LOCUS       sequence-430361-        5529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses acyclovir-controllable Broccoli aptamer #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 5529)
  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 5529)
  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-U6+27-F30-Tornado-Acyclovir-Broccoli#4
FEATURES             Location/Qualifiers
     source          1..5529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(26..355)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      40..175
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     340..360
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        388..636
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     388..408
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     559..578
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    1052..1173
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1270..1725)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1407..1428)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1619..1638
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1752..1856
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1857..1925,1926..2717)
                     /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(2075..2094)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2888..3476
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3377..3396
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3630..3647
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3745..3879
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4304..4324)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4454..4519
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(join(4946..5529,1..16))
                     /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(5029..5049)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     join(5526..5529,1..16)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 gcttgtactc ggtcatggta agctttttgc aaaagcctag gcctccaaaa aagcctcctc
       61 actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa
      121 ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg
      181 gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc
      241 ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt
      301 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc
      361 acagccggat ccaaggtcgg gcaggaagag ggcctatttc ccatgattcc ttcatatttg
      421 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag
      481 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
      541 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
      601 ttggctttat atatcttgtg gaaaggacga aacaccgtgc tcgcttcggc agcacatata
      661 ctagtcgacg ggccgcactc gccggtccca agcccggata aaatgggagg gggcgggaaa
      721 ccgcctaacc atgccgagtg cggccgcggt tgccatgtgt tctgtcgagt agagtgtggg
      781 ctcttcggag acggtcgggt ccagaactct gatgaataac cgaagtagtt tatgggctac
      841 cgaaatcatt catggcaacc ggtgccgcgg tcggcgtgga ctgtagaaca ctgccaatgc
      901 cggtcccaag cccggataaa agtggagggt acagtccacg ctctagagcg cacttcggtg
      961 cgctttttac taggacctgc aggcatgcaa gcttgacgtc ggttaccgat atccatatgg
     1021 cgaccgcatc gatctcgagc cgaggactag taacttgttt attgcagctt ataatggtta
     1081 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     1141 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc ttacgtagat aagtagcatg
     1201 gcgggttata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     1261 ggcgaatgga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     1321 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     1381 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     1441 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     1501 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     1561 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     1621 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     1681 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     1741 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1801 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     1861 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1921 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     1981 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2041 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     2101 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2161 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2221 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2281 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2341 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2401 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2461 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2521 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     2581 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2641 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     2701 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     2761 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     2821 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     2881 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     2941 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3001 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3061 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3121 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3181 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3241 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3301 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3361 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3421 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3481 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     3541 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     3601 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgagcgc gcagagaggg
     3661 agtggccaac tccatcacta ggggttcctg gaggggtgga gtcgtgacgt gaattacgtc
     3721 atagggttag ggaggtcctg gatcgatcca gacatgataa gatacattga tgagtttgga
     3781 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     3841 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     3901 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     3961 aaatgtggta tggctgatta tgatctctag tcaaggcact atacatcaaa tattccttat
     4021 taaccccttt acaaattaaa aagctaaagg tacacaattt ttgagcatag ttattaatag
     4081 cagacactct atgcctgtgt ggagtaagaa aaaacagtat gttatgatta taactgttat
     4141 gcctacttat aaaggttaca gaatattttt ccataatttt cttgtatagc agtgcagctt
     4201 tttcctttgt ggtgtaaata gcaaagcaag caagagttct attactaaac acagcatgac
     4261 tcaaaaaact tagcaattct gaaggaaagt ccttggggtc ttctaccttt ctcttctttt
     4321 ttggaggagt agaatgttga gagtcagcag tagcctcatc atcactagat ggcatttctt
     4381 ctgagcaaaa caggttttcc tcattaaagg cattccacca ctgctcccat tcatcagttc
     4441 cataggttgg aatctaaaat acacaaacaa ttagaatcag tagtttaaca cattatacac
     4501 ttaaaaattt tatatttacc ttagagcttt aaatctctgt aggtagtttg tccaattatg
     4561 tcacaccaca gaagtaaggt tccttcacaa agatccggac caaagcggcc atcgtgcctc
     4621 cccactcctg cagttcgggg gcatggatgc gcggatagcc gctgctggtt tcctggatgc
     4681 cgacggattt gcactgccgg tagaactccg cgaggtcgtc cagcctcagg cagcagctga
     4741 accaactcgc gaggggatcg agcccggggt gggcgaagaa ctccagcatg agatccccgc
     4801 gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga
     4861 aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt
     4921 cgaactgcgg gtcgtggggc gggcgtcagg caccgggctt gcgggtcatg caccaggtgc
     4981 gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc tcgtagaagg
     5041 ggaggttgcg gggcgcggag gtctccagga aggcgggcac cccggcgcgc tcggccgcct
     5101 ccactccggg gagcacgacg gcgctgccca gacccttgcc ctggtggtcg ggcgagacgc
     5161 cgacggtggc caggaaccac gcgggctcct tgggccggtg cggcgccagg aggccttcca
     5221 tctgttgctg cgcggccagc cgggaaccgc tcaactcggc catgcgcggg ccgatctcgg
     5281 cgaacaccgc ccccgcttcg acgctctccg gcgtggtcca gaccgccacg gcagcaccgt
     5341 cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag agttcttgca
     5401 gctcggtgac ccgctcgatg tggcgatcag gatcgacggt gtggcgcgtg gcggggtagt
     5461 cggcgaacgc ggcggcgagg gtgcgtacgg ccctggggac gtcgtcgcgg gtggcgaggc
     5521 gcaccgtgg
//
