LOCUS Exported 6669 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Gateway vector pUGW55 DNA, complete sequence.
ACCESSION AB626690
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6669)
AUTHORS Tanaka Y, Nakagawa T.
TITLE Gateway Vectors for transient expression
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6669)
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 6669)
AUTHORS .
TITLE Direct Submission
COMMENT constructed using pUC119.
FEATURES Location/Qualifiers
source 1..6669
/organism="Gateway vector pUGW55"
/mol_type="other DNA"
/db_xref="taxon:1007984"
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 258..1091
/regulatory_class="promoter"
/note="35S promoter"
promoter 745..1090
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
misc_feature 1111..1785
/label=monomeric red fluorescent protein
/note="monomeric red fluorescent protein"
CDS 1111..1785
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/note="mammalian codon-optimized"
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
misc_feature complement(1795..1919)
/label=attR1
/note="attR1"
protein_bind 1795..1919
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 1956..1986
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 2040..2699
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=cat
/protein_id="BAK19609.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
gene 2040..2699
/gene="cat"
/label=cat
CDS 3041..3346
/codon_start=1
/gene="ccdB"
/label=ccdB
/protein_id="BAK19610.1"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
gene 3041..3346
/gene="ccdB"
/label=ccdB
misc_feature 3387..3511
/label=attR2
/note="attR2"
protein_bind complement(3387..3511)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 3521..3797
/regulatory_class="terminator"
/note="NOS terminator"
terminator 3537..3789
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(3798..3814)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 4027..4482
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4764..4868
/gene="bla"
/label=AmpR promoter
CDS 4869..5729
/codon_start=1
/gene="bla"
/product="beta lactamase"
/label=bla
/protein_id="BAK19611.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene 4869..5729
/gene="bla"
/label=bla
rep_origin complement(5890..6490)
/direction=LEFT
/label=ColE1 Ori
/note="ColE1 Ori"
rep_origin 5900..6488
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
LOCUS Exported 6669 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Gateway vector pUGW55 DNA, complete sequence.
ACCESSION AB626690
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6669)
AUTHORS Tanaka Y, Nakagawa T.
TITLE Gateway Vectors for transient expression
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6669)
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 6669)
AUTHORS .
TITLE Direct Submission
COMMENT constructed using pUC119.
FEATURES Location/Qualifiers
source 1..6669
/organism="Gateway vector pUGW55"
/mol_type="other DNA"
/db_xref="taxon:1007984"
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 258..1091
/regulatory_class="promoter"
/note="35S promoter"
promoter 745..1090
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
misc_feature 1111..1785
/label=monomeric red fluorescent protein
/note="monomeric red fluorescent protein"
CDS 1111..1785
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/note="mammalian codon-optimized"
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
misc_feature complement(1795..1919)
/label=attR1
/note="attR1"
protein_bind 1795..1919
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 1956..1986
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 2040..2699
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=cat
/protein_id="BAK19609.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
gene 2040..2699
/gene="cat"
/label=cat
CDS 3041..3346
/codon_start=1
/gene="ccdB"
/label=ccdB
/protein_id="BAK19610.1"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
gene 3041..3346
/gene="ccdB"
/label=ccdB
misc_feature 3387..3511
/label=attR2
/note="attR2"
protein_bind complement(3387..3511)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 3521..3797
/regulatory_class="terminator"
/note="NOS terminator"
terminator 3537..3789
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(3798..3814)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 4027..4482
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4764..4868
/gene="bla"
/label=AmpR promoter
CDS 4869..5729
/codon_start=1
/gene="bla"
/product="beta lactamase"
/label=bla
/protein_id="BAK19611.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene 4869..5729
/gene="bla"
/label=bla
rep_origin complement(5890..6490)
/direction=LEFT
/label=ColE1 Ori
/note="ColE1 Ori"
rep_origin 5900..6488
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"