LOCUS       Exported                9208 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pYZ81N, complete sequence.
ACCESSION   AF064067
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9208)
  AUTHORS   Zhao Y, Elder RT, Chen M, Cao J.
  TITLE     Fission yeast expression vectors adapted for positive identification
            of gene insertion and green fluorescent protein fusion
  JOURNAL   BioTechniques 25 (3), 438-440 (1998)
  PUBMED    9762442
REFERENCE   2  (bases 1 to 9208)
  AUTHORS   Zhao Y, Chen M.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-MAY-1998) Departments of Pediatrics and 
            Microbiology-Immunology, Northwestern University Medical School, 
            2430 N Halsted Street, #218, Chicago, IL 60614, USA
REFERENCE   3  (bases 1 to 9208)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9208
                     /organism="Expression vector pYZ81N"
                     /host="Schizosaccharomyces pombe"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:76639"
     regulatory      17..1203
                     /regulatory_class="promoter"
                     /note="mutated; from Schizosaccharomyces pombe nmt1"
     regulatory      1108..1109
                     /regulatory_class="TATA_box"
                     /note="7 base deletion"
     CDS             complement(1198..1440)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWHMI"
     misc_feature    1204..1648
                     /label=from Escherichia coli lac operon
                     /note="from Escherichia coli lac operon"
     CDS             complement(1204..1440)
                     /codon_start=1
                     /product="lacZ alpha peptide"
                     /label=lacZ alpha peptide
                     /protein_id="AAC78831.1"
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWHM"
     primer_bind     1350..1366
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1367..1423
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1436..1452)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1460..1476
                     /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(1484..1514)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1529..1550
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      1649..2656
                     /regulatory_class="terminator"
                     /note="from Schizosaccharomyces pombe nmt1"
     rep_origin      2656..3856
                     /label=Schizosaccharomyces pombe ars1
                     /note="Schizosaccharomyces pombe ars1"
     rep_origin      2729..3512
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     misc_feature    3858..6966
                     /note="pUC119 sequence; contains ampicillin resistance 
                     gene"
     primer_bind     complement(3862..3878)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4091..4546
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4828..4932
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4933..5793
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AAC78832.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4933..5793
                     /gene="bla"
                     /label=bla
     rep_origin      5964..6552
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6840..6861
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6876..6906
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6914..6930
                     /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     6938..6954
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    6967..9208
                     /note="contains Saccharomyces cerevisiae leucine 
                     biosynthesis gene LEU2"
     CDS             complement(7455..8549)
                     /codon_start=1
                     /gene="LEU2"
                     /product="beta-isopropylmalate dehydrogenase"
                     /function="involved in leucine biosynthesis"
                     /label=LEU2
                     /protein_id="AAC78833.1"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     gene            complement(7455..8549)
                     /gene="LEU2"
                     /label=LEU2
     promoter        complement(8562..8966)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter