Bischof J, Bjorklund M, Furger E, Schertel C, Taipale J, Basler K.
pEGFP-2.attB 载体载体图谱
pEGFP-2.attB 载体载体序列
LOCUS Exported 8865 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pEGFP-2.attB, complete sequence.
ACCESSION KC896843
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8865)
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 140 (11), 2434-2442 (2013)
PUBMED 23637332
REFERENCE 2 (bases 1 to 8865)
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 8865)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..8865
/organism="Cloning vector pEGFP-2.attB"
/mol_type="other DNA"
/db_xref="taxon:1331594"
misc_feature 1..4136
/label=white dominant eye marker
/note="white dominant eye marker"
CDS join(479..550,952..1225,1300..1954,2016..2331,2552..2683,
2754..3368)
/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"
misc_feature 4143..4176
/label=loxP site
/note="loxP site"
protein_bind complement(4143..4176)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 4189..4258
/label=multiple cloning site
/note="multiple cloning site"
CDS 4264..4983
/codon_start=1
/product="EGFP"
/label=EGFP
/note="EGFP reporter"
/protein_id="AGL42324.1"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
misc_feature 4996..5695
/label=SV40 polyA region
/note="SV40 polyA region"
intron 5073..5138
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 5268..5288
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 5560..5694
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
misc_recomb 5702..5986
/label=attB site
/note="attB site"
protein_bind 5723..5792
/label=attB
/bound_moiety="phage phi-C31 integrase"
/note="attB site for the phi-C31 integrase (Groth et al.,
2000)"
promoter complement(6011..6029)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(6050..6066)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 6074..6090
/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(6098..6128)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6143..6164
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6452..7040)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7211..8071)
/codon_start=1
/product="ampicillin resistance protein"
/label=ampicillin resistance protein
/protein_id="AGL42325.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(8072..8176)
/gene="bla"
/label=AmpR promoter
rep_origin complement(8202..8657)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 8799..8815
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 8822..8840
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
Bischof J, Bjorklund M, Furger E, Schertel C, Taipale J, Basler K.
pEGFP-2.attB 载体载体图谱
pEGFP-2.attB 载体载体序列
LOCUS Exported 8865 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pEGFP-2.attB, complete sequence.
ACCESSION KC896843
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8865)
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 140 (11), 2434-2442 (2013)
PUBMED 23637332
REFERENCE 2 (bases 1 to 8865)
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 8865)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..8865
/organism="Cloning vector pEGFP-2.attB"
/mol_type="other DNA"
/db_xref="taxon:1331594"
misc_feature 1..4136
/label=white dominant eye marker
/note="white dominant eye marker"
CDS join(479..550,952..1225,1300..1954,2016..2331,2552..2683,
2754..3368)
/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"
misc_feature 4143..4176
/label=loxP site
/note="loxP site"
protein_bind complement(4143..4176)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 4189..4258
/label=multiple cloning site
/note="multiple cloning site"
CDS 4264..4983
/codon_start=1
/product="EGFP"
/label=EGFP
/note="EGFP reporter"
/protein_id="AGL42324.1"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
misc_feature 4996..5695
/label=SV40 polyA region
/note="SV40 polyA region"
intron 5073..5138
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 5268..5288
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 5560..5694
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
misc_recomb 5702..5986
/label=attB site
/note="attB site"
protein_bind 5723..5792
/label=attB
/bound_moiety="phage phi-C31 integrase"
/note="attB site for the phi-C31 integrase (Groth et al.,
2000)"
promoter complement(6011..6029)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(6050..6066)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 6074..6090
/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(6098..6128)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6143..6164
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6452..7040)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7211..8071)
/codon_start=1
/product="ampicillin resistance protein"
/label=ampicillin resistance protein
/protein_id="AGL42325.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(8072..8176)
/gene="bla"
/label=AmpR promoter
rep_origin complement(8202..8657)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 8799..8815
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 8822..8840
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"