LOCUS       Exported                9211 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pYZ41N, complete sequence.
ACCESSION   AF064066
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9211)
  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 9211)
  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 9211)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9211
                     /organism="Expression vector pYZ41N"
                     /host="Schizosaccharomyces pombe"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:76638"
     regulatory      17..1206
                     /regulatory_class="promoter"
                     /note="mutated; from Schizosaccharomyces pombe nmt1"
     regulatory      1108..1112
                     /regulatory_class="TATA_box"
                     /note="4 base deletion"
     CDS             complement(1201..1443)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWHMI"
     misc_feature    1207..1652
                     /label=from Escherichia coli lac operon
                     /note="from Escherichia coli lac operon"
     CDS             complement(1207..1443)
                     /codon_start=1
                     /product="lacZ alpha peptide"
                     /label=lacZ alpha peptide
                     /protein_id="AAC78828.1"
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWHM"
     primer_bind     1353..1369
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1370..1426
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1439..1455)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1463..1479
                     /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(1487..1517)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1532..1553
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      1652..2659
                     /regulatory_class="terminator"
                     /note="from Schizosaccharomyces pombe nmt1"
     rep_origin      2659..3859
                     /label=Schizosaccharomyces pombe ars1
                     /note="Schizosaccharomyces pombe ars1"
     rep_origin      2732..3515
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     misc_feature    3862..6969
                     /note="pUC119 sequence; contains ampicillin resistance 
                     gene"
     primer_bind     complement(3865..3881)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4094..4549
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4831..4935
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4936..5796
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AAC78829.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4936..5796
                     /gene="bla"
                     /label=bla
     rep_origin      5967..6555
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6843..6864
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6879..6909
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6917..6933
                     /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     6941..6957
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    6970..9211
                     /note="contains Saccharomyces cerevisiae leucine 
                     biosynthesis gene LEU2"
     CDS             complement(7458..8552)
                     /codon_start=1
                     /gene="LEU2"
                     /product="beta-isopropylmalate dehydrogenase"
                     /function="involved in leucine biosynthesis"
                     /label=LEU2
                     /protein_id="AAC78830.1"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     gene            complement(7458..8552)
                     /gene="LEU2"
                     /label=LEU2
     promoter        complement(8565..8969)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter