LOCUS       sequence-384488-        6258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of circular biSiRA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6258)
  AUTHORS   Buhler B, Schokolowski J, Benderoth A, Englert D, Grun F, Jaschke A,
            Sunbul M
  TITLE     Avidity-based bright and photostable light-up aptamers for 
            single-molecule mRNA imaging.
  JOURNAL   Nat Chem Biol. 2023 Apr;19(4):478-487. doi: 
            10.1038/s41589-022-01228-8. Epub 2023 Jan 19.
  PUBMED    36658339
REFERENCE   2  (bases 1 to 6258)
  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"; doi: 
            "10.1038/s41589-022-01228-8"; journalName: "Nat Chem Biol"; date: 
            "2023-04"; volume: "19"; issue: "4"; pages: "478-487"
COMMENT     Sequence Label: pAV-U6+27-Tornado-biSiRA
FEATURES             Location/Qualifiers
     source          1..6258
                     /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    708..829
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   878..1007
                     /label=AAV2 ITR
     rep_origin      complement(1101..1556)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1238..1259)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1450..1469
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1583..1687
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1688..1756,1757..2548)
                     /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(1906..1925)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2719..3307
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3208..3227
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3576..3710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4135..4155)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4285..4350
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4770..5564)
                     /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(4881..4900)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5491..5510
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5921..6250)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5935..6070
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6235..6255
                     /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 ccgctgtggg aggccaccgg gtttgaaaac ctggctgctt
      421 cggcagttgt atcctttggc caggaggaga aatcctccac cccaccgggt ttgaaaacct
      481 ggctgcttcg gcagttgtat cctttgggga cccacagtgg ccgcggtcgg cgtggactgt
      541 agaacactgc caatgccggt cccaagcccg gataaaagtg gagggtacag tccacgctct
      601 agagcggact tcggtccgct ttttactagg acctgcaggc atgcaagctt gacgtcggtt
      661 accgatatcc atatggcgac cgcatcgatc tcgagccgag gactagtaac ttgtttattg
      721 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
      781 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtcttac
      841 gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg
      901 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
      961 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagaga gggacagatc
     1021 tgcagatctg ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     1081 gcagcctgaa tggcgaatgg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     1141 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     1201 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     1261 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     1321 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     1381 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     1441 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     1501 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     1561 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1621 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1681 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1741 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1801 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1861 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1921 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1981 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2041 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     2101 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     2161 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     2221 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2281 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2341 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2401 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2461 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2521 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2581 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2641 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2701 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2761 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2821 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2881 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2941 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3001 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3061 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3121 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3181 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3241 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3301 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3361 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3421 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagcg
     3481 cgcagagagg gagtggccaa ctccatcact aggggttcct ggaggggtgg agtcgtgacg
     3541 tgaattacgt catagggtta gggaggtcct ggatcgatcc agacatgata agatacattg
     3601 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     3661 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     3721 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     3781 aaaacctcta caaatgtggt atggctgatt atgatctcta gtcaaggcac tatacatcaa
     3841 atattcctta ttaacccctt tacaaattaa aaagctaaag gtacacaatt tttgagcata
     3901 gttattaata gcagacactc tatgcctgtg tggagtaaga aaaaacagta tgttatgatt
     3961 ataactgtta tgcctactta taaaggttac agaatatttt tccataattt tcttgtatag
     4021 cagtgcagct ttttcctttg tggtgtaaat agcaaagcaa gcaagagttc tattactaaa
     4081 cacagcatga ctcaaaaaac ttagcaattc tgaaggaaag tccttggggt cttctacctt
     4141 tctcttcttt tttggaggag tagaatgttg agagtcagca gtagcctcat catcactaga
     4201 tggcatttct tctgagcaaa acaggttttc ctcattaaag gcattccacc actgctccca
     4261 ttcatcagtt ccataggttg gaatctaaaa tacacaaaca attagaatca gtagtttaac
     4321 acattataca cttaaaaatt ttatatttac cttagagctt taaatctctg taggtagttt
     4381 gtccaattat gtcacaccac agaagtaagg ttccttcaca aagatccgga ccaaagcggc
     4441 catcgtgcct ccccactcct gcagttcggg ggcatggatg cgcggatagc cgctgctggt
     4501 ttcctggatg ccgacggatt tgcactgccg gtagaactcc gcgaggtcgt ccagcctcag
     4561 gcagcagctg aaccaactcg cgaggggatc gagcccgggg tgggcgaaga actccagcat
     4621 gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa
     4681 cctttcatag aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg
     4741 gtcggtcatt tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag
     4801 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
     4861 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc
     4921 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
     4981 ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgcgcgcc
     5041 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc
     5101 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg
     5161 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
     5221 atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg
     5281 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga
     5341 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca
     5401 ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg
     5461 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
     5521 caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat
     5581 cctgtctctt gatcagatct tgatcccctg cgccatcaga tccttggcgg caagaaagcc
     5641 atccagttta ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc
     5701 ggttcgcttg ctgtccataa aaccgcccag tctagctatc gccatgtaag cccactgcaa
     5761 gctacctgct ttctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca
     5821 ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtgttccg cttcctttag
     5881 cagcccttgc gccctgagtg cttgcggcag cgtgaagctt tttgcaaaag cctaggcctc
     5941 caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg
     6001 cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta
     6061 ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc
     6121 tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat
     6181 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta
     6241 actgacacac attccaca
//
