LOCUS Exported 6610 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pTEV19, complete sequence.
ACCESSION KU974159
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6610)
AUTHORS VanDrisse CM, Escalante-Semerena JC.
TITLE New high-cloning-efficiency vectors for complementation studies and
recombinant protein overproduction in Escherichia coli and
Salmonella enterica
JOURNAL Plasmid (2016) In press
PUBMED 27234933
REFERENCE 2 (bases 1 to 6610)
AUTHORS VanDrisse CM, Escalante-Semerena JC.
TITLE Direct Submission
JOURNAL Submitted (24-MAR-2016) Microbiology, University of Georgia, 120
Cedar St., Athens, GA 30602, USA
REFERENCE 3 (bases 1 to 6610)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6610
/organism="Expression vector pTEV19"
/mol_type="other DNA"
/db_xref="taxon:1856676"
CDS 56..73
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 140..187
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 224..679
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 706..810
/gene="bla"
/label=AmpR promoter
CDS 811..1671
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=bla
/protein_id="ANI69980.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene 811..1671
/gene="bla"
/label=bla
rep_origin 1842..2430
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 2616..2758
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(2860..3051)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3860..4942)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=lacI
/protein_id="ANI69981.1"
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
gene complement(3860..4942)
/gene="lacI"
/label=lacI
promoter complement(4943..5020)
/gene="lacI"
/label=lacI promoter
promoter 5329..5347
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 5348..5372
/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)."
RBS 5387..5409
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 5426..5443
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
misc_feature 5450..6547
/label=similar to maltose binding protein
/note="similar to maltose binding protein"
CDS 5450..6547
/codon_start=1
/gene="malE (mutated)"
/product="maltose binding protein from E. coli"
/label=MBP
/note="This version of the gene does not encode a signal
sequence, so MBP will remain in the cytosol."
/translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
YAVRTAVINAASGRQTVDEALKDAQT"
CDS 6593..3
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
LOCUS Exported 6610 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pTEV19, complete sequence.
ACCESSION KU974159
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6610)
AUTHORS VanDrisse CM, Escalante-Semerena JC.
TITLE New high-cloning-efficiency vectors for complementation studies and
recombinant protein overproduction in Escherichia coli and
Salmonella enterica
JOURNAL Plasmid (2016) In press
PUBMED 27234933
REFERENCE 2 (bases 1 to 6610)
AUTHORS VanDrisse CM, Escalante-Semerena JC.
TITLE Direct Submission
JOURNAL Submitted (24-MAR-2016) Microbiology, University of Georgia, 120
Cedar St., Athens, GA 30602, USA
REFERENCE 3 (bases 1 to 6610)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6610
/organism="Expression vector pTEV19"
/mol_type="other DNA"
/db_xref="taxon:1856676"
CDS 56..73
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 140..187
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 224..679
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 706..810
/gene="bla"
/label=AmpR promoter
CDS 811..1671
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=bla
/protein_id="ANI69980.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene 811..1671
/gene="bla"
/label=bla
rep_origin 1842..2430
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 2616..2758
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(2860..3051)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3860..4942)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=lacI
/protein_id="ANI69981.1"
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
gene complement(3860..4942)
/gene="lacI"
/label=lacI
promoter complement(4943..5020)
/gene="lacI"
/label=lacI promoter
promoter 5329..5347
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 5348..5372
/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)."
RBS 5387..5409
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 5426..5443
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
misc_feature 5450..6547
/label=similar to maltose binding protein
/note="similar to maltose binding protein"
CDS 5450..6547
/codon_start=1
/gene="malE (mutated)"
/product="maltose binding protein from E. coli"
/label=MBP
/note="This version of the gene does not encode a signal
sequence, so MBP will remain in the cytosol."
/translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
YAVRTAVINAASGRQTVDEALKDAQT"
CDS 6593..3
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"