pN-PURO-PTP vector (V004369)

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"