LOCUS       sequence-354582-        6224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Squash aptamer in mammalian cells using the U6+27 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6224)
  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 6224)
  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-Squash
FEATURES             Location/Qualifiers
     source          1..6224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     607..626
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1037..1366)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1051..1186
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1351..1371
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1399..1647
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1399..1419
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1570..1589
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    1874..1995
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   2044..2173
                     /label=AAV2 ITR
     rep_origin      complement(2267..2722)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2404..2425)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2616..2635
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2749..2853
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2854..2922,2923..3714)
                     /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(3072..3091)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3885..4473
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4374..4393
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4916..5050
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5475..5495)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          5625..5690
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(join(6110..6224,1..680))
                     /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(join(6221..6224,1..16))
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
       61 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
      121 gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct
      181 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
      241 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
      301 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
      361 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
      421 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
      481 agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
      541 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
      601 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
      661 aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agatcttgat
      721 cccctgcgcc atcagatcct tggcggcaag aaagccatcc agtttacttt gcagggcttc
      781 ccaaccttac cagagggcgc cccagctggc aattccggtt cgcttgctgt ccataaaacc
      841 gcccagtcta gctatcgcca tgtaagccca ctgcaagcta cctgctttct ctttgcgctt
      901 gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc
      961 tgcggactgg ctttctacgt gttccgcttc ctttagcagc ccttgcgccc tgagtgcttg
     1021 cggcagcgtg aagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct
     1081 ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc
     1141 catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg
     1201 gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg
     1261 agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc
     1321 tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc cacagccgga
     1381 tccaaggtcg ggcaggaaga gggcctattt cccatgattc cttcatattt gcatatacga
     1441 tacaaggctg ttagagagat aattagaatt aatttgactg taaacacaaa gatattagta
     1501 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
     1561 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta
     1621 tatatcttgt ggaaaggacg aaacaccgtg ctcgcttcgg cagcacatat actagtcgac
     1681 cctaggctac aaggtgagcc caataatacg gtttgggtta ggataggaag tagagccgta
     1741 aactctctaa gcggtagtct aggtctagag cggacttcgg tccgcttttt actaggacct
     1801 gcaggcatgc aagcttgacg tcggttaccg atatccatat ggcggccgca tcgatctcga
     1861 gccgcggact agtaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     1921 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     1981 catcaatgta tcttatcatg tcttacgtag ataagtagca tggcgggtta atcattaact
     2041 acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     2101 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     2161 agcgagcgcg cagagaggga cagatctgca gatctgctgg cgtaatagcg aagaggcccg
     2221 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggaaat tgtaagcgtt
     2281 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2341 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2401 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2461 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2521 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2581 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2641 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2701 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2761 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2821 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     2881 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     2941 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     3001 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     3061 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     3121 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     3181 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     3241 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     3301 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     3361 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     3421 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     3481 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3541 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3601 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3661 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     3721 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3781 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3841 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3901 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3961 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4021 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4081 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4141 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4201 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4261 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4321 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4381 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4441 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4501 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4561 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4621 gagcgcagcg agtcagtgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
     4681 gttcctggag gggtggagtc gtgacgtgaa ttacgtcata gggttaggga ggtcctggat
     4741 nncsrtmywr mcrcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     4801 agcgagtcag tgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
     4861 ggaggggtgg agtcgtgacg tgaattacgt catagggtta gggaggtcct ggatcgatcc
     4921 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     4981 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     5041 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     5101 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatctcta
     5161 gtcaaggcac tatacatcaa atattcctta ttaacccctt tacaaattaa aaagctaaag
     5221 gtacacaatt tttgagcata gttattaata gcagacactc tatgcctgtg tggagtaaga
     5281 aaaaacagta tgttatgatt ataactgtta tgcctactta taaaggttac agaatatttt
     5341 tccataattt tcttgtatag cagtgcagct ttttcctttg tggtgtaaat agcaaagcaa
     5401 gcaagagttc tattactaaa cacagcatga ctcaaaaaac ttagcaattc tgaaggaaag
     5461 tccttggggt cttctacctt tctcttcttt tttggaggag tagaatgttg agagtcagca
     5521 gtagcctcat catcactaga tggcatttct tctgagcaaa acaggttttc ctcattaaag
     5581 gcattccacc actgctccca ttcatcagtt ccataggttg gaatctaaaa tacacaaaca
     5641 attagaatca gtagtttaac acattataca cttaaaaatt ttatatttac cttagagctt
     5701 taaatctctg taggtagttt gtccaattat gtcacaccac agaagtaagg ttccttcaca
     5761 aagatccgga ccaaagcggc catcgtgcct ccccactcct gcagttcggg ggcatggatg
     5821 cgcggatagc cgctgctggt ttcctggatg ccgacggatt tgcactgccg gtagaactcc
     5881 gcgaggtcgt ccagcctcag gcagcagctg aaccaactcg cgaggggatc gagcccgggg
     5941 tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg
     6001 aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtgat
     6061 ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct cagaagaact
     6121 cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca
     6181 cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aata
//
