LOCUS       Exported                8793 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMK33-NTAP(GS), complete sequence.
ACCESSION   EU663570
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8793)
  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 8793)
  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 8793)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8793
                     /organism="Cloning vector pMK33-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 making protein 
                     fusions tagged with the GS-TAP tag and for generating 
                     stable Drosophila cell lines under hygromycin selection; 
                     GS-TAP fusions are inducible with copper sulfate"
                     /db_xref="taxon:546944"
     misc_feature    9..2689
                     /note="pAT153 vector backbone; Amp resistance; low copy 
                     number"
     rep_origin      complement(865..1453)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1624..2484)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2485..2589)
                     /gene="bla"
                     /label=AmpR promoter
     repeat_region   4070..4354
                     /label=derived from copia transposon
                     /note="derived from copia transposon"
     promoter        4071..4350
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             4396..5421
                     /codon_start=1
                     /product="hygromycin phosphotransferase"
                     /label=hygromycin phosphotransferase
                     /protein_id="ACF35718.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     intron          5784..5849
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5979..5999
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6424..6558
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      6559..7000
                     /regulatory_class="promoter"
                     /note="metallothionein promoter"
     promoter        6559..6985
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     CDS             7001..7546
                     /codon_start=1
                     /product="NTAP(GS) tag"
                     /label=NTAP
                     /protein_id="ACF35719.1"
                     /translation="MGTPAVTTYKLVINGKTLKGETTTKAVDAETAEKAFKQYANDNGV
                     DGVWTYDDATKTFTVTEVNTPAVTTYKLVINGKTLKGETTTKAVDAETAEKAFKQYAND
                     NGVDGVWTYDDATKTFTVTEIGENLYFQGIDENLYFQGGSMDEKTTGWRGGHVVEGLAG
                     ELEQLRARLEHHPQGQREP"
     CDS             7379..7399
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             7406..7426
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             7433..7546
                     /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    7547..7576
                     /note="unique cloning sites: XhoI, BamHI, EcoRV, KpnI, 
                     SpeI; polylinker"