LOCUS       Exported               10231 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast expression vector pSZPT1 DNA, complete sequence.
ACCESSION   AB871644
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10231)
  AUTHORS   Kanda K, Ishida T, Hirota R, Ono S, Motomura K, Ikeda T, Kitamura K,
            Kuroda A.
  TITLE     Application of a phosphite dehydrogenase gene as a novel dominant 
            selection marker for yeasts
  JOURNAL   J. Biotechnol. 182-183, 68-73 (2014)
  PUBMED    24786825
REFERENCE   2  (bases 1 to 10231)
  AUTHORS   Hirota R.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-NOV-2013) Contact:Ryuichi Hirota Hiroshima University,
            Department of Biotechnology; 1-3-1, Higashi-Hiroshima, Hiroshima 
            739-8530, Japan
REFERENCE   3  (bases 1 to 10231)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10231
                     /organism="Yeast expression vector pSZPT1"
                     /focus
                     /mol_type="other DNA"
                     /db_xref="taxon:1430310"
     source          1251..2261
                     /organism="Ralstonia sp. 4506"
                     /strain="4506"
                     /mol_type="other DNA"
                     /country="Japan"
                     /isolation_source="soil"
                     /collected_by="Hirota,R."
                     /collection_date=2011
                     /db_xref="taxon:1094341"
     misc_feature    7..12
                     /label=SphI recognition site
                     /note="SphI recognition site"
     misc_feature    13..18
                     /label=PstI recognition site
                     /note="PstI recognition site"
     regulatory      36..1204
                     /regulatory_class="promoter"
                     /note="nmt1 promoter"
     misc_feature    1211..1216
                     /label=SalI recognition site
                     /note="SalI recognition site"
     CDS             1233..1250
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1251..2261
                     /codon_start=1
                     /gene="RsptxD"
                     /product="phosphite dehydrogenase"
                     /label=RsptxD
                     /protein_id="BAP74302.1"
                     /translation="MKPKVVLTHWVHPEIIELLSASADVIPNTTRETLPRSEVIARAKD
                     ADALMAFMPDSIDSAFLEECPKLRVIGAALKGYDNFDVNACTRHGVWLTIVPDLLTIPT
                     AELTIGLLLGLTRHMLEGDRQIRSGHFQGWRPTLYGSGLTGKTLGIIGMGAVGRAIAQR
                     LAGFEMNLLYCDPIPLNAEQEKAWHVQRVTLDELLEKCDYVVPMVPMAAETLHLIDATA
                     LAKMKTGSYLINACRGSVVDENAVIAALASGKLAGYAADVFEMEEWIRADRPQAIPKAL
                     LDNTAQTFFTPHLGSAVKEVRLEIERQAAMNIIQALAGEKPMGAINQPYPGVKAA"
     gene            1251..2261
                     /gene="RsptxD"
                     /label=RsptxD
     misc_feature    2262..1250
                     /label=derived from pDUAL-HFF1(AB364046.1)
                     /note="derived from pDUAL-HFF1(AB364046.1)"
     CDS             2262..2285
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2292..2315
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      2345..2473
                     /regulatory_class="terminator"
                     /note="ADH1 terminator"
     terminator      2367..2554
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     misc_feature    2584..2589
                     /label=SpeI recognition site
                     /note="SpeI recognition site"
     misc_feature    2590..3816
                     /label=leu1-5
                     /note="leu1-5"
     CDS             complement(4218..5012)
                     /codon_start=1
                     /gene="ura4"
                     /product="orotidine 5-phosphate decarboxylase"
                     /label=ura4
                     /protein_id="BAP74303.1"
                     /translation="MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKS
                     EILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVK
                     LQYASGVYKIASWAHITNCHTVPGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSY
                     TEKTLEWFEKHTDFCFGFIAGRRFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIV
                     NCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH"
     gene            complement(4218..5012)
                     /gene="ura4"
                     /label=ura4
     misc_feature    5548..5553
                     /label=EcoRI recognition site
                     /note="EcoRI recognition site"
     rep_origin      5554..6746
                     /label=ars1
                     /note="ars1"
     rep_origin      5621..6404
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     misc_feature    6761..7114
                     /label=leu1-3
                     /note="leu1-3"
     misc_feature    7115..7120
                     /label=SacI recognition site
                     /note="SacI recognition site"
     misc_feature    7121..7126
                     /label=EcoRI recognition site
                     /note="EcoRI recognition site"
     primer_bind     complement(7127..7143)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      7356..7811
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        8093..8197
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8198..9058
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      9229..9817
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    10105..10126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        10141..10171
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10179..10195
                     /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)."
     primer_bind     10203..10219
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"