LOCUS Exported 9627 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pUAST-CTAP, complete sequence.
ACCESSION AY727856
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9627)
AUTHORS Veraksa A, Bauer A, Artavanis-Tsakonas S.
TITLE Analyzing protein complexes in Drosophila with tandem affinity
purification-mass spectrometry
JOURNAL Dev. Dyn. 232 (3), 827-834 (2005)
PUBMED 15704125
REFERENCE 2 (bases 1 to 9627)
AUTHORS Veraksa A, Bauer A, Artavanis-Tsakonas S.
TITLE Direct Submission
JOURNAL Submitted (17-AUG-2004) MGH Cancer Center, Harvard Medical School,
Bldg 149, 13th Street, Charlestown, MA 02129, USA
REFERENCE 3 (bases 1 to 9627)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..9627
/organism="Cloning vector pUAST-CTAP"
/mol_type="other DNA"
/db_xref="taxon:298285"
misc_feature 1..9627
/note="pUAST-NTAP and pUAST-CTAP vectors are designed for
protein fusions with the TAP tag and inducible expression
in transgenic Drosophila. Note that the orientation of
these vectors is reversed relative to the compiled sequence
of pUAST in FlyBase."
rep_origin complement(235..823)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(994..1854)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(1855..1959)
/gene="bla"
/label=AmpR promoter
misc_feature 2761..2993
/label=P element 3 end
/note="P element 3 end"
protein_bind 3055..3149
/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 3168..3406
/label=hsp70 promoter
/note="Drosophila melanogaster hsp70Bb promoter"
misc_feature 3420..3465
/note="polylinker; unique cloning sites for EcoRI,
EagI/NotI, XhoI, KpnI, XbaI; XbaI site is dam methylation
sensitive"
CDS 3469..4020
/codon_start=1
/product="CTAP-tag"
/label=CTAP-tag
/protein_id="AAV33424.1"
/translation="MEKRRWKKNFIAVSAANRFKKISSSGALDYDIPTTASENLYFQGE
LKTAALAQHDEAVDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLAE
AKKLNDAQAPKVDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLAEA
KKLNGAQAPKVDANSAGKST"
CDS 3475..3552
/codon_start=1
/product="calmodulin-binding peptide"
/label=CBP
/note="derived from skeletal muscle myosin light chain
kinase; binds calmodulin with nanomolar affinity in the
presence of calcium"
/translation="KRRWKKNFIAVSAANRFKKISSSGAL"
CDS 3580..3600
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 3640..3813
/codon_start=1
/product="IgG-binding unit of Staphylococcus aureus protein
A"
/label=ProtA
/translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
AEAKKLNDAQAPK"
CDS 3817..3987
/codon_start=1
/product="IgG-binding unit of Staphylococcus aureus protein
A"
/label=ProtA
/translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
EAKKLNGAQAPK"
intron 4103..4168
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 4298..4318
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 4743..4877
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
CDS join(5382..5453,5848..6121,6196..6850,6912..7227,7448..7579,
7650..8264)
/codon_start=1
/gene="white"
/product="Drosophila white gene eye color pigment"
/label=mini-white
/note="This is a modified version of the white gene lacking
part of the first intron."
/translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
GTLLPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
misc_feature 9042..9627
/label=P element 5 end
LOCUS Exported 9627 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pUAST-CTAP, complete sequence.
ACCESSION AY727856
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9627)
AUTHORS Veraksa A, Bauer A, Artavanis-Tsakonas S.
TITLE Analyzing protein complexes in Drosophila with tandem affinity
purification-mass spectrometry
JOURNAL Dev. Dyn. 232 (3), 827-834 (2005)
PUBMED 15704125
REFERENCE 2 (bases 1 to 9627)
AUTHORS Veraksa A, Bauer A, Artavanis-Tsakonas S.
TITLE Direct Submission
JOURNAL Submitted (17-AUG-2004) MGH Cancer Center, Harvard Medical School,
Bldg 149, 13th Street, Charlestown, MA 02129, USA
REFERENCE 3 (bases 1 to 9627)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..9627
/organism="Cloning vector pUAST-CTAP"
/mol_type="other DNA"
/db_xref="taxon:298285"
misc_feature 1..9627
/note="pUAST-NTAP and pUAST-CTAP vectors are designed for
protein fusions with the TAP tag and inducible expression
in transgenic Drosophila. Note that the orientation of
these vectors is reversed relative to the compiled sequence
of pUAST in FlyBase."
rep_origin complement(235..823)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(994..1854)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(1855..1959)
/gene="bla"
/label=AmpR promoter
misc_feature 2761..2993
/label=P element 3 end
/note="P element 3 end"
protein_bind 3055..3149
/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 3168..3406
/label=hsp70 promoter
/note="Drosophila melanogaster hsp70Bb promoter"
misc_feature 3420..3465
/note="polylinker; unique cloning sites for EcoRI,
EagI/NotI, XhoI, KpnI, XbaI; XbaI site is dam methylation
sensitive"
CDS 3469..4020
/codon_start=1
/product="CTAP-tag"
/label=CTAP-tag
/protein_id="AAV33424.1"
/translation="MEKRRWKKNFIAVSAANRFKKISSSGALDYDIPTTASENLYFQGE
LKTAALAQHDEAVDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLAE
AKKLNDAQAPKVDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLAEA
KKLNGAQAPKVDANSAGKST"
CDS 3475..3552
/codon_start=1
/product="calmodulin-binding peptide"
/label=CBP
/note="derived from skeletal muscle myosin light chain
kinase; binds calmodulin with nanomolar affinity in the
presence of calcium"
/translation="KRRWKKNFIAVSAANRFKKISSSGAL"
CDS 3580..3600
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 3640..3813
/codon_start=1
/product="IgG-binding unit of Staphylococcus aureus protein
A"
/label=ProtA
/translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
AEAKKLNDAQAPK"
CDS 3817..3987
/codon_start=1
/product="IgG-binding unit of Staphylococcus aureus protein
A"
/label=ProtA
/translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
EAKKLNGAQAPK"
intron 4103..4168
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 4298..4318
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 4743..4877
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
CDS join(5382..5453,5848..6121,6196..6850,6912..7227,7448..7579,
7650..8264)
/codon_start=1
/gene="white"
/product="Drosophila white gene eye color pigment"
/label=mini-white
/note="This is a modified version of the white gene lacking
part of the first intron."
/translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
GTLLPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
misc_feature 9042..9627
/label=P element 5 end