LOCUS       sequence-356528-        6452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector carrying the MCP-ADAR2-DD(E488Q, N496F) with a nuclear 
            export signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6452)
  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 6452)
  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: MCP-ADAR2-DD(E488Q, N496F)
FEATURES             Location/Qualifiers
     source          1..6452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    520..541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(556..561,562..579,580..586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    594..610
                     /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)."
     primer_bind     599..621
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     618..634
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     618..634
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   674..814
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   685..814
                     /label=AAV2 ITR
     polyA_signal    1029..1077
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1102..1121
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2778..2801)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(2808..2834)
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(2934..2958)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2955..3158)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2984..3004)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3159..3538
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3553..3577
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(3589..3902)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     complement(3650..3670)
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     misc_RNA        complement(3987..4061)
                     /label=Cj gRNA scaffold
                     /note="guide RNA scaffold for the Campylobacter jejuni 
                     CRISPR/Cas9 system (Kim et al., 2016)"
     promoter        complement(4063..4311)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(4121..4140)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(4291..4311)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     repeat_region   4330..4470
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4330..4459
                     /label=AAV2 ITR
     primer_bind     complement(4474..4490)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4515..4531)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4524..4546)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4740..4759)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4859..4881
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4919..4937)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5005..5109
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5110..5925
                     /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             5110..5925
                     /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     complement(5183..5202)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(6096..6452,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      541 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      601 cggataacaa tttcacacag gaaacagcta tgaccatgct cgagccaagc tcggcgcgcc
      661 attgggatcc aagcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca
      721 aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
      781 gagggagtgg ccaactccat cactaggggt tcctgcggcc gctcggtccg cacgtggtta
      841 cctacaaaat cagcgcagat ctgctatggc agggcctgcc gccccgacgt tggctgcgag
      901 ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta
      961 ttggccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc
     1021 gcgtttatga acaaacgacc caacacccgt gcgttttatt ctgtcttttt attgccgtca
     1081 tagcgcgggt tccttccggt attgtctcct tccgtgtttc agttagcctc ccccgtttaa
     1141 actcattact aaccagtagg gatcgaaccc ttaagtcagg gcgtcagaga gaattggtcc
     1201 tgctcggtcg gcttctcgac ccatgccccg agccctgcct taataaaagc tgtaaaaagt
     1261 cgagctttgg ctgcctggta ttctttagcc gcgagttttg actcatgata cacattaggc
     1321 ttagtgatct tgcttcggag caagtgagat gggaccttcc catgtactct catccatcta
     1381 caatacaacg cgtgtttaca caggcgtgag gctctaccga gctcatcctt acctgtcgtg
     1441 gcgttgatga cctctatcgc agaatcacca acggtccagt taacactgaa gttaggagct
     1501 ttccctggct ggcgggcctc agcgtttgat atcccggaca agagaggctt attgagtgta
     1561 tagagtggag gaagatcctc gatattagaa attcgttggt acatagccct tgacagatgg
     1621 tcaccatgat acagagagcc caaaataatg gatgagaaat atatgggttc tacgaatatg
     1681 ctcaagaggc tgccttggat tcccactaca ttccagcgcg cgattttgtc tgagcagctc
     1741 attgtcaaca gccgttcacc ttgaaggact ccgtcccacg tttgaatact agcgaaggac
     1801 cgcaccggga tagtaccctg gccactttca atcttcgtcc gaagttggcc tctagccttc
     1861 ctattaggat gtctgtccgc gggctcctcc aatatcggct cgtgcgggga aaagattcgg
     1921 gcgtcaccgc aaggggatgt gctgatatac aagtggaact gaacgttttc cttaagtcga
     1981 aatccgccgc gttctgattt ctgaaatata ctgcgtttct ggtcatcttt gttgttaagg
     2041 tacaattcga gttgagtgta gagaaatcta agcagagatc gcctagatat tatttcagcg
     2101 tgacaatcat tcagtgccag cccccggtcg ctcatgtact ccccgtttat gcactttgtg
     2161 cccgtggaga cagagatgac tttggcgtct ttcacgtcag ttccggtcgt catgacgacg
     2221 cctgcgagta ctttccgcct cgcatgagga gatgaaaaat tgtcagtaag atcaccaaac
     2281 ttgcccagta caaggcggga tacggcgtct gcgagaacct ggggcaggtg cagctgagac
     2341 cccccaccgc tacccggtgc tcctccccca gctggactac cagatcctgc accgctccca
     2401 ctgcccccgt aaatcccaga gttggcagcg atagcggacg gaattgggtt accatccttg
     2461 agcagacctt gcatggcttt cacaatgagt tcacagtcag agttggtagc gaagattggt
     2521 atggtcaact ccatgttaag atagcttctc caggctccct tggggacctc tactttaatg
     2581 gtgtacttgc gattctgagc gctagattgg cgcacgctgc atgtaacttt gtatgcttga
     2641 ctcctagaat tgctgcttat ccactccgct ataccgttgg caaagtttga gggggctacc
     2701 gtaacgtccc cggtcccgcc gttgtcaacc agcacaaact gagtgaaatt ggacgccata
     2761 gagccggatc cgctcccctt gtcgtcatcg tctttgtagt cgctccccag cgtaagcctt
     2821 tccaatggtg gcagcatggt ggccttaagg gttcgatcct ctagagtccg gaggctggat
     2881 cggtcccggt gtcttctatg gaggtcaaaa cagcgtggat ggcgtctcca ggcgatctga
     2941 cggttcacta aacgagctct gcttatatag acctcccacc gtacacgcct accgcccatt
     3001 tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa
     3061 aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc
     3121 tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat
     3181 acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag
     3241 gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga
     3301 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
     3361 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
     3421 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
     3481 ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcaa
     3541 cgcgtattaa tgccaacttt gtacaagaaa gctgggtcta ggaattcaaa caaggctttt
     3601 ctccaaggga tatttatagt ctcaaaacac acaattactt tacagttagg gtgagtttcc
     3661 ttttgtgctg ttttttaaaa taataattta gtatttgtat ctcttataga aatccaagcc
     3721 tatcatgtaa aatgtagcta gtattaaaaa gaacagatta tctgtctttt atcgcacatt
     3781 aagcctctat agttactagg aaatattata tgcaaattaa ccggggcagg ggagtagccg
     3841 agcttctccc acaagtctgt gcgagggggc cggcgcgggc ctagagatgg cggcgtcgga
     3901 tctatctgag actgggcgac tcagtacgct agcaaaaaag caccgactcg gtgccacttt
     3961 ttcaagttga taacggacta gccttagcgg ttttagggga ttgtaacccc gcagagtccc
     4021 gcaaactctt tattttagtc ccttccttca gggactaaaa ccggtgtttc gtcctttcca
     4081 caagatatat aaagccaaga aatcgaaata ctttcaagtt acggtaagca tatgatagtc
     4141 cattttaaaa cataatttta aaactgcaaa ctacccaaga aattattact ttctacgtca
     4201 cgtattttgt actaatatct ttgtgtttac agtcaaatta attctaatta tctctctaac
     4261 agccttgtat cgtatatgca aatatgaagg aatcatggga aataggccct cctccacgcg
     4321 tgcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     4381 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     4441 tgagcgagcg agcgcgcagc tgcctgcagg ttcactggcc gtcgttttac aacatcccaa
     4501 tggcgcgtca attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     4561 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     4621 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     4681 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     4741 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     4801 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4861 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     4921 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4981 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5041 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5101 aggaagagta tgagccatat tcaacgggaa acgtcttgct ctaggccgcg attaaattcc
     5161 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     5221 gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     5281 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     5341 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     5401 accactgcga tccctgggaa aacagcattc caggtattag aagaatatcc tgattcaggt
     5461 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     5521 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     5581 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     5641 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     5701 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     5761 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     5821 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     5881 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaactgtc agaccaagtt
     5941 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6001 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6061 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6121 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6181 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6241 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6301 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6361 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6421 ggttcgtgca cacagcccag cttggagcga ac
//
