LOCUS       Exported                5702 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Fungal transformation vector pKT312, complete sequence.
ACCESSION   MH286805
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5702)
  AUTHORS   Twaruschek K, Spoerhase P, Michlmayr H, Wiesenberger G, Adam G.
  TITLE     New plasmids allowing positive-negative selection for fungal 
            transformation and efficient Cre-loxP mediated marker recycling
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5702)
  AUTHORS   Twaruschek K, Spoerhase P, Michlmayr H, Wiesenberger G, Adam G.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-MAY-2018) Department of Applied Genetics and Cell 
            Biology, University of Natural Resources and Life Sciences Vienna 
            (BOKU), Konrad Lorenz Strasse 24, Tulln 3430, Austria
REFERENCE   3  (bases 1 to 5702)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5702
                     /organism="Fungal transformation vector pKT312"
                     /mol_type="other DNA"
                     /db_xref="taxon:2211131"
     regulatory      62..808
                     /regulatory_class="promoter"
                     /note="pki; Trichoderma reesei pyruvate kinase I promoter; 
                     accession CP016233"
     regulatory      215..224
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             814..1941
                     /codon_start=1
                     /gene="HSVtk"
                     /product="HSV thymidine kinase"
                     /label=HSVtk
                     /note="no stop codon due to C-terminal resistance gene 
                     fusion"
                     /protein_id="AWN56797.1"
                     /translation="MASYPCHQHASAFDQAARSRGHNNRRTALRPRRQQKATEVRLEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGAAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     gene            814..1941
                     /gene="HSVtk"
                     /label=HSVtk
     CDS             1954..2526
                     /codon_start=1
                     /gene="nat1"
                     /product="nourseothricin aminotransferase"
                     /label=nat1
                     /protein_id="AWN56798.1"
                     /translation="MTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFT
                     LREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVISYSAWNRR
                     LTVEDIEVAPEHRGHGVGRALMGLATEFAGERGAGHLWLEVTNVNAPAIHAYRRMGFTL
                     CGLDTALYDGTASDGERQALYMSMPCP"
     gene            1954..2526
                     /gene="nat1"
                     /label=nat1
     regulatory      2544..3273
                     /regulatory_class="terminator"
                     /note="cbh2; Trichoderma reesei cellobiohydrolase II 
                     terminator; accession CP016238"
     primer_bind     complement(3339..3358)
                     /label=T7 (for sequencing of downstream insert)
                     /note="T7 (for sequencing of downstream insert)"
     promoter        complement(3340..3358)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(3598..4280)
                     /direction=LEFT
                     /label=ColE1 origin
                     /note="ColE1 origin"
     rep_origin      complement(3616..4204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4375..5235)
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /note="ampicillin resistance protein"
                     /protein_id="AWN56796.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(4375..5235)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(5236..5340)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5686..2
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5686..5702)
                     /label=SP6 (for sequencing of upstream insert)
                     /note="SP6 (for sequencing of upstream insert)"