LOCUS       sequence-382107-        7529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ADAR2, sesRNA in mammalian cells, with tTA2 as efRNA 
            and ADAR2 overexpression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7529)
  AUTHORS   Qian Y, Li J, Zhao S, Matthews EA, Adoff M, Zhong W, An X, Yeo M, 
            Park C, Yang X, Wang BS, Southwell DG, Huang ZJ
  TITLE     Programmable RNA sensing for cell monitoring and manipulation.
  JOURNAL   Nature. 2022 Oct;610(7933):713-721. doi: 10.1038/s41586-022-05280-1.
            Epub 2022 Oct 5.
  PUBMED    36198803
REFERENCE   2  (bases 1 to 7529)
  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"; doi: 
            "10.1038/s41586-022-05280-1"; journalName: "Nature"; date: 
            "2022-10"; volume: "610"; issue: "7933"; pages: "713-721"
COMMENT     Sequence Label: CAG-ADAR2-sesRNA-GFP-W3SL
FEATURES             Location/Qualifiers
     source          1..7529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      97..552
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(184..203)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     394..415
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(569..588)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     688..710
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(748..766)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        834..938
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(939..1007,1008..1799)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1157..1176)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1970..2558
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2459..2478
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2631..2760
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        2775..3060
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          3285..3512
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             3533..3559
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5666..5722
                     /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             6149..6202
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(6212..6214,6215..6928)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(6212..6214,6215..6925)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6254..6275)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6515..6534)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6862..6883
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7003..7023)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   join(7411..7529,1..22)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   join(7411..7529,1..11)
                     /label=AAV2 ITR
ORIGIN
        1 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
       61 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
      121 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
      181 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
      241 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
      301 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
      361 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
      421 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
      481 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
      541 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
      601 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
      661 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
      721 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
      781 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
      841 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
      901 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
      961 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     1021 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     1081 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     1141 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     1201 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     1261 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     1321 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     1381 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     1441 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     1501 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     1561 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     1621 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     1681 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     1741 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     1801 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     1861 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     1921 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1981 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2041 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2101 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2161 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2221 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2281 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2341 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2401 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2461 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2521 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2581 tacggttcct ggccttttgc tggccttttg ctggcctttt gctcacatgt cctgcaggca
     2641 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
     2701 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
     2761 gcggccgcac gcgtcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     2821 gacccccgcc cattgacgtc aatagtaacg ccaataggga ctttccattg acgtcaatgg
     2881 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     2941 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattgtgc ccagtacatg
     3001 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     3061 gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca
     3121 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg gcgaggcgga
     3181 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
     3241 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct
     3301 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
     3361 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
     3421 tgagcaagag gtaagggttt aagggatggt tggttggtgg ggtattaatg tttaattacc
     3481 tggagcacct gcctgaaatc actttttttc aggttgggga tccgccacca tgtacccata
     3541 cgatgttcca gattacgctg gaggcgatat agaagatgaa gaaaacatga gttccagcag
     3601 cactgatgtg aaggaaaacc gcaatctgga caacgtgtcc cccaaggatg gcagcacacc
     3661 tgggcctggc gagggctctc agctctccaa tgggggtggt ggtggccccg gcagaaagcg
     3721 gcccctggag gagggcagca atggccactc caagtaccgc ctgaagaaaa ggaggaaaac
     3781 accagggccc gtcctcccca agaacgccct gatgcagctg aatgagatca agcctggttt
     3841 gcagtacaca ctcctgtccc agactgggcc cgtgcacgcg cctttgtttg tcatgtctgt
     3901 ggaggtgaat ggccaggttt ttgagggctc tggtcccaca aagaaaaagg caaaactcca
     3961 tgctgctgag aaggccttga ggtctttcgt tcagtttact aatgcctctg aggcccacct
     4021 ggccatgggg aggaccctgt ctgtcaacac ggacttcaca tctgaccagg ccgacttccc
     4081 tgacacgctc ttcaatggtt ttgaaactcc tgacaaggcg gagcctccct tttacgtggg
     4141 ctccaatggg gatgactcct tcagttccag cggggacctc agcttgtctg cttccccggt
     4201 gcctgccagc ctagcccagc ctcctctccc tgtcttacca ccattcccac ccccgagtgg
     4261 gaagaatccc gtgatgatct tgaacgaact gcgcccagga ctcaagtatg acttcctctc
     4321 cgagagcggg gagagccatg ccaagagctt cgtcatgtct gtggtcgtgg atggtcagtt
     4381 ctttgaaggc tcggggagaa acaagaagct tgccaaggcc cgggctgcgc agtctgccct
     4441 ggccgccatt tttaacttgc acttggatca gacgccatct cgccagccta ttcccagtga
     4501 gggtcttcag ctgcatttac cgcaggtttt agctgacgct gtctcacgcc tggtcctggg
     4561 taagtttggt gacctgaccg acaacttctc ctcccctcac gctcgcagaa aagtgctggc
     4621 tggagtcgtc atgacaacag gcacagatgt taaagatgcc aaggtgataa gtgtttctac
     4681 aggaacaaaa tgtattaatg gtgaatacat gagtgatcgt ggccttgcat taaatgactg
     4741 ccatgcagaa ataatatctc ggagatcctt gctcagattt ctttatacac aacttgagct
     4801 ttacttaaat aacaaagatg atcaaaaaag atccatcttt cagaaatcag agcgaggggg
     4861 gtttaggctg aaggagaatg tccagtttca tctgtacatc agcacctctc cctgtggaga
     4921 tgccagaatc ttctcaccac atgagccaat cctggaagaa ccagcagata gacacccaaa
     4981 tcgtaaagca agaggacagc tacggaccaa aatagagtct ggtgagggga cgattccagt
     5041 gcgctccaat gcgagcatcc aaacgtggga cggggtgctg caaggggagc ggctgctcac
     5101 catgtcctgc agtgacaaga ttgcacgctg gaacgtggtg ggcatccagg gatccctgct
     5161 cagcattttc gtggagccca tttacttctc gagcatcatc ctgggcagcc tttaccacgg
     5221 ggaccacctt tccagggcca tgtaccagcg gatctccaac atagaggacc tgccacctct
     5281 ctacaccctc aacaagcctt tgctcagtgg catcagcaat gcagaagcac ggcagccagg
     5341 gaaggccccc aacttcagtg tcaactggac ggtaggcgac tccgctattg aggtcatcaa
     5401 cgccacgact gggaaggatg agctgggccg cgcgtcccgc ctgtgtaagc acgcgttgta
     5461 ctgtcgctgg atgcgtgtgc acggcaaggt tccctcccac ttactacgct ccaagattac
     5521 caaacccaac gtgtaccatg agtccaagct ggcggcaaag gagtaccagg ccgccaaggc
     5581 gcgtctgttc acagccttca tcaaggcggg gctgggggcc tgggtggaga agcccaccga
     5641 gcaggaccag ttctcactca cgcccgctac taacttcagc ctgctgaagc aggctggaga
     5701 cgtggaggag aaccctggac ctgaattctt ccgacgatgt ggcgaaaggc gactggcgcg
     5761 catccgtgaa gcccaggaat gggtccttca tgaagtggcg cgatgctgcg tagcccacga
     5821 gccactgcga gtacacgttc tcagatagga tgagggcggc aggtagcagc tccaggtctt
     5881 tctccacctt gatgcgcttg gccgcgtgca gcgcgggacc accgagccca gggcttttgt
     5941 ggtcatcttc atcaggggtg acctggatcc cgatctccac tggctcagat actctcctga
     6001 gctcagagcg agaggaggga ggtagactgc tcttgtccag gaccttgtcg tagcaggggc
     6061 ccaggcctcc acactgtttc ttcttgtgct ctataaaaac caggatgtcc cccagcggga
     6121 agttcatctg acactggccg gcgcgccaga gggcagagga agtcttctaa catgcggtga
     6181 cgtggaggag aatcccggcc ctttaattaa cgtgagcaag ggcgaggagc tgttcaccgg
     6241 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     6301 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     6361 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     6421 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     6481 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     6541 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     6601 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     6661 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     6721 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     6781 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     6841 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     6901 tctcggcatg gacgagctgt acaagtaagc tagcaattcc gctcgagata atcaacctct
     6961 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     7021 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     7081 tttctcctcc ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg
     7141 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgtttat
     7201 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatc tagctttatt tgtgaaattt
     7261 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     7321 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcggcc
     7381 gcggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     7441 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     7501 gctttgcccg ggcggcctca gtgagcgag
//
