NovoPro

pET His6-TwinStrep-SUMO-LwCas13a vector

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Cat No. Plasmid Name Status Add to cart
V10159 pET His6-TwinStrep-SUMO-LwCas13a

Basic Vector Information

LwCas13a refers to Cas13a from Leptotrichia wadei. CRISPR adaptive immunity pathways protect prokaryotic cells against foreign nucleic acids using CRISPR RNA (crRNA)-guided nucleases. In type VI-A CRISPR-Cas systems, the signature protein Cas13a (formerly C2c2) contains two separate ribonuclease activities that catalyze crRNA maturation and ssRNA degradation. The Cas13a protein family occurs across different bacterial phyla and varies widely in both protein sequence and corresponding crRNA sequence conservation. Although grouped phylogenetically together, the Cas13a enzyme family comprises two distinct functional groups that recognize orthogonal sets of crRNAs and possess different ssRNA cleavage specificities. These functional distinctions could not be bioinformatically predicted, suggesting more subtle co-evolution of Cas13a enzymes. Additionally, Cas13a pre-crRNA processing is not essential for ssRNA cleavage, although it enhances ssRNA targeting for crRNAs encoded internally within the CRISPR array. Download Sequencing Data for His6-TwinStrep-SUMO-LwCas13a(*.ab1)

Basic Vector Information
Vector Name pET His6-TwinStrep-SUMO-LwCas13a Antibiotic Resistance Ampicillin
Length 9549 bp Type pET & Duet Vectors (Novagen)
Source NovoPro Copy Number High copy number
Promoter T7 Cloning Method Enzyme Cut
5' Primer T7 promoter 3' Primer T7 terminator
Fusion Tag 6*His, Strep-Tag II, SUMO Expression Method IPTG induced

pET His6-TwinStrep-SUMO-LwCas13a vector Vector Map

pET His6-TwinStrep-SUMO-LwCas13a vector Vector Map

pET His6-TwinStrep-SUMO-LwCas13a vector Sequence

LOCUS       pET-His6-SUMO-LwCas13a        9549 bp    DNA     circular
DEFINITION  .
KEYWORDS    pET-His6-SUMO-LwCas13a.
SOURCE      synthetic DNA construct.
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9549)
  AUTHORS   xsan
  TITLE     Direct Submission
            http://www.invitrogen.com/
FEATURES             Location/Qualifiers
     source          1..9549
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /vntifkey="98"
     promoter        104..208
                     /gene="bla"
                     /vntifkey="30"
                     /label=bla
                     /note="AmpR promoter"
     CDS             209..1069
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /vntifkey="4"
                     /label=bla
                     /note="AmpR confers resistance to ampicillin, carbenicillin, and related antibiotics"
     rep_origin      1240..1828
                     /direction=RIGHT
                     /vntifkey="33"
                     /label=Rep_Origin_1
                     /note="ori high-copy-number ColE1/pMB1/pBR322/pUC origin of  replication"
     misc_feature    2014..2156
                     /vntifkey="21"
                     /label=Misc_Feature_1
                     /note="bom basis of mobility region from pBR322"
     CDS             complement(2258..2449)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /vntifkey="4"
                     /label=rop
                     /note="rop"
     CDS             complement(3761..4843)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /vntifkey="4"
                     /label=lacI
                     /note="lacI 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)."
     promoter        complement(4844..4921)
                     /gene="lacI"
                     /vntifkey="30"
                     /label=lacI
                     /note="lacI promoter"
     promoter        5230..5248
                     /vntifkey="30"
                     /label=Promoter_P_1
                     /note="T7 promoter promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5249..5273
                     /bound_moiety="lac repressor encoded by lacI"
                     /vntifkey="31"
                     /label=Protein_Bind_1
                     /note="lac operator 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)."
     CDS             5329..5346
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /vntifkey="4"
                     /label=CDS_1
                     /note="6xHis"
     CDS             5356..5373
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /vntifkey="4"
                     /label=CDS_2
                     /note="thrombin site"
     CDS             5386..5409
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /vntifkey="4"
                     /label=CDS_3
                     /note="Strep-Tag II"
     CDS             5446..5469
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /vntifkey="4"
                     /label=CDS_4
                     /note="Strep-Tag II"
     CDS             5470..5761
                     /codon_start=1
                     /vntifkey="4"
                     /label=CDS_5
                     /note="SUMO"
     CDS             5767..9222
                     /codon_start=1
                     /vntifkey="4"
                     /label=CDS_6
                     /note="LwCas13a"
     terminator      9290..9337
                     /vntifkey="43"
                     /label=Terminator_1
                     /note="T7 terminator transcription terminator for bacteriophage T7 RNA  polymerase"
     promoter        complement(9512..9540)
                     /vntifkey="30"
                     /label=Promoter_P_2
                     /note="tet promoter E. coli promoter for tetracycline efflux protein  gene"