LOCUS       Exported                9956 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pYZ3N-GFP, complete sequence.
ACCESSION   AF049064
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9956)
  AUTHORS   Zhao Y, Elder RT, Chen M.
  TITLE     Fission yeast expression vector with GFP tag and positive selection 
            for insert
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9956)
  AUTHORS   Zhao Y, Chen M, Elder RT, Cao J.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-FEB-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 9956)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9956
                     /organism="Expression vector pYZ3N-GFP"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:73425"
     regulatory      17..1213
                     /regulatory_class="promoter"
                     /note="from Schizosaccharomyces pombe nmt1"
     gene            1125..1938
                     /gene="GFP"
                     /label=GFP
     CDS             1225..1938
                     /codon_start=1
                     /gene="GFP"
                     /product="green fluorescent protein"
                     /label=GFP
                     /note="bright version of GFP (P. Nurse, personal 
                     communication); M2GFP: S65A, V68L, S72A"
                     /protein_id="AAC78774.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             complement(1930..2184)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRGRGTFV"
     misc_feature    complement(1945..2394)
                     /note="Escherichia coli lac operon; contains multiple 
                     cloning sites"
     CDS             complement(1945..2184)
                     /codon_start=1
                     /product="lacZ alpha peptide"
                     /label=lacZ alpha peptide
                     /protein_id="AAC78775.1"
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRGR"
     primer_bind     2094..2110
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2111..2167
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2180..2196)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2204..2220
                     /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(2228..2258)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2273..2294
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      2394..3403
                     /regulatory_class="terminator"
                     /note="from Schizosaccharomyces pombe nmt1"
     rep_origin      3404..4604
                     /label=Schizosaccharomyces pombe ars1
                     /note="Schizosaccharomyces pombe ars1"
     rep_origin      3477..4260
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     misc_feature    4605..7714
                     /note="pUC119 sequence; contains ampicillin resistance 
                     gene"
     primer_bind     complement(4610..4626)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4839..5294
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5576..5680
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5681..6541
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AAC78776.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            5681..6541
                     /gene="bla"
                     /label=bla
                     /note="ampicillin resistance gene"
     rep_origin      6712..7300
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    7588..7609
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7624..7654
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7662..7678
                     /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     7686..7702
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    7715..9956
                     /note="Saccharomyces cerevisiae leucine biosynthesis gene 
                     LEU2"
     CDS             complement(8203..9297)
                     /codon_start=1
                     /gene="Leu2"
                     /product="beta-isopropylmalate dehydrogenase"
                     /function="involved in leucine biosynthesis"
                     /label=Leu2
                     /protein_id="AAC78777.1"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     gene            complement(8203..9297)
                     /gene="Leu2"
                     /label=Leu2
     promoter        complement(9310..9714)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter