LOCUS       sequence-430363-        5500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses acyclovir-controllable IRE variant #A8-3 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#A8-3
FEATURES             Location/Qualifiers
     source          1..5500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     210..230
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        258..506
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     258..278
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     429..448
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    893..1014
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1111..1566)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1248..1269)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1460..1479
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1593..1697
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1698..1766,1767..2558)
                     /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(1916..1935)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2729..3317
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3218..3237
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3471..3488
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3586..3720
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4145..4165)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4295..4360
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4787..5386)
                     /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(4870..4890)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5367..5386
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(join(5396..5500,1..225))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5410..5500,1..45)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
       61 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
      121 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct
      181 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagccggat
      241 ccaaggtcgg gcaggaagag ggcctatttc ccatgattcc ttcatatttg catatacgat
      301 acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag atattagtac
      361 aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt
      421 ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat
      481 atatcttgtg gaaaggacga aacaccgtgc tcgcttcggc agcacatata ctagtcgacg
      541 ggccgcactc gccggtccca agcccggata aaatgggagg gggcgggaaa ccgcctaacc
      601 atgccgagtg cggccgcttg ccatgtgctg gttcaacagt gcttggacgc tctgatgaat
      661 aaccgaagta gtttatgggc taccgaaatc attcatggca agtggccgcg gtcggcgtgg
      721 actgtagaac actgccaatg ccggtcccaa gcccggataa aagtggaggg tacagtccac
      781 gctctagagc gcacttcggt gcgcttttta ctaggacctg caggcatgca agcttgacgt
      841 cggttaccga tatccatatg gcgaccgcat cgatctcgag ccgaggacta gtaacttgtt
      901 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
      961 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     1021 cttacgtaga taagtagcat ggcgggttat agcgaagagg cccgcaccga tcgcccttcc
     1081 caacagttgc gcagcctgaa tggcgaatgg aaattgtaag cgttaatatt ttgttaaaat
     1141 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1201 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1261 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1321 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1381 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1441 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1501 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1561 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1621 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1681 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1741 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     1801 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     1861 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     1921 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     1981 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2041 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2101 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2161 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2221 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2281 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2341 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2401 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2461 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2521 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2581 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     2641 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2701 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2761 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2821 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     2881 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2941 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3001 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3061 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3121 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3181 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3241 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3301 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3361 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3421 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3481 tgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct ggaggggtgg
     3541 agtcgtgacg tgaattacgt catagggtta gggaggtcct ggatcgatcc agacatgata
     3601 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     3661 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     3721 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     3781 taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatctcta gtcaaggcac
     3841 tatacatcaa atattcctta ttaacccctt tacaaattaa aaagctaaag gtacacaatt
     3901 tttgagcata gttattaata gcagacactc tatgcctgtg tggagtaaga aaaaacagta
     3961 tgttatgatt ataactgtta tgcctactta taaaggttac agaatatttt tccataattt
     4021 tcttgtatag cagtgcagct ttttcctttg tggtgtaaat agcaaagcaa gcaagagttc
     4081 tattactaaa cacagcatga ctcaaaaaac ttagcaattc tgaaggaaag tccttggggt
     4141 cttctacctt tctcttcttt tttggaggag tagaatgttg agagtcagca gtagcctcat
     4201 catcactaga tggcatttct tctgagcaaa acaggttttc ctcattaaag gcattccacc
     4261 actgctccca ttcatcagtt ccataggttg gaatctaaaa tacacaaaca attagaatca
     4321 gtagtttaac acattataca cttaaaaatt ttatatttac cttagagctt taaatctctg
     4381 taggtagttt gtccaattat gtcacaccac agaagtaagg ttccttcaca aagatccgga
     4441 ccaaagcggc catcgtgcct ccccactcct gcagttcggg ggcatggatg cgcggatagc
     4501 cgctgctggt ttcctggatg ccgacggatt tgcactgccg gtagaactcc gcgaggtcgt
     4561 ccagcctcag gcagcagctg aaccaactcg cgaggggatc gagcccgggg tgggcgaaga
     4621 actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc
     4681 cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg
     4741 gcgtcgcttg gtcggtcatt tcgaactgcg ggtcgtgggg cgggcgtcag gcaccgggct
     4801 tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg gtgacggtga
     4861 agccgagccg ctcgtagaag gggaggttgc ggggcgcgga ggtctccagg aaggcgggca
     4921 ccccggcgcg ctcggccgcc tccactccgg ggagcacgac ggcgctgccc agacccttgc
     4981 cctggtggtc gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc ttgggccggt
     5041 gcggcgccag gaggccttcc atctgttgct gcgcggccag ccgggaaccg ctcaactcgg
     5101 ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc gacgctctcc ggcgtggtcc
     5161 agaccgccac ggcagcaccg tcgtccgcga cccacacctt gccgatgtcg agcccgacgc
     5221 gcgtgaggaa gagttcttgc agctcggtga cccgctcgat gtggcgatca ggatcgacgg
     5281 tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg gccctgggga
     5341 cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc ggtcatggta agctttttgc
     5401 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg
     5461 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
//
