LOCUS       Exported                7500 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pRS415, complete sequence.
ACCESSION   FJ012804
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7500)
  AUTHORS   Pinto MR, Marchini JF, Buranello PA, Oliver C, Mortara RA, Tosi LR.
  TITLE     The Leishmania major HTBF protein belongs in the YIP family of 
            proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7500)
  AUTHORS   Pinto MR, Marchini JF, Tosi LR.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-AUG-2008) Bilogia Celular e Molecular e Bioagentes 
            Patogenicos, Faculdade de Medicina de Ribeirao Preto, AV. 
            Bandeirantes, 3900, Ribeirao Preto, Sao Paulo 14049-900, Brasil
REFERENCE   3  (bases 1 to 7500)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7500
                     /organism="Shuttle vector pRS415"
                     /mol_type="other DNA"
                     /note="BamHI/Xmai digestion of pRS415 ligated to BamHi/Xmai
                     digestion of (5- gcgcccgggcatatgctcaacgaggtgc/5- 
                     cgcggatcctaaataccaaccaga) PCR amplification of pELHYG; T1 
                     product was digested with HindIII/Xmai and ligated to 931bp
                     HindIII/Xmai fragment of pRS316"
                     /db_xref="taxon:554026"
     CDS             complement(663..1757)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1770..2174)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2474..2929)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     3074..3090
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3100..3118
                     /label=T7
                     /note="T7"
     promoter        3100..3118
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3144..3160
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(3181..3267)
                     /regulatory_class="promoter"
                     /note="GAL 1"
     promoter        3272..3936
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    3603..3720
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     regulatory      3948..4088
                     /regulatory_class="promoter"
                     /note="GAL10"
     CDS             4103..4672
                     /codon_start=1
                     /product="HTBF"
                     /label=HTBF
                     /note="H region resistance protein; derived from Leishmania
                     terbinafine"
                     /protein_id="ACH69155.1"
                     /translation="MLNEVHVNADPNSISTLDEPVLETLLRDAKAIGRKLVVVVCPSLG
                     GDKELRDWDLWGPLFLCLILASILTINASDDQGAAVFSAVFIFVWLGGLVVTVNAKLLG
                     SKIMFFQTYCAMGYCLAPICLGALLCCVLPWFLLNLMLCFMAWAWACWAALRFFRHTVS
                     ADREVLVVYPVGLFYVFFTWMVLVGI"
     primer_bind     complement(4673..4689)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4726..4744)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4765..4781)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4789..4805
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4813..4843)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(5167..5755)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5926..6786)
                     /codon_start=1
                     /product="ampicillin resistance protein"
                     /label=ampicillin resistance protein
                     /protein_id="ACH69156.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6787..6891)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6928..7431
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"