LOCUS       sequence-336847-        7047 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector carrying ADAR2-DDC-lambda-N-P2A-MCP-ADAR2-DDN with a 
            nuclear export signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7047)
  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 7047)
  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: All-in-one split-ADAR2
FEATURES             Location/Qualifiers
     source          1..7047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   96..236
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   96..225
                     /label=AAV2 ITR
     primer_bind     complement(276..292)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(276..292)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(289..311)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    300..316
                     /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(324..330,331..348,349..354))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    369..390
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(678..1266)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(758..777)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1437..2252)
                     /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(1437..2252)
                     /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     2160..2179
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2253..2357)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2425..2443
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2481..2503)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2603..2622
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2816..2838
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2831..2847
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2872..2888
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   2892..3032
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2903..3032
                     /label=AAV2 ITR
     promoter        3051..3299
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3051..3071
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3222..3241
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        3301..3375
                     /label=Cj gRNA scaffold
                     /note="guide RNA scaffold for the Campylobacter jejuni 
                     CRISPR/Cas9 system (Kim et al., 2016)"
     promoter        3460..3773
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     3692..3712
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     protein_bind    complement(3785..3809)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     enhancer        3824..4203
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4204..4407
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4358..4378
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4404..4428
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4519..4528
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5692..5718
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5725..5751
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             5761..5817
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5821..5847
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             5854..5877
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(6836..6855)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6880..6928
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgctga
       61 ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
      121 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
      181 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggcttg
      241 gatcccaatg gcgcgccgag cttggctcga gcatggtcat agctgtttcc tgtgtgaaat
      301 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
      361 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
      421 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
      481 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
      541 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
      601 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
      661 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
      721 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
      781 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
      841 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
      901 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
      961 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     1021 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     1081 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     1141 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     1201 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     1261 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     1321 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     1381 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttag
     1441 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     1501 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     1561 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     1621 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     1681 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca
     1741 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     1801 tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     1861 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     1921 tctaatacct ggaatgctgt tttcccaggg atcgcagtgg tgagtaacca tgcatcatca
     1981 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     2041 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     2101 tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta
     2161 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta
     2221 gagcaagacg tttcccgttg aatatggctc atactcttcc tttttcaata ttattgaagc
     2281 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2341 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     2401 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     2461 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     2521 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     2581 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     2641 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     2701 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     2761 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     2821 tcacgacgtt gtaaaacgac ggccagtgaa ttgacgcgcc attgggatgt tgtaaaacga
     2881 cggccagtga acctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa
     2941 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
     3001 gggagtggcc aactccatca ctaggggttc ctgcggccgc acgcgtggag gagggcctat
     3061 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattagaa
     3121 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
     3181 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
     3241 taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg
     3301 gttttagtcc ctgaaggaag ggactaaaat aaagagtttg cgggactctg cggggttaca
     3361 atcccctaaa accgctaagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
     3421 gcttttttgc tagcgtactg agtcgcccag tctcagatag atccgacgcc gccatctcta
     3481 ggcccgcgcc ggccccctcg cacagacttg tgggagaagc tcggctactc ccctgccccg
     3541 gttaatttgc atataatatt tcctagtaac tatagaggct taatgtgcga taaaagacag
     3601 ataatctgtt ctttttaata ctagctacat tttacatgat aggcttggat ttctataaga
     3661 gatacaaata ctaaattatt attttaaaaa acagcacaaa aggaaactca ccctaactgt
     3721 aaagtaattg tgtgttttga gactataaat atcccttgga gaaaagcctt gtttgaattc
     3781 ctagacccag ctttcttgta caaagttggc attaatacgc gttgacattg attattgact
     3841 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     3901 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     3961 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     4021 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     4081 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     4141 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     4201 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     4261 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     4321 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     4381 cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc
     4441 catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct ccggactcta
     4501 gaggatcgaa cccttaaggc caccatggac agacatccta ataggaaggc tagaggccaa
     4561 cttcggacga agattgaaag tggcgagggt actatcccgg tgcggtccaa cgctagtatt
     4621 caaacgtggg acggagtcct tcaaggtgaa cggctgttga caatgagctg ctcagacaaa
     4681 atcgcgcgct ggaatgtagt gggaatccaa ggcagcctct tgagcatatt cgtagaaccc
     4741 atatatttct catccattat tttgggctct ctgtatcatg gtgaccatct gtcaagggct
     4801 atgtaccaac gaatttctaa tatcgaggat cttcctccac tctatacact caataagcct
     4861 ctcttgtccg ggatatcaaa cgctgaggcc cgccagccag ggaaagctcc taacttcagt
     4921 gttaactgga ccgttggtga ttctgcgata gaggtcatca acgccacgac aggtaaggat
     4981 gagctcggta gagcctcacg cctgtgtaaa cacgcgttgt attgtagatg gatgagagta
     5041 catgggaagg tcccatctca cttgctccga agcaagatca ctaagcctaa tgtgtatcat
     5101 gagtcaaaac tcgcggctaa agaataccag gcagccaaag ctcgactttt tacagctttt
     5161 attaaggcag ggctcggggc atgggtcgag aagccgaccg agcaggacca attctctctg
     5221 acgcccgggg gcagtgggag cggtgcagga tctggtagtc cagctggggg aggagcaccg
     5281 ggtagcggtg gggggagcaa tgcgaggaca cggcgacgag aacgacgcgc tgaaaagcaa
     5341 gcgcaatgga aagctgcaaa cggaggtgga ggatctggcg gtggagggtc aggtggcggc
     5401 ggttcaaatg ctcggactcg gagacgcgag cggagagcgg agaagcaggc acaatggaaa
     5461 gccgctaatg gtggcggcgg gagtggggga ggcggatctg gaggtggcgg gtccaacgcg
     5521 agaacccgcc gacgagagcg cagagcggag aaacaagcac agtggaaggc ggccaacggg
     5581 ggtgggggat ctggcggcgg gggatccggc ggaggagggt ccaatgcccg aacccgacga
     5641 agggaaagac gggctgagaa acaggctcaa tggaaagcgg ctaatgggag ctacccatac
     5701 gatgttccag attacgctgg atccctgcca ccattggaaa gactgaccct cggatccggc
     5761 gcaacaaact tctctctgct gaaacaagcc ggagatgtcg aagagaatcc tggaccgatg
     5821 ctgcctcccc tcgaaaggct tacgctgggg agcgactaca aagacgatga cgacaagggg
     5881 agcggatccg gctctatggc gtccaatttc actcagtttg tgctggttga caacggcggg
     5941 accggggacg ttacggtagc cccctcaaac tttgccaacg gtatagcgga gtggataagc
     6001 agcaattcta ggagtcaagc atacaaagtt acatgcagcg tgcgccaatc tagcgctcag
     6061 aatcgcaagt acaccattaa agtagaggtc cccaagggag cctggagaag ctatcttaac
     6121 atggagttga ccataccaat cttcgctacc aactctgact gtgaactcat tgtgaaagcc
     6181 atgcaaggtc tgctcaagga tggtaaccca attccgtccg ctatcgctgc caactctggg
     6241 atttacgggg gcagtgggag cggtgcagga tctggtagtc cagctggggg aggagcaccg
     6301 ggtagcggtg gggggagcca gctgcacctg ccccaggttc tcgcagacgc cgtatcccgc
     6361 cttgtactgg gcaagtttgg tgatcttact gacaattttt catctcctca tgcgaggcgg
     6421 aaagtactcg caggcgtcgt catgacgacc ggaactgacg tgaaagacgc caaagtcatc
     6481 tctgtctcca cgggcacaaa gtgcataaac ggggagtaca tgagcgaccg ggggctggca
     6541 ctgaatgatt gtcacgctga aataatatct aggcgatctc tgcttagatt tctctacact
     6601 caactcgaat tgtaccttaa caacaaagat gaccagaaac gcagtatatt tcagaaatca
     6661 gaacgcggcg gatttcgact taaggaaaac gttcagttcc acttgtatat cagcacatcc
     6721 ccttgcggtg acgcccgaat cttttccccg cacgagccga tattggagga gcccgcgtga
     6781 gcttaagggt tcgatcccta ctggttagta atgagtttaa acgggggagg ctaactgaaa
     6841 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     6901 aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     6961 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     7021 ccaccccacc ccccaagttc gggtgaa
//
