pN-PURO-PTP 载体 (V004369#)

载体名称:
pN-PURO-PTP
载体抗性:
Ampicillin
载体长度:
5650 bp
载体类型:
Transfection vector
复制子:
ori
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 regionPuroRbeta-alpha tubulin intergenic regionunique HindIII restriction site for cloning the resistance marker cassette and the PTP cassetteProtAProtATEV sitemultiple 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       40924_32963        5650 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Transfection vector pN-PURO-PTP, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5650)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Eukaryotic 
            Cell"; date: "2005"; volume: "4"; issue: "11"; pages: "1942-1950"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (17-AUG-2005) Department of Genetics and Developmental Biology, 
            University of Connecticut Health Center, 263 Farmington Ave, 
            Farmington, CT 06030, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..125
                     /label=AmpR promoter
     CDS             126..983
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1157..1745
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2033..2054
                     /label=CAP binding site
                     /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
                     /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..3299
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     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"
     CDS             4526..4699
                     /label=ProtA
                     /note="IgG-binding unit of Staphylococcus aureus protein A"
     CDS             4703..4873
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     CDS             4910..4930
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     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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