LOCUS       Exported                8003 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDV-NTAP-CYFP, complete sequence.
ACCESSION   EF028675
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8003)
  AUTHORS   Meima ME, Weening KE, Schaap P.
  TITLE     Vectors for expression of proteins with single or combinatorial 
            fluorescent protein and tandem affinity purification tags in 
            Dictyostelium
  JOURNAL   Protein Expr. Purif. 53 (2), 283-288 (2007)
  PUBMED    17296313
REFERENCE   2  (bases 1 to 8003)
  AUTHORS   Meima ME, Weening KE, Schaap P.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-SEP-2006) School of Life Sciences, University of 
            Dundee, Dow Street, Dundee DD1 5EH, United Kingdom
REFERENCE   3  (bases 1 to 8003)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8003
                     /organism="Cloning vector pDV-NTAP-CYFP"
                     /lab_host="Dictyostelium discoideum"
                     /mol_type="other DNA"
                     /note="Cloning vector pDV-NTAP-CYFP is used to fuse target 
                     proteins at the N-terminus to the TAP-tag and at the 
                     C-terminus to enhanced yellow fluorescent protein (EYFP)"
                     /db_xref="taxon:409545"
     misc_feature    12..554
                     /note="N-terminal TAP-tag; contains enterokinase 
                     phosphorylation site"
     CDS             48..221
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             225..395
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     CDS             432..452
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             459..536
                     /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             537..551
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     misc_feature    555..572
                     /label=multipe cloning site
                     /note="multipe cloning site"
     misc_feature    573..1286
                     /note="3 end of enhanced yellow fluorescent protein 
                     (EYFP)"
     CDS             573..1286
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     regulatory      1313..2293
                     /regulatory_class="terminator"
                     /note="2H3 terminator"
     CDS             3026..3856
                     /codon_start=1
                     /product="neomycin phosphotransferase"
                     /label=neomycin phosphotransferase
                     /protein_id="ABK40491.1"
                     /translation="MDGEDVQAGSFRMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSD
                     AAVFRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWL
                     LLGEVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAG
                     LVDQDDLDEEHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCG
                     RLGVADRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3062..3856
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        4746..4850
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4851..5711
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="ABK40490.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5882..6470
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      7695..8003
                     /regulatory_class="promoter"
                     /note="actin15 promoter"