LOCUS       sequence-381821-        7398 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insert contains intronic ZKSCAN1 Alu repeat elements flanked by the 
            wild-type tau cDNA exons 7,9-12. Expresses the tau circRNA 12-->7 WT
            with 3X flag tag in exon 7..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7398)
  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 7398)
  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_ZKSCAN1_Tau7-12WT
FEATURES             Location/Qualifiers
     source          1..7398
                     /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             1417..1482
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2989..3006)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2995..3219
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3265..3693
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3352..3371)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3562..3583
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3702..3722)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3707..4036
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3887..4022
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4103..4897
                     /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(4157..4176)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4767..4786
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5071..5192
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5108..5127)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5162..5181
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5241..5257)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5241..5257)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5254..5276)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5265..5281
                     /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(5289..5295,5296..5313,5314..5319))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5334..5355
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5472..5489)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5643..6231)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5723..5742)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6402..7193,7194..7262))
                     /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     7025..7044
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7263..7367)
                     /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 ggaattcagt
      961 gacagtggag attgtacagt tttttcctcg atttgtcagg attttttttt tttgacggag
     1021 tttaacttct tgtctcccag gtaggaagtg cagtggcgta atctcggctc actacaacct
     1081 ccacctcctg ggttcaagcg tttctcctgc ctcagctttc cgagtagctg ggattacagg
     1141 cgcctgccac catgccctgc tgacttttgt atttttagta gagacggggt ttcaccatgt
     1201 tggccaggct ggtcttgaac tcctgaccgc aggcgattgg cctgcctcgg cctcccaaag
     1261 tgctgagatt acaggcgtga gccaccaccc ccggcctcag gagcgttctg atagtgcctc
     1321 gatgtgctgc ctcctataaa gtgttagcag cacagatcac tttttgtaaa ggtacgtact
     1381 aatgactttt tttttatact tcagggggct gatggtgact acaaagacca tgacggtgat
     1441 tataaagatc atgacatcga ttacaaggat gacgatgaca agaaaacgaa gatcgccaca
     1501 ccgcggggag cagcccctcc aggccagaag ggccaggcca acgccaccag gattccagca
     1561 aaaaccccgc ccgctccaaa gacaccaccc agctctggtg aacctccaaa atcaggggat
     1621 cgcagcggct acagcagccc cggctcccca ggcactcccg gcagccgctc ccgcaccccg
     1681 tcccttccaa ccccacccac ccgggagccc aagaaggtgg cagtggtccg tactccaccc
     1741 aagtcgccgt cttccgccaa gagccgcctg cagacagccc ccgtgcccat gccagacctg
     1801 aagaatgtca agtccaagat cggctccact gagaacctga agcaccagcc gggaggcggg
     1861 aaggtgcaga taattaataa gaagctggat cttagcaacg tccagtccaa gtgtggctca
     1921 aaggataata tcaaacacgt cccgggaggc ggcagtgtgc aaatagtcta caaaccagtt
     1981 gacctgagca aggtgacctc caagtgtggc tcattaggca acatccatca taaaccagga
     2041 ggtggccagg tggaagtaaa atctgagaag cttgacttca aggacagagt ccagtcgaag
     2101 attgggtccc tggacaatat cacccacgtc cctggcggag gaaataaaaa ggtaagaagc
     2161 aaggtttcat ttaggggaag ggaaatgatt caggacgaga gtctttgtgc tgctgagtgc
     2221 ctgtgatgaa gaagcatgtt agtcctgggc aacgtagcga gaccccatct ctacaaaaaa
     2281 tagaaaaatt agccaggtat agtggcgcac acctgtgatt ccagctacgc aggaggctga
     2341 ggtgggagga ttgcttgagc ccaggaggtt gaggctgcag tgagctggaa tcatgccact
     2401 actccaacct gggcaacaca gcaaggaccc tgtctcaaaa gctacttaca gaaaagaatt
     2461 aggctcggca cggtagctca cacctgtaat cccagcactt tgggaggctg aggcgggcag
     2521 atcacttgag gtcaggagtt tgagaccagc ctggccaaca tggtgaaacc ttgtctctac
     2581 taaaaatatg aaaattagcc aggcatggtg gcacattcct gtaatcccag ctactcggga
     2641 ggctgaggca ggagaatcac ttgaacccag gaggtggagg ttgcagtaag ccgagatcgt
     2701 accactgtgc tctagccttg gtgacagagc gagactgtct taaaaaaaaa aaaaaaaaaa
     2761 gaattaatta aaaatttaaa aaaaaatgaa aaaaagctgc atgcttgttt tttgttttta
     2821 gttattctac attgttgtca ttattaccaa atattgggga aaatacaact tacagaccaa
     2881 tctcaggagt taaatgttac tacgaaggca aatgaactat gcgtaatgaa cctggtaggc
     2941 attagcggcc gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc
     3001 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     3061 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     3121 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     3181 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     3241 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3301 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3361 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3421 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3481 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3541 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3601 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3661 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3721 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3781 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3841 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3901 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3961 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4021 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     4081 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4141 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4201 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4261 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4321 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4381 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4441 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4501 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4561 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4621 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4681 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4741 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4801 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4861 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4921 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4981 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5041 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5101 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5161 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5221 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5281 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5341 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5401 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5461 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5521 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5581 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5641 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5701 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5761 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5821 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5881 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5941 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6001 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6061 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6121 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6181 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6241 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6301 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6361 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6421 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6481 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6541 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6601 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6661 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6721 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6781 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6841 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6901 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6961 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7021 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7081 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7141 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7201 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7261 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7321 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7381 aaagtgccac ctgacgtc
//
