R777-E011 Hs.ARHGAP35 vector (V001282)

Basic Vector Information

      • Vector Name:
      • R777-E011 Hs.ARHGAP35
      • Antibiotic Resistance:
      • Spectinomycin
      • Length:
      • 7193 bp
      • Type:
      • Gateway Entry Clone
      • Copy Number:
      • High Copy
      • Cloning Method:
      • Gateway Cloning
      • 5' Primer:
      • M13F 23-mer (5'-CCCAGTCACGACGTTGTAAAACG)
      • 3' Primer:
      • attLR 23-mer (5'-TAACATCAGAGATTTTGAGACAC)

R777-E011 Hs.ARHGAP35 vector Vector Map

R777-E011 Hs.ARHGAP357193 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900In lacZ geneattL1RBSattL2SmRoriL4440rrnB T2 terminatorrrnB T1 terminator

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

R777-E011 Hs.ARHGAP35 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                7193 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Gateway ORF clone of human ARHGAP35 [NM_004491.4] with stop codon 
            (for native or N-terminal fusions).
ACCESSION   .
VERSION     .
KEYWORDS    R777-E011 Hs.ARHGAP35
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7193)
  TITLE     Esposito lab
REFERENCE   2  (bases 1 to 7193)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7193
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     23..45
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     37..54
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     38..54
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    84..183
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     RBS             4256..4264
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     protein_bind    complement(4684..4783)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             5160..5951
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      6044..6632
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6533..6552
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6786..6803
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      6962..6989
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      7081..7167
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"

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