AGG1521-pCHIKVRep1 vector (V015673)

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LOCUS       62056_316        5623 bp DNA     circular SYN 12-AUG-2020
DEFINITION  Cloning vector AGG1521:pCHIKVRep1, complete sequence.
ACCESSION   MT119957
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5623)
  AUTHORS   Tng PYL., Carabajal Paladino L, Verkuijl SAN., Purcell J, Merits A, 
            Leftwich PT, Fragkoudis R, Noad R, Alphey L.
  TITLE     Cas13b-dependent and Cas13b-independent RNA knockdown of viral 
            sequences in mosquito cells following guide RNA expression
  JOURNAL   Commun Biol 3 (1), 413 (2020)
  PUBMED    32737398
REFERENCE   2  (bases 1 to 5623)
  AUTHORS   Tng PYL., Carabajal Paladino L, Verkuijl S, Purcell J, Merits A, 
            Leftwich PT, Fragkoudis R, Noad R, Alphey L.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-FEB-2020) Arthropod Genetics Group, The Pirbright 
            Institute, Ash Road, Pirbright, Woking, Surrey GU24 0NF, UK
REFERENCE   3  (bases 1 to 5623)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol"; date: "2020"; volume: "3"; issue: "1"; pages: "413"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-FEB-2020) Arthropod Genetics Group, The Pirbright Institute, Ash
            Road, Pirbright, Woking, Surrey GU24 0NF, UK"
FEATURES             Location/Qualifiers
     source          1..5623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(227..1033)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1525..1541
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2526..3242
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             3424..3936
                     /codon_start=1
                     /label=Nluc
                     /note="NanoLuc(R) luciferase"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     polyA_signal    4533..4654
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4679..4695)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4703..4719)
                     /label=lac operator
                     /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)."
     promoter        complement(4727..4757)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4772..4793)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(join(5081..5623,1..46))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"