LOCUS       Exported                5623 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Tagging vector pFA6a-hphMX6-P81nmt1-3FLAG, complete sequence.
ACCESSION   EU689034
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5623)
  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 5623)
  AUTHORS   Noguchi C, Garabedian M, Malik M, Noguchi E.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-MAY-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 5623)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5623
                     /organism="Tagging vector pFA6a-hphMX6-P81nmt1-3FLAG"
                     /mol_type="other DNA"
                     /db_xref="taxon:554660"
     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="ACH53402.1"
                     /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..4045)
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     terminator      2470..2667
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(2673..3701)
                     /codon_start=1
                     /gene="hph"
                     /product="hygromycin-B-phosphotransferase"
                     /label=hph
                     /note="hygromycin resistance protein"
                     /protein_id="ACH53403.1"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     gene            complement(2673..3701)
                     /gene="hph"
                     /label=hph
     promoter        complement(3702..4045)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     regulatory      4086..5258
                     /regulatory_class="promoter"
                     /note="nmt1 promoter P81"
     promoter        4090..5248
                     /label=nmt1 P81 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring weak thiamine-repressible expression"
     CDS             5263..5340
                     /codon_start=1
                     /product="3xFLAG"
                     /label=3xFLAG
                     /protein_id="ACH53404.1"
                     /translation="MDYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             5266..5337
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     terminator      5361..5548
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"