下载GeneBank文件(.gb)

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"