LOCUS       Exported                9626 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUAST-NTAP(GS), complete sequence.
ACCESSION   EU663572
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9626)
  AUTHORS   Kyriakakis P, Tipping M, Abed L, Veraksa A.
  TITLE     Tandem affinity purification in Drosophila: The advantages of the 
            GS-TAP system
  JOURNAL   Fly (Austin) 2 (4) (2008) In press
  PUBMED    18719405
REFERENCE   2  (bases 1 to 9626)
  AUTHORS   Veraksa A, Kyriakakis P, Tipping M, Abed L.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-APR-2008) Biology Department, University of 
            Massachusetts Boston, 100 Morrissey Blvd., Boston, MA 02125, USA
REFERENCE   3  (bases 1 to 9626)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9626
                     /organism="Cloning vector pUAST-NTAP(GS)"
                     /mol_type="other DNA"
                     /note="mammalian GS-TAP vectors were originally described 
                     in the following publication: Burckstummer T. et al. (2006)
                     Nature Methods 3, 1013-1019; designed for protein fusions 
                     with the GS-TAP tag and inducible expression in transgenic 
                     Drosophila; orientation is reversed relative to the 
                     compiled sequence of pUAST in FlyBase"
                     /db_xref="taxon:546946"
     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..3976
                     /codon_start=1
                     /product="NTAP(GS) tag"
                     /label=NTAP
                     /protein_id="ACF35722.1"
                     /translation="MGTPAVTTYKLVINGKTLKGETTTKAVDAETAEKAFKQYANDNGV
                     DGVWTYDDATKTFTVTEVNTPAVTTYKLVINGKTLKGETTTKAVDAETAEKAFKQYAND
                     NGVDGVWTYDDATKTFTVTEIGENLYFQGIDENLYFQGGSMDEKTTGWRGGHVVEGLAG
                     ELEQLRARLEHHPQGQREP"
     CDS             3809..3829
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3836..3856
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3863..3976
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     misc_feature    3979..4024
                     /note="polylinker; unique cloning sites for EcoRI, 
                     EagI/NotI, XhoI, KpnI, XbaI"
     intron          4102..4167
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4297..4317
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4742..4876
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             join(5381..5452,5847..6120,6195..6849,6911..7226,7447..7578,
                     7649..8263)
                     /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    9041..9626
                     /label=P element 5 end