LOCUS       sequence-381799-        8051 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insert: intronic Tau authentic Alu and repeat elements. Introns are 
            flanked by the tau cDNA exons 10-12 with FTLD-Tau K317M mutant. 
            Expresses tau circular RNA 12-->10 KM with 3X flag tag in exon 10..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8051)
  AUTHORS   Welden JR, Margvelani G, Arizaca Maquera KA, Gudlavalleti B, Miranda
            Sardon SC, Campos AR, Robil N, Lee DC, Hernandez AG, Wang WX, Di J, 
            de la Grange P, Nelson PT, Stamm S
  TITLE     RNA editing of microtubule-associated protein tau circular RNAs 
            promotes their translation and tau tangle formation.
  JOURNAL   Nucleic Acids Res. 2022 Dec 9;50(22):12979-12996. doi: 
            10.1093/nar/gkac1129.
  PUBMED    36533443
REFERENCE   2  (bases 1 to 8051)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2022-12-9- 9"; volume: "50"; issue: 
            "22"; pages: "12979-12996"
COMMENT     Sequence Label: pcDNA3.1_Tau Authentic Alu 10-12K317M
FEATURES             Location/Qualifiers
     source          1..8051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2302..2367
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3249..3260
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(3642..3659)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3648..3872
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3918..4346
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4005..4024)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4215..4236
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4355..4375)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4360..4689
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4540..4675
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4756..5550
                     /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(4810..4829)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5420..5439
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5724..5845
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5761..5780)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5815..5834
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5894..5910)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5894..5910)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5907..5929)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5918..5934
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5942..5948,5949..5966,5967..5972))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5987..6008
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6125..6142)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6296..6884)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6376..6395)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7055..7846,7847..7915))
                     /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     7678..7697
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7916..8020)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattccca
      961 ggctgatggg gcctccaggg cagtggtttc cacccagcat tctcaaaaga gccactgagg
     1021 tctcagtgcc attttcaaga tttcggaagc ggcctgggca cggctggtcc ttcactggga
     1081 tcaccacttg gcaattattt acacctgaga cgaataaaaa ccagagtgct gagattacag
     1141 gcatggtggc ttacgcttgt aatcggcttt gggaagccga ggtgggctga ttgcttgagc
     1201 ccaggagttt caaactatcc tggacaacat agcatgacct cgtctctaca aaaaatacaa
     1261 aaaatttgcc aggtgtggtg gcatgtgcct gtggtcccag ctacttggga ggctgaagta
     1321 ggagaatccc ctgagccctg ggaagtcgag gctgcactga gccgtgatga tgtcactgca
     1381 ctccagcctg ggtgacaaag tgagacccta tctcacaaag aaaaaaaaca aaacaaaaaa
     1441 cccaaagcac actgtttcca ctgtttccag agttcctgag aggaaaggtc accgggtgag
     1501 gaagacgttc tcactgatct ggcagagaaa atgtccagtt tttccaactc cctaaaccat
     1561 ggttttctat ttcacagccc ccccttgcag ggacagagcc ctcgtacaga agggacaccc
     1621 cacatttgtc ttccccacaa agcggcctgt gtcctgccta cggggtcagg gcttctcaaa
     1681 cctggctgtg tgtcagaatc accaggggaa cttttcaaaa ctagagagac tgaagccaga
     1741 ctcctagatt ctaattctag gtcagggcta ggggctgaga ttgtaaaaat ccacaggtga
     1801 ttctgatgcc cggcaggctt gagaacagcc gcagggagtt ctctgggaat gtgccggtgg
     1861 gtctagccag gtgtgagtgg agatgccggg gaacttccta ttactcactc gtcagtgtgg
     1921 ccgaacacat ttttcacttg acctcaggct ggtgaacgct cccctctggg gttcaggcct
     1981 cacgatgcca tccttttgtg aagtgaggac ctgcaatccc agcttcgtaa agcccgctgg
     2041 aaatcactca cacttctggg atgccttcag agcagccctc tatcccttca gctcccctgg
     2101 gatgtgactc gacctcccgt cactccccag actgcctctg ccaagtccga aagtggaggc
     2161 atccttgcga gcaagtaggc gggtccaggg tggcgcatgt cactcatcga aagtggaggc
     2221 gtccttgcga gcaagcaggc gggtccaggg tggcgtgtca ctcatccttt tttctggcta
     2281 ccaaaggtgc agataattaa tgactacaaa gaccatgacg gtgattataa agatcatgac
     2341 atcgattaca aggatgacga tgacaagaag aagctggatc ttagcaacgt ccagtccaag
     2401 tgtggctcaa aggataatat caaacacgtc ccgggaggcg gcagtgtgca aatagtctac
     2461 aaaccagttg acctgagcat ggtgacctcc aagtgtggct cattaggcaa catccatcat
     2521 aaaccaggag gtggccaggt ggaagtaaaa tctgagaagc ttgacttcaa ggacagagtc
     2581 cagtcgaaga ttgggtccct ggacaatatc acccacgtcc ctggcggagg aaataaaaag
     2641 gtaaaggggg tagggtgggt tggatgctgc ccttgggtat atgggcatta atcaagttga
     2701 gtggacaaag gctggtccag ttcccagagg aggaaaacag aggcttctgt gttgactggc
     2761 tggatgtggg ccctcagcag catccagtgg gtctccactg cctgtctcaa tcacctggag
     2821 ctttagcacg tttcacacct gggccccaac ctggagaggc tgaccaatgg gtctcagggg
     2881 cagctcggtt gctggagttt ttgtttttat ttatttttat gtatttaagg cagggtctct
     2941 gtattagtcc attctcacac tgctaataaa gacataccca agactgggta atttataaag
     3001 gaaagaggtt taatggactc acagttccat atggctgggg aggcctcaaa atcatggcgg
     3061 aaggcaaagg agaagcaaag gcatgtctta catggcaaca ggcaagagag cgtgtgcagg
     3121 ggaactccca tttataaaac catcagacct catgagattt attcactatc atgagaacag
     3181 catgggaaag acccgccccc atgattcagt tacctcccac tgggtccctc ccatgacaca
     3241 tggaattatg ggagctacaa ttcaagatga gatttgggtg gggacacagc caaaccatat
     3301 cagtctccct ctgtcatcca ggctggagtg cactggcatg atctcggctc actgcagcct
     3361 ctacctccct gggtcaggtg atcttcccac ctcagcctcc caggtagctg gaactacagg
     3421 tacctgccac tatgcctggc taaatatttt gtatttcctg tggagacgag gttttgccac
     3481 gttgcccagg ctggtcttga actcctgagg tcaagcaata tgcccacctc ggcctcccaa
     3541 ggtgctggga ttacaggtgt gagccacagt gctcggccta agtcactgca gtttttagcg
     3601 gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     3661 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3721 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3781 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     3841 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     3901 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3961 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4021 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4081 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4141 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4201 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4261 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4321 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4381 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4441 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4501 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4561 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4621 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4681 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     4741 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4801 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4861 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4921 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4981 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5041 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5101 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5161 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5221 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5281 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5341 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5401 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5461 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5521 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5581 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5641 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5701 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     5761 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5821 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5881 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5941 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6001 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6061 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6121 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6181 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6241 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6301 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6361 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6421 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6481 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6541 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6601 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6661 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6721 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6781 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6841 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6901 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6961 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     7021 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7081 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7141 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     7201 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7261 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7321 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7381 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7441 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7501 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7561 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7621 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7681 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7741 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7801 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7861 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7921 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7981 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     8041 cacctgacgt c
//
