LOCUS       sequence-337039-        5955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector carrying a BoxB-MS2 adRNA targeting the RAB7A transcript.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5955)
  AUTHORS   Katrekar D, Xiang Y, Palmer N, Saha A, Meluzzi D, Mali P
  TITLE     Comprehensive interrogation of the ADAR2 deaminase domain for 
            engineering enhanced RNA editing activity and specificity.
  JOURNAL   Elife. 2022 Jan 19;11. pii: 75555. doi: 10.7554/eLife.75555.
  PUBMED    35044296
REFERENCE   2  (bases 1 to 5955)
  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: "Elife."; 
            date: "2022-01-19"; volume: "11. pii: 75555. doi"; pages: " 
            10.7554/eLife.75555"
COMMENT     Sequence Label: BoxB-MS2 adRNA (RAB7A-GAC site)
FEATURES             Location/Qualifiers
     source          1..5955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   61..201
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   61..190
                     /label=AAV2 ITR
     primer_bind     complement(241..257)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(241..257)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(254..276)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    265..281
                     /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(289..295,296..313,314..319))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    334..355
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(643..1231)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(723..742)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1402..2217)
                     /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(1402..2217)
                     /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     2125..2144
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2218..2322)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2390..2408
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2446..2468)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2568..2587
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2781..2803
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2796..2812
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2837..2853
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   2857..2997
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2868..2997
                     /label=AAV2 ITR
     promoter        3016..3264
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3016..3036
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3187..3206
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        3307..3325
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        3360..3673
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     3592..3612
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     protein_bind    complement(3685..3709)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     enhancer        3724..4103
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4104..4307
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4258..4278
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4304..4328
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4419..4428
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5592..5618
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5625..5651
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(5709..5728)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5753..5801
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ccctgccata gcagatctgc gctgattttg taggtaacca cgtgcggacc gagcggccgc
       61 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
      121 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
      181 gagcgcgcag ctgcctgcag gcttggatcc caatggcgcg ccgagcttgg ctcgagcatg
      241 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
      301 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
      361 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
      421 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
      481 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
      541 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
      601 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
      661 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
      721 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
      781 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
      841 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
      901 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
      961 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1021 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1081 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1141 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     1201 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1261 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     1321 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1381 tgagtaaact tggtctgaca gttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     1441 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     1501 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     1561 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     1621 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     1681 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     1741 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     1801 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     1861 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cagggatcgc
     1921 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     1981 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     2041 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     2101 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     2161 catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcatact
     2221 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     2281 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     2341 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     2401 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     2461 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     2521 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     2581 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     2641 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     2701 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     2761 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgac
     2821 gcgccattgg gatgttgtaa aacgacggcc agtgaacctg caggcagctg cgcgctcgct
     2881 cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct
     2941 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg
     3001 gccgcacgcg tggaggaggg cctatttccc atgattcctt catatttgca tatacgatac
     3061 aaggctgtta gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa
     3121 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
     3181 aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat
     3241 atcttgtgga aaggacgaaa caccggccct gaaaaagggc caattcttgt gcctactgta
     3301 cagaaaacat gaggatcacc catgtctttt ttgcgtactg agtcgcccag tctcagatag
     3361 atccgacgcc gccatctcta ggcccgcgcc ggccccctcg cacagacttg tgggagaagc
     3421 tcggctactc ccctgccccg gttaatttgc atataatatt tcctagtaac tatagaggct
     3481 taatgtgcga taaaagacag ataatctgtt ctttttaata ctagctacat tttacatgat
     3541 aggcttggat ttctataaga gatacaaata ctaaattatt attttaaaaa acagcacaaa
     3601 aggaaactca ccctaactgt aaagtaattg tgtgttttga gactataaat atcccttgga
     3661 gaaaagcctt gtttgaattc ctagacccag ctttcttgta caaagttggc attaatacgc
     3721 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     3781 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     3841 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     3901 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     3961 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     4021 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     4081 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     4141 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     4201 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     4261 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc
     4321 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc
     4381 gatccagcct ccggactcta gaggatcgaa cccttaaggc caccatggac agacatccta
     4441 ataggaaggc tagaggccaa cttcggacga agattgaaag tggccagggt actatcccgg
     4501 tgcggtccaa cgctagtatt caaacgtggg acggagtcct tcaaggtgaa cggctgttga
     4561 caatgagctg ctcagacaaa atcgcgcgct ggaatgtagt gggaatccaa ggcagcctct
     4621 tgagcatatt cgtagaaccc atatatttct catccattat tttgggctct ctgtatcatg
     4681 gtgaccatct gtcaagggct atgtaccaac gaatttctaa tatcgaggat cttcctccac
     4741 tctatacact caataagcct ctcttgtccg ggatatcaaa cgctgaggcc cgccagccag
     4801 ggaaagctcc taacttcagt gttaactgga ccgttggtga ttctgcgata gaggtcatca
     4861 acgccacgac aggtaaggat gagctcggta gagcctcacg cctgtgtaaa cacgcgttgt
     4921 attgtagatg gatgagagta catgggaagg tcccatctca cttgctccga agcaagatca
     4981 ctaagcctaa tgtgtatcat gagtcaaaac tcgcggctaa agaataccag gcagccaaag
     5041 ctcgactttt tacagctttt attaaggcag ggctcggggc atgggtcgag aagccgaccg
     5101 agcaggacca attctctctg acgcccgggg gcagtgggag cggtgcagga tctggtagtc
     5161 cagctggggg aggagcaccg ggtagcggtg gggggagcaa tgcgaggaca cggcgacgag
     5221 aacgacgcgc tgaaaagcaa gcgcaatgga aagctgcaaa cggaggtgga ggatctggcg
     5281 gtggagggtc aggtggcggc ggttcaaatg ctcggactcg gagacgcgag cggagagcgg
     5341 agaagcaggc acaatggaaa gccgctaatg gtggcggcgg gagtggggga ggcggatctg
     5401 gaggtggcgg gtccaacgcg agaacccgcc gacgagagcg cagagcggag aaacaagcac
     5461 agtggaaggc ggccaacggg ggtgggggat ctggcggcgg gggatccggc ggaggagggt
     5521 ccaatgcccg aacccgacga agggaaagac gggctgagaa acaggctcaa tggaaagcgg
     5581 ctaatgggag ctacccatac gatgttccag attacgctgg atccctgcca ccattggaaa
     5641 ggcttacgct gtgacataag ggttcgatcc ctactggtta gtaatgagtt taaacggggg
     5701 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     5761 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     5821 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     5881 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     5941 cgtcggggcg gcagg
//
