LOCUS Exported 6130 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Gateway vector pUGW14 DNA, complete sequence.
ACCESSION AB626671
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6130)
AUTHORS Tanaka Y, Nakagawa T.
TITLE Gateway Vectors for transient expression
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6130)
AUTHORS Tanaka Y, Nakagawa T.
TITLE Direct Submission
JOURNAL Submitted (19-APR-2011) Contact:Tsuyoshi Nakagawa Shimane
University, Center for Integrated Research in Science; 1060
Nishikawatsu, Matsue, Shimane 690-8504, Japan
REFERENCE 3 (bases 1 to 6130)
AUTHORS .
TITLE Direct Submission
COMMENT constructed using pUC119.
FEATURES Location/Qualifiers
source 1..6130
/organism="Gateway vector pUGW14"
/mol_type="other DNA"
/db_xref="taxon:1007965"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
regulatory 276..1109
/regulatory_class="promoter"
/note="35S promoter"
promoter 763..1108
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
misc_feature complement(1130..1254)
/label=attR1
/note="attR1"
protein_bind 1130..1254
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 1291..1321
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 1375..2034
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=cat
/protein_id="BAK19548.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
gene 1375..2034
/gene="cat"
/label=cat
CDS 2376..2681
/codon_start=1
/gene="ccdB"
/label=ccdB
/protein_id="BAK19549.1"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
gene 2376..2681
/gene="ccdB"
/label=ccdB
misc_feature 2722..2846
/label=attR2
/note="attR2"
protein_bind complement(2722..2846)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
misc_feature 2857..2976
/label=3xHA
/note="3xHA"
CDS 2875..2964
/codon_start=1
/product="three tandem HA epitope tags"
/label=3xHA
/translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
regulatory 2982..3258
/regulatory_class="terminator"
/note="NOS terminator"
terminator 2998..3250
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(3259..3275)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 3488..3943
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4225..4329
/gene="bla"
/label=AmpR promoter
CDS 4330..5190
/codon_start=1
/gene="bla"
/product="beta lactamase"
/label=bla
/protein_id="BAK19550.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene 4330..5190
/gene="bla"
/label=bla
rep_origin complement(5351..5951)
/direction=LEFT
/label=ColE1 Ori
/note="ColE1 Ori"
rep_origin 5361..5949
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
LOCUS Exported 6130 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Gateway vector pUGW14 DNA, complete sequence.
ACCESSION AB626671
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6130)
AUTHORS Tanaka Y, Nakagawa T.
TITLE Gateway Vectors for transient expression
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6130)
AUTHORS Tanaka Y, Nakagawa T.
TITLE Direct Submission
JOURNAL Submitted (19-APR-2011) Contact:Tsuyoshi Nakagawa Shimane
University, Center for Integrated Research in Science; 1060
Nishikawatsu, Matsue, Shimane 690-8504, Japan
REFERENCE 3 (bases 1 to 6130)
AUTHORS .
TITLE Direct Submission
COMMENT constructed using pUC119.
FEATURES Location/Qualifiers
source 1..6130
/organism="Gateway vector pUGW14"
/mol_type="other DNA"
/db_xref="taxon:1007965"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
regulatory 276..1109
/regulatory_class="promoter"
/note="35S promoter"
promoter 763..1108
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
misc_feature complement(1130..1254)
/label=attR1
/note="attR1"
protein_bind 1130..1254
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 1291..1321
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 1375..2034
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=cat
/protein_id="BAK19548.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
gene 1375..2034
/gene="cat"
/label=cat
CDS 2376..2681
/codon_start=1
/gene="ccdB"
/label=ccdB
/protein_id="BAK19549.1"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
gene 2376..2681
/gene="ccdB"
/label=ccdB
misc_feature 2722..2846
/label=attR2
/note="attR2"
protein_bind complement(2722..2846)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
misc_feature 2857..2976
/label=3xHA
/note="3xHA"
CDS 2875..2964
/codon_start=1
/product="three tandem HA epitope tags"
/label=3xHA
/translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
regulatory 2982..3258
/regulatory_class="terminator"
/note="NOS terminator"
terminator 2998..3250
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(3259..3275)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 3488..3943
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4225..4329
/gene="bla"
/label=AmpR promoter
CDS 4330..5190
/codon_start=1
/gene="bla"
/product="beta lactamase"
/label=bla
/protein_id="BAK19550.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene 4330..5190
/gene="bla"
/label=bla
rep_origin complement(5351..5951)
/direction=LEFT
/label=ColE1 Ori
/note="ColE1 Ori"
rep_origin 5361..5949
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"