LOCUS       Exported                8577 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pCeMM-NTAP(GS)-Gw, complete sequence.
ACCESSION   EF143813
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8577)
  AUTHORS   Burckstummer T, Bennett KL, Preradovic A, Schutze G, Hantschel O, 
            Superti-Furga G, Bauch A.
  TITLE     An efficient tandem affinity purification procedure for interaction 
            proteomics in mammalian cells
  JOURNAL   Nat. Methods 3 (12), 1013-1019 (2006)
  PUBMED    17060908
REFERENCE   2  (bases 1 to 8577)
  AUTHORS   Burckstummer T, Bennett KL, Preradovic A, Schutze G, Hantschel O, 
            Bauch A, Superti-Furga G.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-NOV-2006) Directors Laboratory, Research Center for 
            Molecular Medicine, Lazarettgasse 19/3, Vienna 1090, Austria
REFERENCE   3  (bases 1 to 8577)
  AUTHORS   Burckstummer T, Bennett KL, Preradovic A, Schutze G, Hantschel O, 
            Bauch A, Superti-Furga G.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-DEC-2006) Directors Laboratory, Research Center for 
            Molecular Medicine, Lazarettgasse 19/3, Vienna 1090, Austria
REFERENCE   4  (bases 1 to 8577)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     On Dec 14, 2006 this sequence version replaced EF143813.1.
FEATURES             Location/Qualifiers
     source          1..8577
                     /organism="Expression vector pCeMM-NTAP(GS)-Gw"
                     /mol_type="other DNA"
                     /db_xref="taxon:415353"
     misc_feature    17..604
                     /label=IRES
                     /note="IRES"
     misc_feature    20..605
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             606..1325
                     /codon_start=1
                     /gene="GFP"
                     /product="GFP"
                     /label=GFP
                     /protein_id="ABL60857.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     gene            606..1325
                     /gene="GFP"
                     /label=GFP
     repeat_region   1491..2084
                     /label=3 LTR
                     /note="3 LTR"
     LTR             1613..2082
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             2167..3027
                     /codon_start=1
                     /product="ampicillin resistance"
                     /label=ampicillin resistance
                     /protein_id="ABL60858.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3030..3950
     rep_origin      3198..3786
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4074..4095
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4110..4140
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4148..4164
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4172..4188
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      4205..4781
                     /regulatory_class="promoter"
                     /note="hCMV promoter"
     enhancer        4270..4573
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4578..4781
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     repeat_region   4782..4967
                     /label=5 LTR
                     /note="5 LTR"
     misc_feature    5020..5377
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     misc_feature    5026..5835
                     /label=packaging signal
                     /note="packaging signal"
     CDS             5442..5858
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     regulatory      6205..6223
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     promoter        6205..6223
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6266..6793
                     /codon_start=1
                     /gene="NTAP(GS)"
                     /product="NTAP(GS)"
                     /label=NTAP(GS)
                     /protein_id="ABL60854.1"
                     /translation="MEFMGTPAVTTYKLVINGKTLKGETTTKAVDAETAEKAFKQYAND
                     NGVDGVWTYDDATKTFTVTEVNTPAVTTYKLVINGKTLKGETTTKAVDAETAEKAFKQY
                     ANDNGVDGVWTYDDATKTFTVTEIDENLYFQGGSMDEKTTGWRGGHVVEGLAGELEQLR
                     ARLEHHPQGQREP"
     gene            6266..6793
                     /gene="NTAP(GS)"
                     /label=NTAP(GS)
     CDS             6653..6673
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             6680..6793
                     /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_recomb     6869..6993
                     /label=attR1
                     /note="attR1"
     protein_bind    6869..6993
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        7018..7048
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             7102..7761
                     /codon_start=1
                     /product="chloramphenicol resistance"
                     /label=chloramphenicol resistance
                     /protein_id="ABL60855.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             8103..8408
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB"
                     /label=ccdB
                     /protein_id="ABL60856.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            8103..8408
                     /gene="ccdB"
                     /label=ccdB
     misc_recomb     8449..8573
                     /label=attR2
                     /note="attR2"
     protein_bind    complement(8449..8573)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"