载体基本信息 | |||
载体名称 | pN-PURO-PTP | 载体抗性 | Ampicillin |
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载体长度 | 5650 bp | 载体类型 | Transfection vector |
载体来源 | Schimanski B, Nguyen TN, Gunzl A. |
pN-PURO-PTP 载体载体图谱
pN-PURO-PTP 载体载体序列
LOCUS Exported 5650 bp ds-DNA circular SYN 18-DEC-2018 DEFINITION Transfection vector pN-PURO-PTP, complete sequence. ACCESSION DQ172901 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5650) AUTHORS Schimanski B, Nguyen TN, Gunzl A. TITLE Highly efficient tandem affinity purification of trypanosome protein complexes based on a novel epitope combination JOURNAL Eukaryotic Cell 4 (11), 1942-1950 (2005) PUBMED 16278461 REFERENCE 2 (bases 1 to 5650) AUTHORS Schimanski B, Nguyen TN, Gunzl A. TITLE Direct Submission JOURNAL Submitted (17-AUG-2005) Department of Genetics and Developmental Biology, University of Connecticut Health Center, 263 Farmington Ave, Farmington, CT 06030, USA REFERENCE 3 (bases 1 to 5650) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..5650 /organism="Transfection vector pN-PURO-PTP" /mol_type="other DNA" /db_xref="taxon:343988" promoter 21..125 /gene="bla" /label=AmpR promoter CDS 126..986 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 1157..1745 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 2033..2054 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2069..2099 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2107..2123 /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 2131..2147 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 2168..2186 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" misc_feature 2199..2204 /note="unique SacI restriction site for cloning the resistance marker cassette" misc_feature 2204..2702 /label=HSP70 genes 2 and 3 intergenic region /note="HSP70 genes 2 and 3 intergenic region" CDS 2703..3302 /codon_start=1 /product="puromycin N-acetyltransferase" /label=puromycin N-acetyltransferase /protein_id="ABA01557.1" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" misc_feature 3315..4055 /label=beta-alpha tubulin intergenic region /note="beta-alpha tubulin intergenic region" misc_feature 4056..4061 /note="unique HindIII restriction site for cloning the resistance marker cassette and the PTP cassette" misc_feature 4496..4972 /label=PTP tag /note="PTP tag" CDS 4526..4699 /codon_start=1 /product="IgG-binding unit of Staphylococcus aureus protein A" /label=ProtA /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL AEAKKLNDAQAPK" CDS 4703..4873 /codon_start=1 /product="IgG-binding unit of Staphylococcus aureus protein A" /label=ProtA /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA EAKKLNDAQAPK" CDS 4910..4930 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" misc_feature 4973..4998 /note="multiple cloning site containing unique restriction sites for KpnI, ApaI, SnaBI and NotI for fusing the target protein to the PTP sequence" promoter complement(5007..5025) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5032..5048) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 5190..5645 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
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