LOCUS       sequence-430577-        5504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses acyclovir-controllable IRE variant #A4 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 5504)
  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 5504)
  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#A4
FEATURES             Location/Qualifiers
     source          1..5504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     224..244
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        272..520
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     272..292
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     443..462
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    911..1032
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1129..1584)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1266..1287)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1478..1497
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1611..1715
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1716..1784,1785..2576)
                     /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(1934..1953)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2747..3335
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3236..3255
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3489..3506
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3604..3738
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4163..4183)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4313..4378
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4805..5404)
                     /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(4888..4908)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5385..5404
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(join(5414..5504,1..239))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5428..5504,1..59)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg
       61 ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt
      121 ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg
      181 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca
      241 ttccacagcc ggatccaagg tcgggcagga agagggccta tttcccatga ttccttcata
      301 tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac
      361 aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt
      421 tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga
      481 tttcttggct ttatatatct tgtggaaagg acgaaacacc gtgctcgctt cggcagcaca
      541 tatactagtc gacgggccgc actcgccggt cccaagcccg gataaaatgg gagggggcgg
      601 gaaaccgcct aaccatgccg agtgcggccg cttgccatgt attatgctta aacagtgctt
      661 taactaattc tgatgaataa ccgaagtagt ttatgggcta ccgaaatcat tcatggcaag
      721 tggccgcggt cggcgtggac tgtagaacac tgccaatgcc ggtcccaagc ccggataaaa
      781 gtggagggta cagtccacgc tctagagcgc acttcggtgc gctttttact aggacctgca
      841 ggcatgcaag cttgacgtcg gttaccgata tccatatggc gaccgcatcg atctcgagcc
      901 gaggactagt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
      961 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     1021 caatgtatct tatcatgtct tacgtagata agtagcatgg cgggttatag cgaagaggcc
     1081 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggaa attgtaagcg
     1141 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     1201 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     1261 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     1321 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     1381 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     1441 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     1501 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     1561 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     1621 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     1681 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     1741 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     1801 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     1861 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     1921 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     1981 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     2041 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     2101 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     2161 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     2221 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     2281 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     2341 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     2401 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     2461 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     2521 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     2581 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     2641 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     2701 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2761 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2821 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2881 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2941 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3001 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3061 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3121 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3181 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3241 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3301 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3361 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3421 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     3481 ccgagcgcag cgagtcagtg agcgagcgcg cagagaggga gtggccaact ccatcactag
     3541 gggttcctgg aggggtggag tcgtgacgtg aattacgtca tagggttagg gaggtcctgg
     3601 atcgatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     3661 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3721 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3781 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat
     3841 gatctctagt caaggcacta tacatcaaat attccttatt aaccccttta caaattaaaa
     3901 agctaaaggt acacaatttt tgagcatagt tattaatagc agacactcta tgcctgtgtg
     3961 gagtaagaaa aaacagtatg ttatgattat aactgttatg cctacttata aaggttacag
     4021 aatatttttc cataattttc ttgtatagca gtgcagcttt ttcctttgtg gtgtaaatag
     4081 caaagcaagc aagagttcta ttactaaaca cagcatgact caaaaaactt agcaattctg
     4141 aaggaaagtc cttggggtct tctacctttc tcttcttttt tggaggagta gaatgttgag
     4201 agtcagcagt agcctcatca tcactagatg gcatttcttc tgagcaaaac aggttttcct
     4261 cattaaaggc attccaccac tgctcccatt catcagttcc ataggttgga atctaaaata
     4321 cacaaacaat tagaatcagt agtttaacac attatacact taaaaatttt atatttacct
     4381 tagagcttta aatctctgta ggtagtttgt ccaattatgt cacaccacag aagtaaggtt
     4441 ccttcacaaa gatccggacc aaagcggcca tcgtgcctcc ccactcctgc agttcggggg
     4501 catggatgcg cggatagccg ctgctggttt cctggatgcc gacggatttg cactgccggt
     4561 agaactccgc gaggtcgtcc agcctcaggc agcagctgaa ccaactcgcg aggggatcga
     4621 gcccggggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg
     4681 cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat
     4741 ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaactgcggg tcgtggggcg
     4801 ggcgtcaggc accgggcttg cgggtcatgc accaggtgcg cggtccttcg ggcacctcga
     4861 cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg ggcgcggagg
     4921 tctccaggaa ggcgggcacc ccggcgcgct cggccgcctc cactccgggg agcacgacgg
     4981 cgctgcccag acccttgccc tggtggtcgg gcgagacgcc gacggtggcc aggaaccacg
     5041 cgggctcctt gggccggtgc ggcgccagga ggccttccat ctgttgctgc gcggccagcc
     5101 gggaaccgct caactcggcc atgcgcgggc cgatctcggc gaacaccgcc cccgcttcga
     5161 cgctctccgg cgtggtccag accgccacgg cagcaccgtc gtccgcgacc cacaccttgc
     5221 cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc cgctcgatgt
     5281 ggcgatcagg atcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg gcggcgaggg
     5341 tgcgtacggc cctggggacg tcgtcgcggg tggcgaggcg caccgtgggc ttgtactcgg
     5401 tcatggtaag ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga
     5461 atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaa
//
