LOCUS       Exported                4824 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  PCR template vector pJJH727, complete sequence.
ACCESSION   AF298792
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4824)
  AUTHORS   Gueldener U, Heinisch J, Koehler GJ, Voss D, Hegemann JH.
  TITLE     A second set of loxP marker cassettes for Cre-mediated multiple gene
            knockouts in budding yeast
  JOURNAL   Nucleic Acids Res. 30 (6), E23 (2002)
  PUBMED    11884642
REFERENCE   2  (bases 1 to 4824)
  AUTHORS   Gueldener U, Heinisch JH, Voss D, Koehler G, Hegemann JH.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-AUG-2000) Institut fuer Mikrobiologie, 
            Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf 
            40225, Germany
REFERENCE   3  (bases 1 to 4824)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4824
                     /organism="PCR template vector pJJH727"
                     /mol_type="other DNA"
                     /label=derived from Saccharomyces cerevisiae
                     /note="derived from Saccharomyces cerevisiae"
                     /db_xref="taxon:142959"
     misc_feature    37..96
                     /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)."
     CDS             complement(240..1328)
                     /codon_start=1
                     /gene="LEU2"
                     /product="beta-isopropylmalate dehydrogenase"
                     /label=LEU2
                     /note="from Kluyveromyces lactis"
                     /protein_id="AAG34541.1"
                     /translation="MSKNIVVLPGDHVGKEVTDEAIKVLNAIAEVRPEIKFNFQHHLIG
                     GAAIDATGTPLPDEALEASKKADAVLLGAVGGPKWGTGAVRPEQGLLKIRKELGLYANL
                     RPCNFASDSLLDLSPLKPEYAKGTDFVVVRELVGGIYFGERKEDEGDGVAWDSEKYSVP
                     EVQRITRMAAFLALQQNPPLPIWSLDKANVLASSRLWRKTVEETIKTEFPQLTVQHQLI
                     DSAAMILVKSPTKLNGVVITNNMFGDIISDEASVIPGSLGLLPSASLASLPDTNKAFGL
                     YEPCHGSAPDLPANKVNPIATILSAAMMLKLSLDLVEEGRALEEAVRNVLDAGVRTGDL
                     GGSNSTTEVGDAIAKAVKEILA"
     gene            complement(240..1328)
                     /gene="LEU2"
                     /label=LEU2
     misc_feature    2359..2412
                     /label=loxP site
                     /note="loxP site"
     protein_bind    complement(2375..2408)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        complement(2462..2480)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2738..3326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3497..4357)
                     /codon_start=1
                     /gene="AmpR"
                     /product="ampicillin resistance protein"
                     /label=AmpR
                     /protein_id="AAG34542.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3497..4357)
                     /gene="AmpR"
                     /label=AmpR
     promoter        complement(4358..4462)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4808..2
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"