LOCUS       sequence-381810-        5726 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insert contains linear tau 0N4R with a 3X Flag tag. Expresses linear
            protein tau 0N4R with a 3X Flag tag for Immunoprecipitation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5726)
  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 5726)
  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_Linear Tau 0N4R 3X Flag tag
FEATURES             Location/Qualifiers
     source          1..5726
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..85
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     77..94
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     78..94
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        255..634
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        635..838
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     789..809
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     883..902
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        883..901
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      937..946
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             946..1011
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2202..2218)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2202..2218)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2229..2851
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2428..2447)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2875..2895)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2880..3209
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3060..3195
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3286..3303)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3457..4045)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3537..3556)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4216..5007,5008..5076))
                     /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     4839..4858
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5077..5181)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5208..5663
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5295..5314)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5505..5526
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
       61 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctgatctata cattgaatca
      121 atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata ttggctattg
      181 gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat
      241 atgaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
      301 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      361 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      421 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      481 cttggcagta catcaagtgt atcatatgcc aagtccgccc cctattgacg tcaatgacgg
      541 taaatggccc gcctggcatt atgcccagta catgacctta cgggactttc ctacttggca
      601 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacaccaa
      661 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      721 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta ataaccccgc
      781 cccgttgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct
      841 ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag
      901 ggagacccaa gctggctagc gtttaaactt aagcttgccg ccatggacta caaggaccac
      961 gacggtgatt acaaggatca cgacatcgat tacaaggacg acgacgacaa ggctgagccc
     1021 cgccaggagt tcgaagtgat ggaagatcac gctgggacgt acgggttggg ggacaggaaa
     1081 gatcaggggg gctacaccat gcaccaagac caagagggtg acacggacgc tggcctgaaa
     1141 gctgaagaag caggcattgg agacaccccc agcctggaag acgaagctgc tggtcacgtg
     1201 acccaagctc gcatggtcag taaaagcaaa gacgggactg gaagcgatga caaaaaagcc
     1261 aagggggctg atggtaaaac gaagatcgcc acaccgcggg gagcagcccc tccaggccag
     1321 aagggccagg ccaacgccac caggattcca gcaaaaaccc cgcccgctcc aaagacacca
     1381 cccagctctg gtgaacctcc aaaatcaggg gatcgcagcg gctacagcag ccccggctcc
     1441 ccaggcactc ccggcagccg ctcccgcacc ccgtcccttc caaccccacc cacccgggag
     1501 cccaagaagg tggcagtggt ccgtactcca cccaagtcgc cgtcttccgc caagagccgc
     1561 ctgcagacag cccccgtgcc catgccagac ctgaagaatg tcaagtccaa gatcggctcc
     1621 actgagaacc tgaagcacca gccgggaggc gggaaggtgc agataattaa taagaagctg
     1681 gatcttagca acgtccagtc caagtgtggc tcaaaggata atatcaaaca cgtcccggga
     1741 ggcggcagtg tgcaaatagt ctacaaacca gttgacctga gcaaggtgac ctccaagtgt
     1801 ggctcattag gcaacatcca tcataaacca ggaggtggcc aggtggaagt aaaatctgag
     1861 aagcttgact tcaaggacag agtccagtcg aagattgggt ccctggacaa tatcacccac
     1921 gtccctggcg gaggaaataa aaagattgaa acccacaagc tgaccttccg cgagaacgcc
     1981 aaagccaaga cagaccacgg ggcggagatc gtgtacaagt cgccagtggt gtctggggac
     2041 acgtctccac ggcatctcag caatgtctcc tccaccggca gcatcgacat ggtagactcg
     2101 ccccagctcg ccacgctagc tgacgaggtg tctgcctccc tggccaagca gggtttgtga
     2161 ggtaccgata tcaagcttgt cgactctaga ttgcggccgc ggtcatagct gtttcctgaa
     2221 cagatcccgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     2281 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     2341 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     2401 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     2461 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     2521 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     2581 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     2641 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     2701 tccttctgat tttaaaataa ctataccagc aggaggacgt ccagacacag cataggctac
     2761 ctggccatgc ccaaccggtg ggacatttga gttgcttgct tggcactgtc ctctcatgcg
     2821 ttgggtccac tcagtagatg cctgttgaat tgggtacgcg gccagcttgg ctgtggaatg
     2881 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2941 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3001 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3061 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3121 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3181 gcttttttgg aggcctaggc ttttgcaaaa agctcctcga ctgcattaat gaatcggcca
     3241 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3301 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3361 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3421 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3481 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3541 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3601 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3661 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3721 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3781 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3841 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3901 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3961 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4021 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4081 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4141 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4201 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4261 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4321 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4381 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4441 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4501 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4561 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4621 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4681 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4741 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4801 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4861 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4921 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4981 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5041 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5101 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5161 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag
     5221 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     5281 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5341 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     5401 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     5461 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5521 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     5581 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5641 caaaatatta acgcttacaa tttccattcg ccattcaggc tgcgcaactg ttgggaaggg
     5701 cgatcggtgc gggcctcttc gctatt
//
