LOCUS       sequence-180125-         846 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Twinstrep-SUMO-huLwCas13a for recombinant protein bacterial 
            expression. Insert is human codon optimized but expresses well in 
            bacteria..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 846)
  AUTHORS   Gootenberg JS, Abudayyeh OO, Lee JW, Essletzbichler P, Dy AJ, Joung 
            J, Verdine V, Donghia N, Daringer NM, Freije CA, Myhrvold C, 
            Bhattacharyya RP, Livny J, Regev A, Koonin EV, Hung DT, Sabeti PC, 
            Collins JJ, Zhang F
  TITLE     Nucleic acid detection with CRISPR-Cas13a/C2c2.
  JOURNAL   Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321.
  PUBMED    28408723
REFERENCE   2  (bases 1 to 846)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science. 
            2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321."
COMMENT     Sequence Label: pC013 - Twinstrep-SUMO-huLwCas13a
FEATURES             Location/Qualifiers
     source          1..846
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(1..77)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     283..302
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     386..405
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        386..404
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    405..429
                     /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)."
     RBS             444..466
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             485..502
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             512..529
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             join(542..565,566..601,602..625)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="WSHPQFEKGGGSGGGSGGSAWSHPQFEK"
ORIGIN
        1 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
       61 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
      121 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa
      181 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
      241 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
      301 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
      361 gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat
      421 aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata ccatgggcag
      481 cagccatcat catcatcatc acagcagcgg cctggtgccg cgcggcagcc atatggctag
      541 ctggagccat ccgcagtttg aaaaaggtgg tggtagcggt ggtggttcag gtggtagtgc
      601 atggtcacac cctcagtttg agaaaatgtc ggactcagaa gtcaatcaag aagctaagcc
      661 agaggtcaag ccagaagtca agcctgagac tcacatcaat ttaaaggtgt ccgatggatc
      721 ttcagagatc ttcttcaaga tcaaaaagac cactccttta agaaggctga tggaagcgtt
      781 cgctaaaaga cagggtaagg aaatggactc cttaagattc ttgtacgacg gtattagaat
      841 ccaagc
//
