LOCUS       sequence-354583-        5968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 5S rRNA fused to Squash aptamer in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5968)
  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 5968)
  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-5S-Squash
FEATURES             Location/Qualifiers
     source          1..5968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    93..227
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(652..672)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          802..867
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(1287..2081)
                     /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(1398..1417)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2008..2027
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2438..2767)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2452..2587
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2752..2772
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    3189..3310
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   3359..3488
                     /label=AAV2 ITR
     rep_origin      complement(3582..4037)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3719..3740)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3931..3950
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4064..4168
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4169..4237,4238..5029)
                     /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(4387..4406)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5200..5788
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5689..5708
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agagagggag tggccaactc catcactagg ggttcctgga ggggtggagt cgtgacgtga
       61 attacgtcat agggttaggg aggtcctgga tcgatccaga catgataaga tacattgatg
      121 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
      181 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
      241 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
      301 acctctacaa atgtggtatg gctgattatg atctctagtc aaggcactat acatcaaata
      361 ttccttatta acccctttac aaattaaaaa gctaaaggta cacaattttt gagcatagtt
      421 attaatagca gacactctat gcctgtgtgg agtaagaaaa aacagtatgt tatgattata
      481 actgttatgc ctacttataa aggttacaga atatttttcc ataattttct tgtatagcag
      541 tgcagctttt tcctttgtgg tgtaaatagc aaagcaagca agagttctat tactaaacac
      601 agcatgactc aaaaaactta gcaattctga aggaaagtcc ttggggtctt ctacctttct
      661 cttctttttt ggaggagtag aatgttgaga gtcagcagta gcctcatcat cactagatgg
      721 catttcttct gagcaaaaca ggttttcctc attaaaggca ttccaccact gctcccattc
      781 atcagttcca taggttggaa tctaaaatac acaaacaatt agaatcagta gtttaacaca
      841 ttatacactt aaaaatttta tatttacctt agagctttaa atctctgtag gtagtttgtc
      901 caattatgtc acaccacaga agtaaggttc cttcacaaag atccggacca aagcggccat
      961 cgtgcctccc cactcctgca gttcgggggc atggatgcgc ggatagccgc tgctggtttc
     1021 ctggatgccg acggatttgc actgccggta gaactccgcg aggtcgtcca gcctcaggca
     1081 gcagctgaac caactcgcga ggggatcgag cccggggtgg gcgaagaact ccagcatgag
     1141 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
     1201 ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc
     1261 ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg
     1321 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
     1381 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
     1441 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
     1501 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg
     1561 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     1621 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     1681 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     1741 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     1801 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     1861 cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg
     1921 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     1981 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     2041 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct
     2101 gtctcttgat cagatcttga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc
     2161 cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggt
     2221 tcgcttgctg tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct
     2281 acctgctttc tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc
     2341 atccggggtc agcaccgttt ctgcggactg gctttctacg tgttccgctt cctttagcag
     2401 cccttgcgcc ctgagtgctt gcggcagcgt gaagcttttt gcaaaagcct aggcctccaa
     2461 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
     2521 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
     2581 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt
     2641 gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga
     2701 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact
     2761 gacacacatt ccacagccgg atccggcccc gggctggcgg tgtcggatgc aatccggcgg
     2821 gcacggccgg gccgggctgg gctcttgggg cagccaggcg cctccttcag cgtctacggc
     2881 cataccaccc tgaacgcgcc cgatctcgtc tgatctcgga agctaagcag ggtcgggcct
     2941 ggttagtact tggatgggag accgcctggg aataccgggt gctgtaggcg tcgaccctag
     3001 gctacaaggt gagcccaata atacggtttg ggttaggata ggaagtagag ccgtaaactc
     3061 tctaagcggt agtctaggtc tagagcggac ttcggtccgc tttttactag gacctgcagg
     3121 catgcaagct tgacgtcggt taccgatatc catatggcgg ccgcatcgat ctcgagccgc
     3181 ggactagtaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3241 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3301 atgtatctta tcatgtctta cgtagataag tagcatggcg ggttaatcat taactacaag
     3361 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     3421 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     3481 gcgcgcagag agggacagat ctgcagatct gctggcgtaa tagcgaagag gcccgcaccg
     3541 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gaaattgtaa gcgttaatat
     3601 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3661 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3721 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3781 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     3841 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3901 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3961 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4021 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4081 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4141 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4201 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4261 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4321 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4381 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4441 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4501 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4561 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4621 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4681 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4741 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4801 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4861 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4921 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4981 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5041 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5101 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtaagt tttcgttcca
     5161 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5221 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5281 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5341 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5401 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5461 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5521 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5581 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5641 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5701 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5761 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5821 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5881 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     5941 cagcgagtca gtgagcgagc gagcgcgc
//
