LOCUS       sequence-333870-        5496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of circular RNA (Broccoli-based SAM sensor).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5496)
  AUTHORS   Litke JL, Jaffrey SR
  TITLE     Highly efficient expression of circular RNA aptamers in cells using 
            autocatalytic transcripts.
  JOURNAL   Nat Biotechnol. 2019 Apr 8. pii: 10.1038/s41587-019-0090-6. doi: 
            10.1038/s41587-019-0090-6.
  PUBMED    30962542
REFERENCE   2  (bases 1 to 5496)
  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: "Nat 
            Biotechnol. 2019 Apr 8. pii: 10.1038/s41587-019-0090-6. doi: 
            10.1038/s41587-019-0090-6."
COMMENT     Sequence Label: pAV-U6+27-Tornado-BroccoliSamSensor#1
FEATURES             Location/Qualifiers
     source          1..5496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..273
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     25..45
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     196..215
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    656..777
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(874..1329)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1011..1032)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1223..1242
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1356..1460
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1461..1529,1530..2321)
                     /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(1679..1698)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2492..3080
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2981..3000
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3234..3251
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3349..3483
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3908..3928)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4058..4123
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4550..5149)
                     /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(4633..4653)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5130..5149
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(5159..5488)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5173..5308
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5473..5493
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 gccggatcca aggtcgggca ggaagagggc ctatttccca tgattccttc atatttgcat
       61 atacgataca aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata
      121 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
      181 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
      241 gctttatata tcttgtggaa aggacgaaac accgtgctcg cttcggcagc acatatacta
      301 gtcgacgggc cgcactcgcc ggtcccaagc ccggataaaa tgggaggggg cgggaaaccg
      361 cctaaccatg ccgagtgcgg ccgcgagacg gtcgggtcca cgaaaggatg gcggaaacgc
      421 cagatgcctt gtaaccgaaa gggtgtcgag tagagtgtgg gctcgtggcc gcggtcggcg
      481 tggactgtag aacactgcca atgccggtcc caagcccgga taaaagtgga gggtacagtc
      541 cacgctctag agcgcacttc ggtgcgcttt ttactaggac ctgcaggcat gcaagcttga
      601 cgtcggttac cgatatccat atggcgaccg catcgatctc gagccgagga ctagtaactt
      661 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
      721 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
      781 tgtcttacgt agataagtag catggcgggt tatagcgaag aggcccgcac cgatcgccct
      841 tcccaacagt tgcgcagcct gaatggcgaa tggaaattgt aagcgttaat attttgttaa
      901 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
      961 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     1021 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     1081 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     1141 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     1201 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     1261 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     1321 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1381 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1441 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1501 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1561 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1621 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1681 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     1741 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     1801 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     1861 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     1921 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     1981 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     2041 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     2101 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     2161 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     2221 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     2281 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2341 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2401 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2461 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2521 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2581 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     2641 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2701 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2761 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2821 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2881 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2941 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3001 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3061 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3121 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3181 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3241 cagtgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctggagggg
     3301 tggagtcgtg acgtgaatta cgtcataggg ttagggaggt cctggatcga tccagacatg
     3361 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     3421 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     3481 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     3541 ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatct ctagtcaagg
     3601 cactatacat caaatattcc ttattaaccc ctttacaaat taaaaagcta aaggtacaca
     3661 atttttgagc atagttatta atagcagaca ctctatgcct gtgtggagta agaaaaaaca
     3721 gtatgttatg attataactg ttatgcctac ttataaaggt tacagaatat ttttccataa
     3781 ttttcttgta tagcagtgca gctttttcct ttgtggtgta aatagcaaag caagcaagag
     3841 ttctattact aaacacagca tgactcaaaa aacttagcaa ttctgaagga aagtccttgg
     3901 ggtcttctac ctttctcttc ttttttggag gagtagaatg ttgagagtca gcagtagcct
     3961 catcatcact agatggcatt tcttctgagc aaaacaggtt ttcctcatta aaggcattcc
     4021 accactgctc ccattcatca gttccatagg ttggaatcta aaatacacaa acaattagaa
     4081 tcagtagttt aacacattat acacttaaaa attttatatt taccttagag ctttaaatct
     4141 ctgtaggtag tttgtccaat tatgtcacac cacagaagta aggttccttc acaaagatcc
     4201 ggaccaaagc ggccatcgtg cctccccact cctgcagttc gggggcatgg atgcgcggat
     4261 agccgctgct ggtttcctgg atgccgacgg atttgcactg ccggtagaac tccgcgaggt
     4321 cgtccagcct caggcagcag ctgaaccaac tcgcgagggg atcgagcccg gggtgggcga
     4381 agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga
     4441 ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt
     4501 tgggcgtcgc ttggtcggtc atttcgaact gcgggtcgtg gggcgggcgt caggcaccgg
     4561 gcttgcgggt catgcaccag gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg
     4621 tgaagccgag ccgctcgtag aaggggaggt tgcggggcgc ggaggtctcc aggaaggcgg
     4681 gcaccccggc gcgctcggcc gcctccactc cggggagcac gacggcgctg cccagaccct
     4741 tgccctggtg gtcgggcgag acgccgacgg tggccaggaa ccacgcgggc tccttgggcc
     4801 ggtgcggcgc caggaggcct tccatctgtt gctgcgcggc cagccgggaa ccgctcaact
     4861 cggccatgcg cgggccgatc tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg
     4921 tccagaccgc cacggcagca ccgtcgtccg cgacccacac cttgccgatg tcgagcccga
     4981 cgcgcgtgag gaagagttct tgcagctcgg tgacccgctc gatgtggcga tcaggatcga
     5041 cggtgtggcg cgtggcgggg tagtcggcga acgcggcggc gagggtgcgt acggccctgg
     5101 ggacgtcgtc gcgggtggcg aggcgcaccg tgggcttgta ctcggtcatg gtaagctttt
     5161 tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg
     5221 aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg
     5281 cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg
     5341 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     5401 acctggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     5461 gggactttcc acaccctaac tgacacacat tccaca
//
