LOCUS       sequence-361526-        5782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector carrying a guide RNA targeting the human FANCC mRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5782)
  AUTHORS   Katrekar D, Yen J, Xiang Y, Saha A, Meluzzi D, Savva Y, Mali P
  TITLE     Efficient in vitro and in vivo RNA editing via recruitment of 
            endogenous ADARs using circular guide RNAs.
  JOURNAL   Nat Biotechnol. 2022 Feb 10. pii: 10.1038/s41587-021-01171-4. doi: 
            10.1038/s41587-021-01171-4.
  PUBMED    35145312
REFERENCE   2  (bases 1 to 5782)
  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: "Nat 
            Biotechnol. 2022 Feb 10. pii: 10.1038/s41587-021-01171-4. doi: 
            10.1038/s41587-021-01171-4."
COMMENT     Sequence Label: Circular 200,100 with interspersed loops (FANCC)
FEATURES             Location/Qualifiers
     source          1..5782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        160..408
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     160..180
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     331..350
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        820..1133
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     1052..1072
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     protein_bind    complement(1145..1169)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     enhancer        1184..1563
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1564..1767
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1718..1738
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1764..1788
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1879..1888
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1885..1887,1888..1890,1891..2601)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1885..1887,1888..1890,1891..2601)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1930..1951)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2191..2210)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2538..2559
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2680..2699)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2724..2772
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     repeat_region   2987..3127
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2987..3116
                     /label=AAV2 ITR
     primer_bind     complement(3167..3183)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3167..3183)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3180..3202)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3191..3207
                     /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(join(3215..3221,3222..3239,3240..3245))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3260..3281
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3569..4157)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3649..3668)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4328..5143)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4328..5143)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5051..5070
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5144..5248)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5316..5334
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5372..5394)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5494..5513
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5707..5729
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5722..5738
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5763..5779
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtggagg agggcctatt tcccatgatt
      181 ccttcatatt tgcatatacg atacaaggct gttagagaga taattagaat taatttgact
      241 gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag
      301 tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag
      361 tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgc catcagtcgc
      421 cggtcccaag cccggataaa atgggagggg gcgggaaacc gcctaaccat gccgactgat
      481 ggcagaaaaa aaaaaagagt ctgggctgag ggacctggct ctgctaaatg taaaatagat
      541 acttcgtgat tgtcccaaga tgacatcagc tcattctcac agcccagcga gggcacctac
      601 tctataaatg cgtggccaca gcagttcacc tgtcctgtgg gggaggcgag cctgatccca
      661 ggtgccgggc acccacacgg cctgcgtggc gtctagaaaa aaaaaactgc catcagtcgg
      721 cgtggactgt agaacactgc caatgccggt cccaagcccg gataaaagtg gagggtacag
      781 tccacgcttt tttcgtactg agtcgcccag tctcagatag atccgacgcc gccatctcta
      841 ggcccgcgcc ggccccctcg cacagacttg tgggagaagc tcggctactc ccctgccccg
      901 gttaatttgc atataatatt tcctagtaac tatagaggct taatgtgcga taaaagacag
      961 ataatctgtt ctttttaata ctagctacat tttacatgat aggcttggat ttctataaga
     1021 gatacaaata ctaaattatt attttaaaaa acagcacaaa aggaaactca ccctaactgt
     1081 aaagtaattg tgtgttttga gactataaat atcccttgga gaaaagcctt gtttggattc
     1141 ctagacccag ctttcttgta caaagttggc attaatacgc gttgacattg attattgact
     1201 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     1261 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     1321 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     1381 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     1441 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     1501 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     1561 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     1621 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     1681 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     1741 cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc
     1801 catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct ccggactcta
     1861 gaggatcgaa cccttaaggc caccatggtg agcaagggcg aggagctgtt caccggggtg
     1921 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1981 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2041 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     2101 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     2161 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     2221 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     2281 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     2341 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     2401 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     2461 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     2521 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     2581 ggcatggacg agctgtacaa gtactcagat ctcgagctca agtagcttaa gggttcgatc
     2641 cctactggtt agtaatgagt ttaaacgggg gaggctaact gaaacacgga aggagacaat
     2701 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg
     2761 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     2821 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     2881 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag
     2941 atctgcgctg attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg
     3001 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     3061 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     3121 ctgcaggctt ggatcccaat ggcgcgccga gcttggctcg agcatggtca tagctgtttc
     3181 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3241 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     3301 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3361 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3421 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3481 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3541 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3601 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3661 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3721 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3781 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3841 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3901 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3961 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4021 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4081 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4141 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4201 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4261 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4321 ctgacagtta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
     4381 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     4441 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     4501 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     4561 tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa
     4621 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     4681 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     4741 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     4801 caggatattc ttctaatacc tggaatgctg ttttcccagg gatcgcagtg gtgagtaacc
     4861 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     4921 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     4981 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     5041 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     5101 atcgcggcct agagcaagac gtttcccgtt gaatatggct catactcttc ctttttcaat
     5161 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5221 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     5281 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5341 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     5401 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     5461 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     5521 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     5581 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     5641 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     5701 ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgacgcgc cattgggatg
     5761 ttgtaaaacg acggccagtg aa
//
