LOCUS       sequence-336846-        5681 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector carrying a guide RNA targeting the human RAB7A mRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5681)
  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 5681)
  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 100,50 (RAB7A)
FEATURES             Location/Qualifiers
     source          1..5681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3..25)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14..30
                     /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(38..44,45..62,63..68))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    83..104
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(392..980)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(472..491)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1151..1966)
                     /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(1151..1966)
                     /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     1874..1893
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1967..2071)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2139..2157
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2195..2217)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2317..2336
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2530..2552
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2545..2561
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2586..2602
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   2606..2746
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2617..2746
                     /label=AAV2 ITR
     promoter        2765..3013
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2765..2785
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2936..2955
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        3324..3637
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     3556..3576
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     protein_bind    complement(3649..3673)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     enhancer        3688..4067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4068..4271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4222..4242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4268..4292
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4383..4392
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4389..4391,4392..4394,4395..5105)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4389..4391,4392..4394,4395..5105)
                     /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(4434..4455)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4695..4714)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5042..5063
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5184..5203)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5228..5276
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     repeat_region   5491..5631
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5491..5620
                     /label=AAV2 ITR
     primer_bind     complement(join(5671..5681,1..6))
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(join(5671..5681,1..6))
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
       61 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
      121 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
      181 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
      241 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
      301 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
      361 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
      421 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
      481 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
      541 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
      601 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
      661 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
      721 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
      781 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
      841 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
      901 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
      961 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     1021 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     1081 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     1141 ggtctgacag ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     1201 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     1261 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     1321 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     1381 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
     1441 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     1501 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
     1561 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     1621 aatcaggata ttcttctaat acctggaatg ctgttttccc agggatcgca gtggtgagta
     1681 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     1741 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     1801 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     1861 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     1921 ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcatactc ttcctttttc
     1981 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     2041 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     2101 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     2161 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     2221 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     2281 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     2341 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     2401 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     2461 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     2521 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgacg cgccattggg
     2581 atgttgtaaa acgacggcca gtgaacctgc aggcagctgc gcgctcgctc gctcactgag
     2641 gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
     2701 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgcacgcgt
     2761 ggaggagggc ctatttccca tgattccttc atatttgcat atacgataca aggctgttag
     2821 agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg
     2881 tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta
     2941 tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa
     3001 aggacgaaac accgccatca gtcgccggtc ccaagcccgg ataaaatggg agggggcggg
     3061 aaaccgccta accatgccga ctgatggcag aaaaaaaaaa tgataaaagg cgtacataat
     3121 tcttgtgtct actgtacaga atactgccgc cagctggatt tcccaattct gagtaacact
     3181 ctgcaatcca aacagggttc aaaaaaaaaa ctgccatcag tcggcgtgga ctgtagaaca
     3241 ctgccaatgc cggtcccaag cccggataaa agtggagggt acagtccacg cttttttcgt
     3301 actgagtcgc ccagtctcag atagatccga cgccgccatc tctaggcccg cgccggcccc
     3361 ctcgcacaga cttgtgggag aagctcggct actcccctgc cccggttaat ttgcatataa
     3421 tatttcctag taactataga ggcttaatgt gcgataaaag acagataatc tgttcttttt
     3481 aatactagct acattttaca tgataggctt ggatttctat aagagataca aatactaaat
     3541 tattatttta aaaaacagca caaaaggaaa ctcaccctaa ctgtaaagta attgtgtgtt
     3601 ttgagactat aaatatccct tggagaaaag ccttgtttga attcctagac ccagctttct
     3661 tgtacaaagt tggcattaat acgcgttgac attgattatt gactagttat taatagtaat
     3721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     3781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     3841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     3901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     3961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     4021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     4081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     4141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     4201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     4261 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
     4321 acctccatag aagacaccgg gaccgatcca gcctccggac tctagaggat cgaaccctta
     4381 aggccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     4441 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     4501 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     4561 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     4621 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     4681 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     4741 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     4801 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     4861 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     4921 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     4981 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     5041 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     5101 acaagtactc agatctcgag ctcaagtagc ttaagggttc gatccctact ggttagtaat
     5161 gagtttaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     5221 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     5281 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5341 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5401 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gctgattttg
     5461 taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     5521 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     5581 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gcttggatcc
     5641 caatggcgcg ccgagcttgg ctcgagcatg gtcatagctg t
//
