LOCUS       sequence-430360-        5500 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 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
FEATURES             Location/Qualifiers
     source          1..5500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..112
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        140..388
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     140..160
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     311..330
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    775..896
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(993..1448)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1130..1151)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1342..1361
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1475..1579
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1580..1648,1649..2440)
                     /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(1798..1817)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2611..3199
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3100..3119
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3353..3370
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3468..3602
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4027..4047)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4177..4242
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4669..5268)
                     /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(4752..4772)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5249..5268
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(join(5278..5500,1..107))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5292..5427
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt
       61 ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt ccacagccgg
      121 atccaaggtc gggcaggaag agggcctatt tcccatgatt ccttcatatt tgcatatacg
      181 atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa agatattagt
      241 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
      301 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
      361 atatatcttg tggaaaggac gaaacaccgt gctcgcttcg gcagcacata tactagtcga
      421 cgggccgcac tcgccggtcc caagcccgga taaaatggga gggggcggga aaccgcctaa
      481 ccatgccgag tgcggccgct tgccatgtgc tgcttcaaca gtgcttggac gctctgatga
      541 ataaccgaag tagtttatgg gctaccgaaa tcattcatgg caagtggccg cggtcggcgt
      601 ggactgtaga acactgccaa tgccggtccc aagcccggat aaaagtggag ggtacagtcc
      661 acgctctaga gcgcacttcg gtgcgctttt tactaggacc tgcaggcatg caagcttgac
      721 gtcggttacc gatatccata tggcgaccgc atcgatctcg agccgaggac tagtaacttg
      781 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
      841 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
      901 gtcttacgta gataagtagc atggcgggtt atagcgaaga ggcccgcacc gatcgccctt
      961 cccaacagtt gcgcagcctg aatggcgaat ggaaattgta agcgttaata ttttgttaaa
     1021 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     1081 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     1141 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     1201 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     1261 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     1321 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     1381 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     1441 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     1501 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     1561 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     1621 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     1681 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     1741 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     1801 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     1861 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     1921 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     1981 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     2041 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     2101 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     2161 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     2221 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     2281 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     2341 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     2401 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     2461 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     2521 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2581 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2641 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2701 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2761 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2821 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2881 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2941 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3001 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3061 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3121 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3181 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3241 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3301 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3361 agtgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc ctggaggggt
     3421 ggagtcgtga cgtgaattac gtcatagggt tagggaggtc ctggatcgat ccagacatga
     3481 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     3541 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     3601 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     3661 tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatctc tagtcaaggc
     3721 actatacatc aaatattcct tattaacccc tttacaaatt aaaaagctaa aggtacacaa
     3781 tttttgagca tagttattaa tagcagacac tctatgcctg tgtggagtaa gaaaaaacag
     3841 tatgttatga ttataactgt tatgcctact tataaaggtt acagaatatt tttccataat
     3901 tttcttgtat agcagtgcag ctttttcctt tgtggtgtaa atagcaaagc aagcaagagt
     3961 tctattacta aacacagcat gactcaaaaa acttagcaat tctgaaggaa agtccttggg
     4021 gtcttctacc tttctcttct tttttggagg agtagaatgt tgagagtcag cagtagcctc
     4081 atcatcacta gatggcattt cttctgagca aaacaggttt tcctcattaa aggcattcca
     4141 ccactgctcc cattcatcag ttccataggt tggaatctaa aatacacaaa caattagaat
     4201 cagtagttta acacattata cacttaaaaa ttttatattt accttagagc tttaaatctc
     4261 tgtaggtagt ttgtccaatt atgtcacacc acagaagtaa ggttccttca caaagatccg
     4321 gaccaaagcg gccatcgtgc ctccccactc ctgcagttcg ggggcatgga tgcgcggata
     4381 gccgctgctg gtttcctgga tgccgacgga tttgcactgc cggtagaact ccgcgaggtc
     4441 gtccagcctc aggcagcagc tgaaccaact cgcgagggga tcgagcccgg ggtgggcgaa
     4501 gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat
     4561 tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt
     4621 gggcgtcgct tggtcggtca tttcgaactg cgggtcgtgg ggcgggcgtc aggcaccggg
     4681 cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt
     4741 gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg
     4801 caccccggcg cgctcggccg cctccactcc ggggagcacg acggcgctgc ccagaccctt
     4861 gccctggtgg tcgggcgaga cgccgacggt ggccaggaac cacgcgggct ccttgggccg
     4921 gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc agccgggaac cgctcaactc
     4981 ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt
     5041 ccagaccgcc acggcagcac cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac
     5101 gcgcgtgagg aagagttctt gcagctcggt gacccgctcg atgtggcgat caggatcgac
     5161 ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggccctggg
     5221 gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg taagcttttt
     5281 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     5341 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     5401 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     5461 ctaattgaga tgcatgcttt gcatacttct gcctgctggg
//
