AviTag-SpyCatcher vector (V000988)

Basic Vector Information

      • Vector Name:
      • AviTag-SpyCatcher
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 4115 bp
      • Type:
      • Bacterial Expression
      • Copy Number:
      • High Copy
      • Cloning Method:
      • Ligation Independent Cloning
      • 5' Primer:
      • TAA TAC GAC TCA CTA TAG GG

AviTag-SpyCatcher vector Vector Map

AviTag-SpyCatcher4115 bp60012001800240030003600T7RBS6xHisAviTag(TM)TEV site6xHisT7 Termf1 oriAmpR promoterAmpRoriL4440bompGEX 3'

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 ℃.

AviTag-SpyCatcher vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                4115 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Bacterial expression of SpyCatcher with a peptide tag enabling 
            site-specific biotinylation..
ACCESSION   .
VERSION     .
KEYWORDS    AviTag-SpyCatcher
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4115)
  AUTHORS   Veggiani G, Nakamura T, Brenner MD, Gayet RV, Yan J, Robinson CV, 
            Howarth M
  TITLE     Programmable polyproteams built using twin peptide superglues.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Jan 19. pii: 201519214.
  PUBMED    26787909
REFERENCE   2  (bases 1 to 4115)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A. 2016 Jan 19. pii: 201519214."
FEATURES             Location/Qualifiers
     source          1..4115
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     21..40
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             71..93
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             113..130
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             137..181
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             206..226
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             636..653
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(706..724)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      720..767
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      804..1259
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(891..910)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1101..1122
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1285..1389
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1390..2250
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1608..1627)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2421..3009
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2910..2929
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3163..3180
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3195..3334
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3260..3279)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3420..3442
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3436..3627)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"

This page is informational only.