LOCUS       sequence-401523-        8051 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For transfection and in vitro assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8051)
  AUTHORS   Margvelani G, Welden JR, Maquera AA, Van Eyk JE, Murray C, Miranda 
            Sardon SC, Stamm S
  TITLE     Influence of FTDP-17 mutants on circular tau RNAs.
  JOURNAL   Biochim Biophys Acta Mol Basis Dis. 2024 Apr;1870(4):167036. doi: 
            10.1016/j.bbadis.2024.167036. Epub 2024 Jan 28.
  PUBMED    38286213
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"; journalName: "Biochim 
            Biophys Acta Mol Basis Dis."; date: "2024-04"; volume: 
            "1870(4):167036. doi"; pages: " 10.1016/j.bbadis.2024.167036. Epub 
            2024 Jan 28"
COMMENT     Sequence Label: Tau BA 12->10 N296N
FEATURES             Location/Qualifiers
     source          1..8051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(82..102)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        87..416
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      267..402
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             483..1277
                     /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(537..556)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1147..1166
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1451..1572
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1488..1507)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1542..1561
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1621..1637)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1621..1637)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1634..1656)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1645..1661
                     /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(1669..1675,1676..1693,1694..1699))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1714..1735
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1852..1869)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2023..2611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2103..2122)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2782..3573,3574..3642))
                     /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     3405..3424
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3643..3747)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3822..3841)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4013..4392
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4393..4596
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4547..4567
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4641..4660
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4641..4659
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6080..6145
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7027..7038
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(7420..7437)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7426..7650
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(7696..8051,1..73)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7783..7802)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7993..8014
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
       61 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
      121 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
      181 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
      241 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
      301 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
      361 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
      421 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
      481 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
      541 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
      601 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
      661 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
      721 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
      781 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
      841 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
      901 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
      961 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     1021 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     1081 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     1141 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     1201 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     1261 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     1321 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     1381 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     1441 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     1501 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     1561 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     1621 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     1681 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     1741 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1801 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1861 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1921 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1981 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2041 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2101 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2161 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2221 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2281 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2341 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2401 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2461 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2521 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2581 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2641 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2701 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2761 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2821 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2881 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     2941 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3001 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3061 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3121 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3181 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3241 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3301 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3361 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3421 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3481 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3541 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3601 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3661 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3721 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     3781 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     3841 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     3901 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     3961 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
     4021 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     4081 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     4141 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     4201 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     4261 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     4321 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     4381 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     4441 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     4501 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     4561 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
     4621 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc tggctagcgt
     4681 ttaaacttaa gcttggtacc gagctcggat ccactagtcc agtgtggtgg aattcccagg
     4741 ctgatggggc ctccagggca gtggtttcca cccagcattc tcaaaagagc cactgaggtc
     4801 tcagtgccat tttcaagatt tcggaagcgg cctgggcacg gctggtcctt cactgggatc
     4861 accacttggc aattatttac acctgagacg aataaaaacc agagtgctga gattacaggc
     4921 atggtggctt acgcttgtaa tcggctttgg gaagccgagg tgggctgatt gcttgagccc
     4981 aggagtttca aactatcctg gacaacatag catgacctcg tctctacaaa aaatacaaaa
     5041 aatttgccag gtgtggtggc atgtgcctgt ggtcccagct acttgggagg ctgaagtagg
     5101 agaatcccct gagccctggg aagtcgaggc tgcactgagc cgtgatgatg tcactgcact
     5161 ccagcctggg tgacaaagtg agaccctatc tcacaaagaa aaaaaacaaa acaaaaaacc
     5221 caaagcacac tgtttccact gtttccagag ttcctgagag gaaaggtcac cgggtgagga
     5281 agacgttctc actgatctgg cagagaaaat gtccagtttt tccaactccc taaaccatgg
     5341 ttttctattt cacagccccc ccttgcaggg acagagccct cgtacagaag ggacacccca
     5401 catttgtctt ccccacaaag cggcctgtgt cctgcctacg gggtcagggc ttctcaaacc
     5461 tggctgtgtg tcagaatcac caggggaact tttcaaaact agagagactg aagccagact
     5521 cctagattct aattctaggt cagggctagg ggctgagatt gtaaaaatcc acaggtgatt
     5581 ctgatgcccg gcaggcttga gaacagccgc agggagttct ctgggaatgt gccggtgggt
     5641 ctagccaggt gtgagtggag atgccgggga acttcctatt actcactcgt cagtgtggcc
     5701 gaacacattt ttcacttgac ctcaggctgg tgaacgctcc cctctggggt tcaggcctca
     5761 cgatgccatc cttttgtgaa gtgaggacct gcaatcccag cttcgtaaag cccgctggaa
     5821 atcactcaca cttctgggat gccttcagag cagccctcta tcccttcagc tcccctggga
     5881 tgtgactcga cctcccgtca ctccccagac tgcctctgcc aagtccgaaa gtggaggcat
     5941 ccttgcgagc aagtaggcgg gtccagggtg gcgcatgtca ctcatcgaaa gtggaggcgt
     6001 ccttgcgagc aagcaggcgg gtccagggtg gcgtgtcact catccttttt tctggctacc
     6061 aaaggtgcag ataattaatg actacaaaga ccatgacggt gattataaag atcatgacat
     6121 cgattacaag gatgacgatg acaagaagaa gctggatctt agcaacgtcc agtccaagtg
     6181 tggctcaaag gataacatca aacacgtccc gggaggcggc agtgtgcaaa tagtctacaa
     6241 accagttgac ctgagcaagg tgacctccaa gtgtggctca ttaggcaaca tccatcataa
     6301 accaggaggt ggccaggtgg aagtaaaatc tgagaagctt gacttcaagg acagagtcca
     6361 gtcgaagatt gggtccctgg acaatatcac ccacgtccct ggcggaggaa ataaaaaggt
     6421 aaagggggta gggtgggttg gatgctgccc ttgggtatat gggcattaat caagttgagt
     6481 ggacaaaggc tggtccagtt cccagaggag gaaaacagag gcttctgtgt tgactggctg
     6541 gatgtgggcc ctcagcagca tccagtgggt ctccactgcc tgtctcaatc acctggagct
     6601 ttagcacgtt tcacacctgg gccccaacct ggagaggctg accaatgggt ctcaggggca
     6661 gctcggttgc tggagttttt gtttttattt atttttatgt atttaaggca gggtctctgt
     6721 attagtccat tctcacactg ctaataaaga catacccaag actgggtaat ttataaagga
     6781 aagaggttta atggactcac agttccatat ggctggggag gcctcaaaat catggcggaa
     6841 ggcaaaggag aagcaaaggc atgtcttaca tggcaacagg caagagagcg tgtgcagggg
     6901 aactcccatt tataaaacca tcagacctca tgagatttat tcactatcat gagaacagca
     6961 tgggaaagac ccgcccccat gattcagtta cctcccactg ggtccctccc atgacacatg
     7021 gaattatggg agctacaatt caagatgaga tttgggtggg gacacagcca aaccatatca
     7081 gtctccctct gtcatccagg ctggagtgca ctggcatgat ctcggctcac tgcagcctct
     7141 acctccctgg gtcaggtgat cttcccacct cagcctccca ggtagctgga actacaggta
     7201 cctgccacta tgcctggcta aatattttgt atttcctgtg gagacgaggt tttgccacgt
     7261 tgcccaggct ggtcttgaac tcctgaggtc aagcaatatg cccacctcgg cctcccaagg
     7321 tgctgggatt acaggtgtga gccacagtgc tcggcctaag tcactgcagt ttttagcggc
     7381 cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     7441 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     7501 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     7561 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     7621 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     7681 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     7741 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     7801 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     7861 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     7921 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     7981 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     8041 attcttttga t
//
