LOCUS       Exported                3760 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPL5618_pUG_MtxR, complete sequence.
ACCESSION   KP159512
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3760)
  AUTHORS   MacDonald C, Piper RC.
  TITLE     Puromycin- and methotrexate-resistance cassettes and optimized 
            Cre-recombinase expression plasmids for use in yeast
  JOURNAL   Yeast 32 (5), 423-438 (2015)
  PUBMED    25688547
REFERENCE   2  (bases 1 to 3760)
  AUTHORS   MacDonald C, Piper RC.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-NOV-2014) Molecular Physiology & Biophysics, 
            University of Iowa, 51 Newton Road, Iowa City, IA 52242, USA
REFERENCE   3  (bases 1 to 3760)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Sequence Builder (Lasergene) v. 8.0.3 
            Sequencing Technology :: Assembled
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3760
                     /organism="Cloning vector pPL5618_pUG_MtxR"
                     /mol_type="other DNA"
                     /db_xref="taxon:1711425"
     misc_feature    53..86
                     /label=loxP site
                     /note="loxP site"
     protein_bind    complement(53..86)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      97..484
                     /regulatory_class="promoter"
                     /note="from TEF1"
     promoter        141..484
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             485..1048
                     /codon_start=1
                     /gene="DHFR*"
                     /product="Dihydrofolate Reductase L22F F31S"
                     /label=DHFR*
                     /protein_id="ALD15651.1"
                     /translation="MVGSLNCIVAVSQNMGIGKNGDFPWPPLRNESRYFQRMTTTSSVE
                     GKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLSRSLDDALKLTEQP
                     ELANKVDMVWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEY
                     PGVLSDVQEEKGIKYKLEVYEKND"
     gene            485..1048
                     /gene="DHFR*"
                     /label=DHFR*
     regulatory      1053..1278
                     /regulatory_class="terminator"
                     /note="from TEF1"
     terminator      1054..1251
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_feature    1311..1344
                     /label=loxP site
                     /note="loxP site"
     protein_bind    complement(1311..1344)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        complement(1398..1416)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(1674..2262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2433..3293)
                     /codon_start=1
                     /gene="AmpR"
                     /product="Ampicillin resistance protein"
                     /label=AmpR
                     /protein_id="ALD15652.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2433..3293)
                     /gene="AmpR"
                     /label=AmpR
     promoter        complement(3294..3398)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3744..2
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"