LOCUS       sequence-422994-        5675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses acyclovir-controllable IRE variant #A8 as circular RNA in 
            mammalian cells using the Tornado expression system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5675)
  AUTHORS   Hagen T, Litke JL, Nasir N, Hou Q, Jaffrey SR
  TITLE     Engineering acyclovir-induced RNA nanodevices for reversible and 
            tunable control of aptamer function.
  JOURNAL   Cell Chem Biol. 2024 Aug 23:S2451-9456(24)00319-2. doi: 
            10.1016/j.chembiol.2024.07.017.
  PUBMED    39191249
REFERENCE   2  (bases 1 to 5675)
  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: "Cell Chem 
            Biol. 2024 Aug 23:S2451-9456(24)00319-2. doi: 
            10.1016/j.chembiol.2024.07.017."
COMMENT     Sequence Label: pAV-puro-U6+27-F30-Tornado-Acyclovir-IRE#A8
FEATURES             Location/Qualifiers
     source          1..5675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..273
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     25..45
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     196..215
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    660..781
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(878..1333)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1015..1036)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1227..1246
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1360..1464
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1465..1533,1534..2325)
                     /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(1683..1702)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2496..3084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2985..3004
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3527..3661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4086..4106)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4236..4301
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4729..5328)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4812..4832)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5309..5328
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(5338..5667)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5352..5487
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5652..5672
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 gccggatcca aggtcgggca ggaagagggc ctatttccca tgattccttc atatttgcat
       61 atacgataca aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata
      121 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
      181 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
      241 gctttatata tcttgtggaa aggacgaaac accgtgctcg cttcggcagc acatatacta
      301 gtcgacgggc cgcactcgcc ggtcccaagc ccggataaaa tgggaggggg cgggaaaccg
      361 cctaaccatg ccgagtgcgg ccgcttgcca tgtgctgctt caacagtgct tggacgctct
      421 gatgaataac cgaagtagtt tatgggctac cgaaatcatt catggcaagt ggccgcggtc
      481 ggcgtggact gtagaacact gccaatgccg gtcccaagcc cggataaaag tggagggtac
      541 agtccacgct ctagagcgca cttcggtgcg ctttttacta ggacctgcag gcatgcaagc
      601 ttgacgtcgg ttaccgatat ccatatggcg accgcatcga tctcgagccg aggactagta
      661 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
      721 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
      781 atcatgtctt acgtagataa gtagcatggc gggttatagc gaagaggccc gcaccgatcg
      841 cccttcccaa cagttgcgca gcctgaatgg cgaatggaaa ttgtaagcgt taatattttg
      901 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
      961 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     1021 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     1081 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     1141 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     1201 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     1261 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     1321 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     1381 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     1441 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     1501 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     1561 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     1621 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     1681 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     1741 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     1801 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     1861 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     1921 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     1981 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     2041 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     2101 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2161 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     2221 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2281 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     2341 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     2401 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     2461 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2521 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2581 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2641 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2701 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2761 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2821 ggttcgtgca acacagccag cttggagcga acgacctaca ccgaactgag atacctacag
     2881 cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2941 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3001 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3061 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3121 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     3181 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     3241 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     3301 tggccgattc attaatgcag agatctttgg ccactccctc tctgcgcgct cgctcgctca
     3361 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3421 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc tggaggggtg
     3481 gagtcgtgac gtgaattacg tcatagggtt agggaggtcc tggatcgatc cagacatgat
     3541 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     3601 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3661 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     3721 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatctct agtcaaggca
     3781 ctatacatca aatattcctt attaacccct ttacaaatta aaaagctaaa ggtacacaat
     3841 ttttgagcat agttattaat agcagacact ctatgcctgt gtggagtaag aaaaaacagt
     3901 atgttatgat tataactgtt atgcctactt ataaaggtta cagaatattt ttccataatt
     3961 ttcttgtata gcagtgcagc tttttccttt gtggtgtaaa tagcaaagca agcaagagtt
     4021 ctattactaa acacagcatg actcaaaaaa cttagcaatt ctgaaggaaa gtccttgggg
     4081 tcttctacct ttctcttctt ttttggagga gtagaatgtt gagagtcagc agtagcctca
     4141 tcatcactag atggcatttc ttctgagcaa aacaggtttt cctcattaaa ggcattccac
     4201 cactgctccc attcatcagt tccataggtt ggaatctaaa atacacaaac aattagaatc
     4261 agtagtttaa cacattatac acttaaaaat tttatattta ccttagagct ttaaatctct
     4321 gtaggtagtt tgtccaatta tgtcacacca cagaagtaag gttccttcac aaagatccgg
     4381 gaccaaagcg gccatcgtgc ctccccactc ctgcagttcg ggggcatgga tgcgcggata
     4441 gccgctgctg gtttcctgga tgccgacgga tttgcactgc cggtagaact ccgcgaggtc
     4501 gtccagcctc aggcagcagc tgaaccaact cgcgagggga tcgagcccgg ggtgggcgaa
     4561 gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat
     4621 tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt
     4681 gggcgtcgct tggtcggtca tttcgaactg cgggtcgtgg ggcgggcgtc aggcaccggg
     4741 cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt
     4801 gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg
     4861 caccccggcg cgctcggccg cctccactcc ggggagcacg acggcgctgc ccagaccctt
     4921 gccctggtgg tcgggcgaga cgccgacggt ggccaggaac cacgcgggct ccttgggccg
     4981 gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc agccgggaac cgctcaactc
     5041 ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt
     5101 ccagaccgcc acggcagcac cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac
     5161 gcgcgtgagg aagagttctt gcagctcggt gacccgctcg atgtggcgat caggatcgac
     5221 ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggccctggg
     5281 gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg taagcttttt
     5341 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     5401 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     5461 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     5521 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     5581 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     5641 ggactttcca caccctaact gacacacatt ccaca
//
