LOCUS       Exported                9620 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUAST-NTAP, complete sequence.
ACCESSION   AY727855
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9620)
  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 9620)
  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 9620)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9620
                     /organism="Cloning vector pUAST-NTAP"
                     /mol_type="other DNA"
                     /db_xref="taxon:298284"
     misc_feature    1..9620
                     /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"
     CDS             3431..3970
                     /codon_start=1
                     /product="NTAP-tag"
                     /label=NTAP-tag
                     /protein_id="AAV33423.1"
                     /translation="MAGLAQHDEAVDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLK
                     DDPSQSANLLAEAKKLNDAQAPKVDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKD
                     DPSQSANLLAEAKKLNDAQAPKVDANCDIPTTASENLYFQGELKRRWKKNFIAVSAANR
                     FKKISSSGALDDDDKLI"
     CDS             3461..3634
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             3638..3808
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     CDS             3845..3865
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3872..3949
                     /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             3950..3964
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     misc_feature    3973..4018
                     /note="polylinker; unique cloning sites for EcoRI, 
                     EagI/NotI, XhoI, KpnI, XbaI"
     intron          4096..4161
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4291..4311
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4736..4870
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             join(5375..5446,5841..6114,6189..6843,6905..7220,7441..7572,
                     7643..8257)
                     /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    9035..9620
                     /label=P element 5 end