCFLKGKNTVEAKTWLDNEFPDSAPGKSTIIDWY
                     AKFKRGEMSTEDGERSGRPKEVVTDENIKKIHKMILNDRKMKLIEIAEALKISKERVGH
                     IIHQYLDMRKLCAKWVPRELTFDQKQRRVDDSKRCLQLLTRNTPEFFRRYVTMDETWLH
                     HYTPESNRQSAEWTATGEPSPKRGKTQKSAGKVMASVFWDAHGIIFIDYLEKGKTINSD
                     YYMALLERLKVEIAAKRPHMKKKKVLFHQDNAPCHKSLRTMAKIHELGFELLPHPPYSP
                     DLAPSDFFLFSDLKRMLAGKKFGCNEEVIAETEAYFEAKPKEYYQNGIKKLEGRYNRCI
                     ALEGNYVE"
     promoter        complement(1280..1298)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1635..1656
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1671..1701
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1709..1725
                     /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     1733..1749
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1770..1788
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     mobile_element  1801..1827
                     /mobile_element_type="transposon:mariner"
     CDS             1843..1908
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    1912..1959
                     /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)."
     promoter        complement(1978..1996)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3367..4149
                     /codon_start=1
                     /product="spectinomycin resistance protein"
                     /label=spectinomycin resistance protein
                     /protein_id="ABV70028.1"
                     /translation="MTNLINGKIPNQAIQTLKIVKDLFGSSIVGVYLFGSAVNGGLRIN
                     SDVDGLVVVNHSLPQLTRKKLTERLMTISGKIGNTDSVRPLEVTVINRSEVVPWQYPPK
                     REFIYGEWLRGEFENGQIQEPSYDPDLAIVLAQARKNSISLFGPDSSSILVSVPLTDIR
                     RAIKDSLPELIEGIKGDERNVILTVAAMWQTVTTGEITSKDVAAEWAIPLLPKEHVTLL
                     DIARKGYRGEWYDKWEGLYSKVKALVKYMKNSIETSLN"
     protein_bind    5051..5098
                     /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)."
     mobile_element  complement(5112..5138)
                     /mobile_element_type="transposon:mariner"
     promoter        complement(5151..5169)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5176..5192)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5817..6467
                     /codon_start=1
                     /product="chloramphenicol resistance protein"
                     /label=chloramphenicol resistance protein
                     /protein_id="ABV70029.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     CDS             complement(7080..7778)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /protein_id="ABV70030.1"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNNSNIIQNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     gene            complement(7080..7778)
                     /gene="repA"
                     /label=repA
     CDS             8390..8401
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"