Bischof J, Bjorklund M, Furger E, Schertel C, Taipale J, Basler K.
pUASg.attB 载体载体图谱
pUASg.attB 载体载体序列
LOCUS Exported 10293 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pUASg.attB, complete sequence.
ACCESSION KC896836
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10293)
AUTHORS Bischof J, Bjorklund M, Furger E, Schertel C, Taipale J, Basler K.
TITLE A versatile platform for creating a comprehensive UAS-ORFeome
library in Drosophila
JOURNAL Development (2013) In press
PUBMED 23637332
REFERENCE 2 (bases 1 to 10293)
AUTHORS Bischof J, Bjorklund M, Furger E, Schertel C, Taipale J, Basler K.
TITLE Direct Submission
JOURNAL Submitted (13-APR-2013) Institute of Molecular Life Sciences,
University of Zurich, Winterthurerstrasse 190, Zurich 8057,
Switzerland
REFERENCE 3 (bases 1 to 10293)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..10293
/organism="Cloning vector pUASg.attB"
/mol_type="other DNA"
/db_xref="taxon:1331598"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 600..616
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 623..641
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 667..4802
/label=white dominant eye marker
/note="white dominant eye marker"
CDS join(1145..1216,1618..1891,1966..2620,2682..2997,3218..3349,
3420..4034)
/codon_start=1
/gene="white"
/product="Drosophila white gene eye color pigment"
/label=mini-white
/note="This ia a modified version of the white gene lacking
part of the first intron."
/translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
regulatory 4842..5197
/regulatory_class="promoter"
/note="5xUAS-hsp70 promoter region"
protein_bind 4842..4936
/label=5X UAS
/bound_moiety="GAL4"
/note="five tandem copies of the ""ScaI site"" 17-mer
CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS)
that efficiently binds yeast Gal4 (Webster et al., 1988;
Pfeiffer et al., 2010)"
promoter 4955..5193
/label=hsp70 promoter
/note="Drosophila melanogaster hsp70Bb promoter"
misc_feature 5236..6939
/label=attR1-CmR-ccdB-attR2 Gateway cassette
/note="attR1-CmR-ccdB-attR2 Gateway cassette"
protein_bind 5236..5360
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 5385..5415
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 5469..6149
/codon_start=1
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
DPAY"
CDS 6469..6774
/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
VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(6815..6939)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 6953..7765
/regulatory_class="terminator"
/note="tubulin alpha 1 terminator"
misc_recomb 7796..8080
/label=attB site
/note="attB site"
protein_bind 7817..7886
/label=attB
/bound_moiety="phage phi-C31 integrase"
/note="attB site for the phi-C31 integrase (Groth et al.,
2000)"
promoter complement(8105..8123)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(8144..8160)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 8168..8184
/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)."
promoter complement(8192..8222)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8237..8258
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(8546..9134)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9305..10165)
/codon_start=1
/product="ampicillin resistance protein"
/label=ampicillin resistance protein
/protein_id="AGL42314.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(10166..10270)
/gene="bla"
/label=AmpR promoter
Bischof J, Bjorklund M, Furger E, Schertel C, Taipale J, Basler K.
pUASg.attB 载体载体图谱
pUASg.attB 载体载体序列
LOCUS Exported 10293 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pUASg.attB, complete sequence.
ACCESSION KC896836
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10293)
AUTHORS Bischof J, Bjorklund M, Furger E, Schertel C, Taipale J, Basler K.
TITLE A versatile platform for creating a comprehensive UAS-ORFeome
library in Drosophila
JOURNAL Development (2013) In press
PUBMED 23637332
REFERENCE 2 (bases 1 to 10293)
AUTHORS Bischof J, Bjorklund M, Furger E, Schertel C, Taipale J, Basler K.
TITLE Direct Submission
JOURNAL Submitted (13-APR-2013) Institute of Molecular Life Sciences,
University of Zurich, Winterthurerstrasse 190, Zurich 8057,
Switzerland
REFERENCE 3 (bases 1 to 10293)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..10293
/organism="Cloning vector pUASg.attB"
/mol_type="other DNA"
/db_xref="taxon:1331598"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 600..616
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 623..641
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 667..4802
/label=white dominant eye marker
/note="white dominant eye marker"
CDS join(1145..1216,1618..1891,1966..2620,2682..2997,3218..3349,
3420..4034)
/codon_start=1
/gene="white"
/product="Drosophila white gene eye color pigment"
/label=mini-white
/note="This ia a modified version of the white gene lacking
part of the first intron."
/translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
regulatory 4842..5197
/regulatory_class="promoter"
/note="5xUAS-hsp70 promoter region"
protein_bind 4842..4936
/label=5X UAS
/bound_moiety="GAL4"
/note="five tandem copies of the ""ScaI site"" 17-mer
CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS)
that efficiently binds yeast Gal4 (Webster et al., 1988;
Pfeiffer et al., 2010)"
promoter 4955..5193
/label=hsp70 promoter
/note="Drosophila melanogaster hsp70Bb promoter"
misc_feature 5236..6939
/label=attR1-CmR-ccdB-attR2 Gateway cassette
/note="attR1-CmR-ccdB-attR2 Gateway cassette"
protein_bind 5236..5360
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 5385..5415
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 5469..6149
/codon_start=1
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
DPAY"
CDS 6469..6774
/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
VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(6815..6939)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 6953..7765
/regulatory_class="terminator"
/note="tubulin alpha 1 terminator"
misc_recomb 7796..8080
/label=attB site
/note="attB site"
protein_bind 7817..7886
/label=attB
/bound_moiety="phage phi-C31 integrase"
/note="attB site for the phi-C31 integrase (Groth et al.,
2000)"
promoter complement(8105..8123)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(8144..8160)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 8168..8184
/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)."
promoter complement(8192..8222)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8237..8258
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(8546..9134)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9305..10165)
/codon_start=1
/product="ampicillin resistance protein"
/label=ampicillin resistance protein
/protein_id="AGL42314.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(10166..10270)
/gene="bla"
/label=AmpR promoter