LOCUS       sequence-354607-        6236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Squash-SAM sensor 4-2 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 6236)
  AUTHORS   Dey SK, Filonov GS, Olarerin-George AO, Jackson BT, Finley LWS, 
            Jaffrey SR
  TITLE     Repurposing an adenine riboswitch into a fluorogenic imaging and 
            sensing tag.
  JOURNAL   Nat Chem Biol. 2021 Dec 22. pii: 10.1038/s41589-021-00925-0. doi: 
            10.1038/s41589-021-00925-0.
  PUBMED    34937909
REFERENCE   2  (bases 1 to 6236)
  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 Chem 
            Biol. 2021 Dec 22. pii: 10.1038/s41589-021-00925-0. doi: 
            10.1038/s41589-021-00925-0."
COMMENT     Sequence Label: pAV-U6+27-Tornado-Squash-SAM-sensor 4-2
FEATURES             Location/Qualifiers
     source          1..6236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     70..90
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        118..366
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     118..138
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     289..308
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    779..900
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   949..1078
                     /label=AAV2 ITR
     rep_origin      complement(1172..1627)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1309..1330)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1521..1540
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1654..1758
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1759..1827,1828..2619)
                     /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(1977..1996)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2790..3378
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3279..3298
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3647..3781
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4206..4226)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4356..4421
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4841..5635)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4952..4971)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5562..5581
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5992..6236,1..85))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6006..6141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
       61 actttccaca ccctaactga cacacattcc acagccggat ccaaggtcgg gcaggaagag
      121 ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata
      181 attagaatta atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag
      241 taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg
      301 cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
      361 aacaccgtgc tcgcttcggc agcacatata ctagtcgacg ggccgcactc gccggtccca
      421 agcccggata aaatgggagg gggcgggaaa ccgcctaacc atgccgagtg cggccgccgc
      481 cagatgcctt gtaaccgaaa gggacacaag gtgagcccaa taatacggtt tgggttagga
      541 taggaagtag agccgtaaac tctctaagcg gtgtcgaaag gatggcggtg gccgcggtcg
      601 gcgtggactg tagaacactg ccaatgccgg tcccaagccc ggataaaagt ggagggtaca
      661 gtccacgctc tagagcggac ttcggtccgc tttttactag gacctgcagg catgcaagct
      721 tgacgtcggt taccgatatc catatggcga ccgcatcgat ctcgagccga ggactagtaa
      781 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
      841 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
      901 tcatgtctta cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag
      961 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     1021 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagag
     1081 agggacagat ctgcagatct gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     1141 ccaacagttg cgcagcctga atggcgaatg gaaattgtaa gcgttaatat tttgttaaaa
     1201 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     1261 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     1321 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     1381 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     1441 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     1501 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     1561 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     1621 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1681 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1741 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1801 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1861 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1921 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1981 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     2041 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     2101 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     2161 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2221 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2281 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     2341 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2401 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2461 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2521 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2581 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2641 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2701 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2761 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2821 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2881 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2941 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3001 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3061 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3121 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3181 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3241 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3301 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3361 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3421 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3481 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3541 gtgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc tggaggggtg
     3601 gagtcgtgac gtgaattacg tcatagggtt agggaggtcc tggatcgatc cagacatgat
     3661 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     3721 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3781 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     3841 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatctct agtcaaggca
     3901 ctatacatca aatattcctt attaacccct ttacaaatta aaaagctaaa ggtacacaat
     3961 ttttgagcat agttattaat agcagacact ctatgcctgt gtggagtaag aaaaaacagt
     4021 atgttatgat tataactgtt atgcctactt ataaaggtta cagaatattt ttccataatt
     4081 ttcttgtata gcagtgcagc tttttccttt gtggtgtaaa tagcaaagca agcaagagtt
     4141 ctattactaa acacagcatg actcaaaaaa cttagcaatt ctgaaggaaa gtccttgggg
     4201 tcttctacct ttctcttctt ttttggagga gtagaatgtt gagagtcagc agtagcctca
     4261 tcatcactag atggcatttc ttctgagcaa aacaggtttt cctcattaaa ggcattccac
     4321 cactgctccc attcatcagt tccataggtt ggaatctaaa atacacaaac aattagaatc
     4381 agtagtttaa cacattatac acttaaaaat tttatattta ccttagagct ttaaatctct
     4441 gtaggtagtt tgtccaatta tgtcacacca cagaagtaag gttccttcac aaagatccgg
     4501 accaaagcgg ccatcgtgcc tccccactcc tgcagttcgg gggcatggat gcgcggatag
     4561 ccgctgctgg tttcctggat gccgacggat ttgcactgcc ggtagaactc cgcgaggtcg
     4621 tccagcctca ggcagcagct gaaccaactc gcgaggggat cgagcccggg gtgggcgaag
     4681 aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt
     4741 ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg
     4801 ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa
     4861 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
     4921 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
     4981 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
     5041 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg
     5101 gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
     5161 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat
     5221 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
     5281 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc
     5341 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag
     5401 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt
     5461 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
     5521 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
     5581 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa
     5641 acgatcctca tcctgtctct tgatcagatc ttgatcccct gcgccatcag atccttggcg
     5701 gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag ggcgccccag
     5761 ctggcaattc cggttcgctt gctgtccata aaaccgccca gtctagctat cgccatgtaa
     5821 gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc cagatagccc
     5881 agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtgttcc
     5941 gcttccttta gcagcccttg cgccctgagt gcttgcggca gcgtgaagct ttttgcaaaa
     6001 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     6061 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     6121 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     6181 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacc
//
