LOCUS       Exported                5444 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Integration vector pPyr-Kan, complete sequence.
ACCESSION   AY464559
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5444)
  AUTHORS   Middleton R, Hofmeister A.
  TITLE     New shuttle vectors for ectopic insertion of genes into Bacillus 
            subtilis
  JOURNAL   Plasmid 51 (3), 238-245 (2004)
  PUBMED    15109830
REFERENCE   2  (bases 1 to 5444)
  AUTHORS   Middleton RH, Hofmeister AEM.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-NOV-2003) Plant and Microbial Biology, University of 
            California, Berkeley, 281 Koshland Hall, Berkeley, CA 94704, USA
REFERENCE   3  (bases 1 to 5444)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5444
                     /organism="Integration vector pPyr-Kan"
                     /lab_host="Bacillus subtilis"
                     /mol_type="other DNA"
                     /note="origin of replication and ampicillin resistance for 
                     use in Escherichia coli, integrates into the chromosome of 
                     Bacillus subtilis at the pyrD locus; vector pieced together
                     from pUC18, pER82, and parts of the pyrD gene of Bacillus 
                     subtilis"
                     /db_xref="taxon:257526"
     misc_feature    complement(54..495)
                     /note="3 end of dihydroorotate dehydrogenase (catalytic 
                     subunit) (pyrD) of Bacillus subtilis"
     CDS             1467..2261
                     /codon_start=1
                     /gene="APHA"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=APHA
                     /note="kanamycin resistance"
                     /protein_id="AAR30046.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            1467..2261
                     /gene="APHA"
                     /label=APHA
     misc_feature    complement(2714..3133)
                     /note="5 end of dihydroorotate dehydrogenase (pyrD) of 
                     Bacillus subtilis"
     promoter        3538..3642
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3643..4503
                     /codon_start=1
                     /gene="BLA"
                     /product="beta-lactamase"
                     /label=BLA
                     /note="ampicillin resistance"
                     /protein_id="AAR30047.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            3643..4503
                     /gene="BLA"
                     /label=BLA
     rep_origin      4674..5262
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"