LOCUS       sequence-381702-        7205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ADAR2, sesRNA in mammalian cells with hSyn promoter, 
            with tTA2 as efRNA and ADAR2 overexpression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7205)
  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 7205)
  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: pAAV-hSyn-ADAR2-sesRNA-tTA2-W3SL
FEATURES             Location/Qualifiers
     source          1..7205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..448
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      468..477
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2577..2633
                     /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             3033..3086
                     /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(3096..3710,3711..3716,3717..3752,3753..3755,3756..3791,
                     3792..3794,3795..3830,3831..3839)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=tTA-Advanced
                     /note="In the Tet-Off(R) system, tTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="SRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVK
                     NKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRP
                     TEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTD
                     SMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPAD
                     ALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(3914..3934)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4294..4434
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4294..4423
                     /label=AAV2 ITR
     rep_origin      4509..4964
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4596..4615)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4806..4827
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4981..5000)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5100..5122
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5160..5178)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5246..5350
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5351..5419,5420..6211)
                     /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(5569..5588)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6382..6970
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6871..6890
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   7032..7161
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
       61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      121 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      421 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggtac cggatccgcc accatggata
      481 tagaagatga agaaaacatg agttccagca gcactgatgt gaaggaaaac cgcaatctgg
      541 acaacgtgtc ccccaaggat ggcagcacac ctgggcctgg cgagggctct cagctctcca
      601 atgggggtgg tggtggcccc ggcagaaagc ggcccctgga ggagggcagc aatggccact
      661 ccaagtaccg cctgaagaaa aggaggaaaa caccagggcc cgtcctcccc aagaacgccc
      721 tgatgcagct gaatgagatc aagcctggtt tgcagtacac actcctgtcc cagactgggc
      781 ccgtgcacgc gcctttgttt gtcatgtctg tggaggtgaa tggccaggtt tttgagggct
      841 ctggtcccac aaagaaaaag gcaaaactcc atgctgctga gaaggccttg aggtctttcg
      901 ttcagtttcc taatgcctct gaggcccacc tggccatggg gaggaccctg tctgtcaaca
      961 cggacttcac atctgaccag gccgacttcc ctgacacgct cttcaatggt tttgaaactc
     1021 ctgacaaggc ggagcctccc ttttacgtgg gctccaatgg ggatgactcc ttcagttcca
     1081 gcggggacct cagcttgtct gcttccccgg tgcctgccag cctagcccag cctcctctcc
     1141 ctgtcttacc accattccca cccccgagtg ggaagaatcc cgtgatgatc ttgaacgaac
     1201 tgcgcccagg actcaagtat gacttcctct ccgagagcgg ggagagccat gccaagagct
     1261 tcgtcatgtc tgtggtcgtg gatggtcagt tctttgaagg ctcggggaga aacaagaagc
     1321 ttgccaaggc ccgggctgcg cagtctgccc tggccgccat ttttaacttg cacttggatc
     1381 agacgccatc tcgccagcct attcccagtg agggtcttca gctgcattta ccgcaggttt
     1441 tagctgacgc tgtctcacgc ctggtcctgg gtaagtttgg tgacctgacc gacaacttct
     1501 cctcccctca cgctcgcaga aaagtgctgg ctggagtcgt catgacaaca ggcacagatg
     1561 ttaaagatgc caaggtgata agtgtttcta caggaacaaa atgtattaat ggtgaataca
     1621 tgagtgatcg tggccttgca ttaaatgact gccatgcaga aataatatct cggagatcct
     1681 tgctcagatt tctttataca caacttgagc tttacttaaa taacaaagat gatcaaaaaa
     1741 gatccatctt tcagaaatca gagcgagggg ggtttaggct gaaggagaat gtccagtttc
     1801 atctgtacat cagcacctct ccctgtggag atgccagaat cttctcacca catgagccaa
     1861 tcctggaaga accagcagat agacacccaa atcgtaaagc aagaggacag ctacggacca
     1921 aaatagagtc tggtgagggg acgattccag tgcgctccaa tgcgagcatc caaacgtggg
     1981 acggggtgct gcaaggggag cggctgctca ccatgtcctg cagtgacaag attgcacgct
     2041 ggaacgtggt gggcatccag ggatccctgc tcagcatttt cgtggagccc atttacttct
     2101 cgagcatcat cctgggcagc ctttaccacg gggaccacct ttccagggcc atgtaccagc
     2161 ggatctccaa catagaggac ctgccacctc tctacaccct caacaagcct ttgctcagtg
     2221 gcatcagcaa tgcagaagca cggcagccag ggaaggcccc caacttcagt gtcaactgga
     2281 cggtaggcga ctccgctatt gaggtcatca acgccacgac tgggaaggat gagctgggcc
     2341 gcgcgtcccg cctgtgtaag cacgcgttgt actgtcgctg gatgcgtgtg cacggcaagg
     2401 ttccctccca cttactacgc tccaagatta ccaagcccaa cgtgtaccat gagtccaagc
     2461 tggcggcaaa ggagtaccag gccgccaagg cgcgtctgtt cacagccttc atcaaggcgg
     2521 ggctgggggc ctgggtggag aagcccaccg agcaggacca gttctcactc acgcccgcta
     2581 ctaacttcag cctgctgaag caggctggag acgtggagga gaaccctgga cctgaattcg
     2641 aagcgcaggc tgccgttctc agaggaatgt tccgacgatg tggcgaaagg cgactggcgc
     2701 gcatccgtga agcccaggaa tgggtccttc atgaagtggc gcgatgctgc gtagcccacg
     2761 agccactgcg agtacacgtt ctcagatggg atgagggcgg caggtagcag ctccaggtct
     2821 ttctccacct tgatgcgctt ggccgcgtgc agcgcgggac caccgagccc agggctgggc
     2881 agcggtgctg gcttgcgtgg gaagagagct ggaaagggct ctgcgcctgg cgcgaaggcc
     2941 ccgccgcgcc cgttcactgc acccggtgca cccgcgtccc cacagccaac agctccggca
     3001 tcacccgtgt cgcctgcacg cttggcgcgc cagagggcag aggaagtctt ctaacatgcg
     3061 gtgacgtgga ggagaatccc ggccctttaa ttaactctag actggacaag agcaaagtca
     3121 taaactctgc tctggaatta ctcaatgaag tcggtatcga aggcctgacg acaaggaaac
     3181 tcgctcaaaa gctgggagtt gagcagccta ccctgtactg gcacgtgaag aacaagcggg
     3241 ccctgctcga tgccctggca atcgagatgc tggacaggca tcatacccac ttctgccccc
     3301 tggaaggcga gtcatggcaa gactttctgc ggaacaacgc caagtcattc cgctgtgctc
     3361 tcctctcaca tcgcgacggg gctaaagtgc atctcggcac ccgcccaaca gagaaacagt
     3421 acgaaaccct ggaaaatcag ctcgcgttcc tgtgtcagca aggcttctcc ctggagaacg
     3481 cactgtacgc tctgtccgcc gtgggccact ttacactggg ctgcgtattg gaggatcagg
     3541 agcatcaagt agcaaaagag gaaagagaga cacctaccac cgattctatg cccccacttc
     3601 tgagacaagc aattgagctg ttcgaccatc agggagccga acctgccttc cttttcggcc
     3661 tggaactaat catatgtggc ctggaaaaac agctaaagtg cgaaagcggc ggaccggccg
     3721 acgcccttga cgattttgac ttagacatgc tcccagccga tgcccttgac gactttgacc
     3781 ttgatatgct gcctgctgac gctcttgacg attttgacct tgacatgctc cccgggtaag
     3841 ctagcaattc cgctcgagat aatcaacctc tggattacaa aatttgtgaa agattgactg
     3901 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     3961 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag
     4021 ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     4081 tgttgggcac tgacaattcc gtggtgttta tttgtgaaat ttgtgatgct attgctttat
     4141 ttgtaaccat ctagctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     4201 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     4261 ggggagatgt gggaggtttt ttaaagcggc cgcaggaacc cctagtgatg gagttggcca
     4321 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     4381 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     4441 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     4501 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4561 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     4621 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4681 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     4741 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4801 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     4861 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     4921 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     4981 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     5041 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5101 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     5161 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     5221 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5281 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5341 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5401 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5461 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5521 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5581 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5641 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5701 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5761 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5821 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5881 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5941 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6001 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6061 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6121 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6181 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6241 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6301 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6361 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6421 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6481 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6541 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6601 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6661 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6721 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6781 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6841 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6901 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6961 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7021 tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc
     7081 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
     7141 actccatcac taggggttcc tgcggccgca cgcgtgtgtc tagactgcag agggccctgc
     7201 gtatg
//
