LOCUS       Exported               10804 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  S. pombe expression vector pDUAL2-HFC41c DNA, complete sequence.
ACCESSION   AB364108
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10804)
  AUTHORS   Matsuyama A, Shirai A, Yashiroda Y, Kamata A, Horinouchi S, Yoshida 
            M.
  TITLE     pDUAL, a multipurpose, multicopy vector capable of chromosomal 
            integration in fission yeast
  JOURNAL   Yeast 21 (15), 1289-1305 (2004)
  PUBMED    15546162
REFERENCE   2  (bases 1 to 10804)
  AUTHORS   Matsuyama A, Shirai A, Yoshida M.
  TITLE     Fission yeast multicopy and integration vector pDUAL2-HFC41c
  JOURNAL   Published Only in Database (2007)
REFERENCE   3  (bases 1 to 10804)
  AUTHORS   Matsuyama A, Shirai A, Yoshida M.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-OCT-2007) Akihisa Matsuyama, RIKEN, Chemical Genetics 
            Laboratory; Hirosawa 2-1, Wako, Saitama 351-0198, Japan 
            (E-mail:akihisa@riken.jp, URL:http://cgl.riken.go.jp/, 
            Tel:81-48-462-1335, Fax:81-48-462-1336)
REFERENCE   4  (bases 1 to 10804)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     Fission yeast expression vector.
FEATURES             Location/Qualifiers
     source          1..10804
                     /organism="Schizosaccharomyces pombe expression vector 
                     pDUAL2-HFC41c"
                     /mol_type="other DNA"
                     /db_xref="taxon:477386"
     regulatory      1..237
                     /regulatory_class="terminator"
                     /gene="ADH1"
     misc_feature    1..237
                     /gene="ADH1"
                     /label=derived from Saccharomyces cerevisiae
                     /note="derived from Saccharomyces cerevisiae"
     gene            1..237
                     /gene="ADH1"
                     /label=ADH1
     terminator      21..208
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     misc_feature    242..2193
                     /label=derived from Schizosaccharomyces pombe
                     /note="derived from Schizosaccharomyces pombe"
     CDS             join(242..618,1840..2193)
                     /pseudo
                     /gene="leu1"
                     /label=leu1
     gene            join(242..618,1840..2193)
                     /pseudo
                     /gene="leu1"
                     /label=leu1
     rep_origin      633..1825
                     /standard_name="ars1"
                     /label=ars1
     rep_origin      975..1758
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     misc_feature    2194..5316
                     /label=derived from Cloning vector pUC119
                     /note="derived from Cloning vector pUC119"
     primer_bind     complement(2206..2222)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2435..2890
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3172..3276
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3277..4137
                     /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      4308..4896
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5184..5205
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5220..5250
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5258..5274
                     /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     5282..5298
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5317..8269
                     /label=derived from Schizosaccharomyces pombe
                     /note="derived from Schizosaccharomyces pombe"
     gene            complement(5317..7068)
                     /gene="ura4"
                     /label=ura4
     regulatory      complement(5317..5746)
                     /regulatory_class="terminator"
                     /gene="ura4"
     CDS             complement(5747..6541)
                     /codon_start=1
                     /gene="ura4"
                     /product="orotidine 5-phosphate decarboxylase"
                     /label=ura4
                     /note="internal EcoRV site is eliminated."
                     /protein_id="BAF90877.1"
                     /translation="MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKS
                     EILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVK
                     LQYASGVYKIASWAHITNCHTVPGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSY
                     TEKTLEWFEKHTDFCFGFIAGRRFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIV
                     NCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH"
     regulatory      complement(6542..7068)
                     /regulatory_class="promoter"
                     /gene="ura4"
     regulatory      7104..8269
                     /regulatory_class="promoter"
                     /gene="nmt1"
                     /note="intermediate strength nmt1 promoter"
     gene            7104..8269
                     /gene="nmt1"
                     /label=nmt1
     CDS             8295..8342
                     /codon_start=1
                     /product="hexahistidine and the FLAG epitope"
                     /label=hexahistidine and the FLAG epitope
                     /protein_id="BAF90878.1"
                     /translation="MHHHHHHMDYKDDDDK"
     CDS             8298..8315
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             8319..8342
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8346..9062
                     /codon_start=1
                     /product="cyan fluorescent protein"
                     /label=cyan fluorescent protein
                     /protein_id="BAF90879.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    9066..10776
                     /standard_name="ccdB cassette reading frame A"
                     /label=ccdB cassette reading frame A
     protein_bind    9069..9193
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        9218..9248
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             9302..9961
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             10303..10608
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(10649..10773)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"