LOCUS       Exported                8775 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMK33-NTAP, complete sequence.
ACCESSION   AY727853
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8775)
  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 8775)
  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 8775)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8775
                     /organism="Cloning vector pMK33-NTAP"
                     /mol_type="other DNA"
                     /db_xref="taxon:298282"
     misc_feature    1..8775
                     /note="pMK33-NTAP and pMK33-CTAP vectors are designed for 
                     making protein fusions tagged with the TAP tag and for 
                     generating stable Drosophila cell lines under Hygromycin 
                     selection. TAP fusions are inducible with copper sulfate. 
                     The parental vector, pMK33 (pMt/Hy), was created by Michael
                     Koelle."
     misc_feature    1..8
                     /label=NotI site
                     /note="NotI site"
     misc_feature    9..2689
                     /note="pAT153 vector backbone; similar to pAT153 cloning 
                     vector in GenBank Accession Number L08853"
     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=copia LTR
                     /note="copia LTR"
     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="AAV33420.1"
                     /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..7534
                     /codon_start=1
                     /product="NTAP-tag"
                     /label=NTAP-tag
                     /protein_id="AAV33419.1"
                     /translation="MAGLAQHDEAVDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLK
                     DDPSQSANLLAEAKKLNDAQAPKVDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKD
                     DPSQSANLLAEAKKLNDAQAPKVDANCDIPTTASENLYFQGELKRRWKKNFIAVSAANR
                     FKKISSSGALDDDDK"
     CDS             7031..7204
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             7208..7378
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     CDS             7415..7435
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             7442..7519
                     /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             7520..7534
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     misc_feature    7535..7558
                     /note="polylinker; unique cloning sites for XhoI, BamHI, 
                     EcoRV, SpeI"