LOCUS       Exported                4789 bp ds-DNA    circular SYN 24-12-2013
DEFINITION  Plasmid with a kanMX marker for adding a C-terminal 12Pk tag.
ACCESSION   .
VERSION     .
KEYWORDS    pFA6a-12Pk-kanMX6
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4789)
  AUTHORS   Gadaleta MC, Iwasaki O, Noguchi C, Noma K, Noguchi E.
  TITLE     New vectors for epitope tagging and gene disruption in 
            Schizosaccharomyces pombe.
  JOURNAL   BioTechniques 2013;55:257-63.
  PUBMED    24215641
REFERENCE   2  (bases 1 to 4789)
  AUTHORS   Noguchi Lab
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4789
                     /organism="synthetic DNA construct"
                     /lab_host="Schizosaccharomyces pombe"
                     /mol_type="other DNA"
     CDS             73..672
                     /codon_start=1
                     /product="12 tandem copies of the Pk (V5) epitope"
                     /note="12Pk tag"
                     /translation="GKPIPNPLLGLDSTSGGKPIPNPLLGLDSTLDGGGKPIPNPLLGL
                     DSTSGGKPIPNPLLGLDSTLDGGGKPIPNPLLGLDSTSGGKPIPNPLLGLDSTLDGGGK
                     PIPNPLLGLDSTSGGKPIPNPLLGLDSTLDGGGKPIPNPLLGLDSTSGGKPIPNPLLGL
                     DSTLDGGGKPIPNPLLGLDSTSGGKPIPNPLLGLDST"
     terminator      702..889
                     /gene="S. cerevisiae ADH1"
                     /note="ADH1 terminator"
                     /note="transcription terminator for alcohol dehydrogenase 
                     1"
     gene            964..2320
                     /note="kanMX"
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        964..1307
                     /note="TEF promoter"
                     /note="Ashbya gossypii TEF promoter"
     CDS             1308..2117
                     /codon_start=1
                     /gene="aph(3)-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      2123..2320
                     /note="TEF terminator"
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(2425..2443)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2701..3289)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3460..4320)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4321..4425)
                     /gene="bla"
                     /note="AmpR promoter"
     promoter        4771..4789
                     /note="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"