LOCUS       Exported                9528 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  dPspCas13b-ADAR2DD(E488Q/T375G) fusion that can be used to 
            selectively edit adenosine to inosine in RNA when used in conjuction
            with a guide RNA. Cas13b is truncated to allow for expression by 
            AAV.
ACCESSION   .
VERSION     .
KEYWORDS    pC0055-CMV-dPspCas13b-GS-ADAR2DD(E488Q/T375G)-delta-984-1090
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9528)
  AUTHORS   Cox DBT, Gootenberg JS, Abudayyeh OO, Franklin B, Kellner MJ, Joung 
            J, Zhang F
  TITLE     RNA editing with CRISPR-Cas13.
  JOURNAL   Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180.
  PUBMED    29070703
REFERENCE   2  (bases 1 to 9528)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science. 
            2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180."
FEATURES             Location/Qualifiers
     source          1..9528
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1241..1267
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(2476..2493)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2482..2706
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2752..3180
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2839..2858)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3049..3070
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3189..3209)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3194..3523
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3374..3509
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3436..3455
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3590..4384
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3644..3663)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4254..4273
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4558..4679
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4595..4614)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4649..4668
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4728..4744)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4728..4744)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4741..4763)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4752..4768
                     /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)."
     promoter        complement(4776..4806)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4821..4842
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4959..4976)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5130..5718)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5210..5229)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5889..6749)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6512..6531
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6750..6854)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6929..6948)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7120..7499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7500..7703
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7654..7674
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7748..7767
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7748..7766
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"