LOCUS Exported 3377 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pMWO-078, complete sequence.
ACCESSION JQ283950
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3377)
AUTHORS Obrist MW, Miller VL.
TITLE Low copy expression vectors for use in Yersinia sp. and related
organisms
JOURNAL Plasmid 68 (1), 33-42 (2012)
PUBMED 22445322
REFERENCE 2 (bases 1 to 3377)
AUTHORS Obrist MW, Miller VL.
TITLE Direct Submission
JOURNAL Submitted (14-DEC-2011) Microbiology and Immunology, University of
North Carolina at Chapel Hill, 116 Manning Drive, 505 Mary Ellen
Jones, CB# 7290, Chapel Hill, NC 27599, USA
REFERENCE 3 (bases 1 to 3377)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Assembly Method :: DNASTAR Lasergene SeqMan Pro v. 7.2.1 (1)
Intel
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..3377
/organism="Expression vector pMWO-078"
/mol_type="other DNA"
/db_xref="taxon:1166520"
primer_bind 60..77
/note="MWO-fw; forward sequencing primer"
promoter 75..148
/label=PLtetO-1 promoter
/note="modified phage lambda PL promoter with tet operator
sites (Lutz and Bujard, 1997)"
protein_bind 75..93
/label=tet operator 2 binding site
/bound_moiety="tet operator 2"
/note="operator 2 from Tn10; tet repressor binding site"
regulatory 94..99
/regulatory_class="minus_35_signal"
protein_bind 100..118
/label=tet operator 2 binding site
/bound_moiety="tet operator 2"
/note="operator 2 from Tn10; tet repressor binding site"
regulatory 117..122
/regulatory_class="minus_10_signal"
misc_feature 155..297
/label=multiple cloning site from pBluescript II KS+
/note="multiple cloning site from pBluescript II KS+"
misc_feature 155..262
/label=MCS
/note="pBluescript multiple cloning site"
primer_bind 172..188
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(222..238)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(271..289)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 298..1202
/label=region containing p15A origin of replication
/note="region containing p15A origin of replication"
primer_bind complement(384..405)
/note="MWO-rv-2; reverse sequencing primer"
rep_origin complement(557..1102)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin."
terminator 1245..1331
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
regulatory 1285..1328
/regulatory_class="terminator"
/note="rrnB T1"
CDS complement(1382..2005)
/codon_start=1
/gene="tetR"
/product="tetracycline repressor protein class B"
/label=tetR
/note="from transposon Tn10"
/protein_id="AFF57862.1"
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNKAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
gene complement(1382..2005)
/gene="tetR"
/label=tetR
CDS complement(2222..3013)
/codon_start=1
/gene="aadA"
/product="streptomycin 3-adenylyltransferase"
/label=aadA
/protein_id="AFF57863.1"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
gene complement(2222..3013)
/gene="aadA"
/label=aadA
LOCUS Exported 3377 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pMWO-078, complete sequence.
ACCESSION JQ283950
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3377)
AUTHORS Obrist MW, Miller VL.
TITLE Low copy expression vectors for use in Yersinia sp. and related
organisms
JOURNAL Plasmid 68 (1), 33-42 (2012)
PUBMED 22445322
REFERENCE 2 (bases 1 to 3377)
AUTHORS Obrist MW, Miller VL.
TITLE Direct Submission
JOURNAL Submitted (14-DEC-2011) Microbiology and Immunology, University of
North Carolina at Chapel Hill, 116 Manning Drive, 505 Mary Ellen
Jones, CB# 7290, Chapel Hill, NC 27599, USA
REFERENCE 3 (bases 1 to 3377)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Assembly Method :: DNASTAR Lasergene SeqMan Pro v. 7.2.1 (1)
Intel
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..3377
/organism="Expression vector pMWO-078"
/mol_type="other DNA"
/db_xref="taxon:1166520"
primer_bind 60..77
/note="MWO-fw; forward sequencing primer"
promoter 75..148
/label=PLtetO-1 promoter
/note="modified phage lambda PL promoter with tet operator
sites (Lutz and Bujard, 1997)"
protein_bind 75..93
/label=tet operator 2 binding site
/bound_moiety="tet operator 2"
/note="operator 2 from Tn10; tet repressor binding site"
regulatory 94..99
/regulatory_class="minus_35_signal"
protein_bind 100..118
/label=tet operator 2 binding site
/bound_moiety="tet operator 2"
/note="operator 2 from Tn10; tet repressor binding site"
regulatory 117..122
/regulatory_class="minus_10_signal"
misc_feature 155..297
/label=multiple cloning site from pBluescript II KS+
/note="multiple cloning site from pBluescript II KS+"
misc_feature 155..262
/label=MCS
/note="pBluescript multiple cloning site"
primer_bind 172..188
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(222..238)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(271..289)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 298..1202
/label=region containing p15A origin of replication
/note="region containing p15A origin of replication"
primer_bind complement(384..405)
/note="MWO-rv-2; reverse sequencing primer"
rep_origin complement(557..1102)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin."
terminator 1245..1331
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
regulatory 1285..1328
/regulatory_class="terminator"
/note="rrnB T1"
CDS complement(1382..2005)
/codon_start=1
/gene="tetR"
/product="tetracycline repressor protein class B"
/label=tetR
/note="from transposon Tn10"
/protein_id="AFF57862.1"
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNKAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
gene complement(1382..2005)
/gene="tetR"
/label=tetR
CDS complement(2222..3013)
/codon_start=1
/gene="aadA"
/product="streptomycin 3-adenylyltransferase"
/label=aadA
/protein_id="AFF57863.1"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
gene complement(2222..3013)
/gene="aadA"
/label=aadA