LOCUS       sequence-430576-        5500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses acyclovir-controllable IRE variant #A8-4 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 5500)
  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 5500)
  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#A8-4
FEATURES             Location/Qualifiers
     source          1..5500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     285..305
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        333..581
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     333..353
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     504..523
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    968..1089
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1186..1641)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1323..1344)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1535..1554
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1668..1772
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1773..1841,1842..2633)
                     /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(1991..2010)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2804..3392
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3293..3312
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3546..3563
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3661..3795
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4220..4240)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4370..4435
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4862..5461)
                     /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(4945..4965)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5442..5461
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(join(5471..5500,1..300))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5485..5500,1..120)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
       61 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
      121 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
      181 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
      241 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
      301 attccacagc cggatccaag gtcgggcagg aagagggcct atttcccatg attccttcat
      361 atttgcatat acgatacaag gctgttagag agataattag aattaatttg actgtaaaca
      421 caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag
      481 ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg
      541 atttcttggc tttatatatc ttgtggaaag gacgaaacac cgtgctcgct tcggcagcac
      601 atatactagt cgacgggccg cactcgccgg tcccaagccc ggataaaatg ggagggggcg
      661 ggaaaccgcc taaccatgcc gagtgcggcc gcttgccatg tgctacttca acagtgcttg
      721 gacgctctga tgaataaccg aagtagttta tgggctaccg aaatcattca tggcaagtgg
      781 ccgcggtcgg cgtggactgt agaacactgc caatgccggt cccaagcccg gataaaagtg
      841 gagggtacag tccacgctct agagcgcact tcggtgcgct ttttactagg acctgcaggc
      901 atgcaagctt gacgtcggtt accgatatcc atatggcgac cgcatcgatc tcgagccgag
      961 gactagtaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     1021 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1081 tgtatcttat catgtcttac gtagataagt agcatggcgg gttatagcga agaggcccgc
     1141 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggaaatt gtaagcgtta
     1201 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     1261 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     1321 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     1381 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     1441 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     1501 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     1561 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     1621 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1681 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1741 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1801 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1861 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1921 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     1981 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     2041 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2101 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2161 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2221 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2281 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     2341 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2401 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2461 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2521 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2581 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2641 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     2701 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2761 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2821 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2881 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2941 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3001 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3061 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3121 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3181 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3241 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3301 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3361 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3421 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3481 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3541 agcgcagcga gtcagtgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
     3601 ttcctggagg ggtggagtcg tgacgtgaat tacgtcatag ggttagggag gtcctggatc
     3661 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     3721 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     3781 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     3841 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat
     3901 ctctagtcaa ggcactatac atcaaatatt ccttattaac ccctttacaa attaaaaagc
     3961 taaaggtaca caatttttga gcatagttat taatagcaga cactctatgc ctgtgtggag
     4021 taagaaaaaa cagtatgtta tgattataac tgttatgcct acttataaag gttacagaat
     4081 atttttccat aattttcttg tatagcagtg cagctttttc ctttgtggtg taaatagcaa
     4141 agcaagcaag agttctatta ctaaacacag catgactcaa aaaacttagc aattctgaag
     4201 gaaagtcctt ggggtcttct acctttctct tcttttttgg aggagtagaa tgttgagagt
     4261 cagcagtagc ctcatcatca ctagatggca tttcttctga gcaaaacagg ttttcctcat
     4321 taaaggcatt ccaccactgc tcccattcat cagttccata ggttggaatc taaaatacac
     4381 aaacaattag aatcagtagt ttaacacatt atacacttaa aaattttata tttaccttag
     4441 agctttaaat ctctgtaggt agtttgtcca attatgtcac accacagaag taaggttcct
     4501 tcacaaagat ccggaccaaa gcggccatcg tgcctcccca ctcctgcagt tcgggggcat
     4561 ggatgcgcgg atagccgctg ctggtttcct ggatgccgac ggatttgcac tgccggtaga
     4621 actccgcgag gtcgtccagc ctcaggcagc agctgaacca actcgcgagg ggatcgagcc
     4681 cggggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt
     4741 cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc
     4801 gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa ctgcgggtcg tggggcgggc
     4861 gtcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc acctcgacgt
     4921 cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc gcggaggtct
     4981 ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac tccggggagc acgacggcgc
     5041 tgcccagacc cttgccctgg tggtcgggcg agacgccgac ggtggccagg aaccacgcgg
     5101 gctccttggg ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg gccagccggg
     5161 aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa caccgccccc gcttcgacgc
     5221 tctccggcgt ggtccagacc gccacggcag caccgtcgtc cgcgacccac accttgccga
     5281 tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc tcgatgtggc
     5341 gatcaggatc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc
     5401 gtacggccct ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca
     5461 tggtaagctt tttgcaaaag cctaggcctc caaaaaagcc
//
