LOCUS       Exported                5411 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Tagging vector pFA6a-KanMX6-P3nmt1-3FLAG, complete sequence.
ACCESSION   EU493331
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5411)
  AUTHORS   Noguchi C, Garabedian MV, Malik M, Noguchi E.
  TITLE     A vector system for genomic FLAG epitope-tagging in 
            Schizosaccharomyces pombe
  JOURNAL   Biotechnol J 3 (9-10), 1280-1285 (2008)
  PUBMED    18729046
REFERENCE   2  (bases 1 to 5411)
  AUTHORS   Noguchi C, Garabedian M, Malik M, Noguchi E.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-FEB-2008) Biochemistry and Molecular Biology, Drexel 
            University College of Medicine, 245 N 15th Street, MS 497, 
            Philadelphia, PA 19102, USA
REFERENCE   3  (bases 1 to 5411)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5411
                     /organism="Tagging vector pFA6a-KanMX6-P3nmt1-3FLAG"
                     /mol_type="other DNA"
                     /label=pFA6a derivative
                     /note="pFA6a derivative"
                     /db_xref="taxon:524041"
     promoter        complement(1..19)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        365..469
                     /gene="bla"
                     /label=AmpR promoter
     CDS             470..1330
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="ACD03182.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            470..1330
                     /gene="bla"
                     /label=bla
     rep_origin      1501..2089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        2347..2365
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            complement(2470..3826)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      2470..2667
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(2673..3482)
                     /codon_start=1
                     /gene="kanR"
                     /product="kanamycin and geneticin (G418) resistance 
                     protein"
                     /label=kanR
                     /protein_id="ACD03183.1"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(2673..3482)
                     /gene="kanR"
                     /label=kanR
     promoter        complement(3483..3826)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     misc_feature    3866..5045
                     /label=nmt1 promoter (P3)
                     /note="nmt1 promoter (P3)"
     promoter        3871..5036
                     /label=nmt1 promoter
                     /note="wild-type nmt1 promoter from Schizosaccharomyces 
                     pombe, conferring strong thiamine-repressible expression"
     CDS             5051..5128
                     /codon_start=1
                     /product="3 x FLAG"
                     /label=3 x FLAG
                     /protein_id="ACD03184.1"
                     /translation="MDYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             5054..5125
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     terminator      5149..5336
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"