pN-PURO-PTP 载体 (V004369#)

载体基本信息
载体名称 pN-PURO-PTP 载体抗性 Ampicillin
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-PTP5650 bp60012001800240030003600420048005400AmpR promoterAmpRoriCAP binding sitelac promoterlac operatorM13 revT3 promoterunique SacI restriction site for cloning the resistance marker cassetteHSP70 genes 2 and 3 intergenic regionpuromycin N-acetyltransferasebeta-alpha tubulin intergenic regionunique HindIII restriction site for cloning the resistance marker cassette and the PTP cassettePTP tagmultiple cloning site containing unique restriction sites for KpnI, ApaI, SnaBI and NotI for fusing the target protein to the PTP sequenceT7 promoterM13 fwdf1 ori

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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