LOCUS       sequence-401582-        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 P301L
FEATURES             Location/Qualifiers
     source          1..8051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(84..104)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        89..418
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      269..404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             485..1279
                     /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(539..558)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1149..1168
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1453..1574
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1490..1509)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1544..1563
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1623..1639)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1623..1639)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1636..1658)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1647..1663
                     /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(1671..1677,1678..1695,1696..1701))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1716..1737
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1854..1871)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2025..2613)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2105..2124)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2784..3575,3576..3644))
                     /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     3407..3426
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3645..3749)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3824..3843)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4015..4394
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4395..4598
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4549..4569
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4643..4662
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4643..4661
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6082..6147
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7029..7040
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(7422..7439)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7428..7652
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(7698..8051,1..75)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7785..7804)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7995..8016
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
       61 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
      121 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
      181 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      241 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
      301 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
      361 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
      421 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
      481 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
      541 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
      601 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
      661 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
      721 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
      781 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
      841 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
      901 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
      961 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     1021 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     1081 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     1141 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     1201 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     1261 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     1321 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     1381 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     1441 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     1501 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     1561 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     1621 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1681 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1741 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1801 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1861 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1921 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1981 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2041 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2101 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2161 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2221 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2281 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2341 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2401 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2461 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2521 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2581 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2641 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2701 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2761 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2821 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2881 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2941 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3001 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3061 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3121 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3181 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3241 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3301 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3361 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3421 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3481 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3541 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3601 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3661 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3721 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     3781 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     3841 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     3901 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     3961 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
     4021 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     4081 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     4141 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     4201 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     4261 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     4321 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     4381 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     4441 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     4501 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     4561 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
     4621 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
     4681 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattccca
     4741 ggctgatggg gcctccaggg cagtggtttc cacccagcat tctcaaaaga gccactgagg
     4801 tctcagtgcc attttcaaga tttcggaagc ggcctgggca cggctggtcc ttcactggga
     4861 tcaccacttg gcaattattt acacctgaga cgaataaaaa ccagagtgct gagattacag
     4921 gcatggtggc ttacgcttgt aatcggcttt gggaagccga ggtgggctga ttgcttgagc
     4981 ccaggagttt caaactatcc tggacaacat agcatgacct cgtctctaca aaaaatacaa
     5041 aaaatttgcc aggtgtggtg gcatgtgcct gtggtcccag ctacttggga ggctgaagta
     5101 ggagaatccc ctgagccctg ggaagtcgag gctgcactga gccgtgatga tgtcactgca
     5161 ctccagcctg ggtgacaaag tgagacccta tctcacaaag aaaaaaaaca aaacaaaaaa
     5221 cccaaagcac actgtttcca ctgtttccag agttcctgag aggaaaggtc accgggtgag
     5281 gaagacgttc tcactgatct ggcagagaaa atgtccagtt tttccaactc cctaaaccat
     5341 ggttttctat ttcacagccc ccccttgcag ggacagagcc ctcgtacaga agggacaccc
     5401 cacatttgtc ttccccacaa agcggcctgt gtcctgccta cggggtcagg gcttctcaaa
     5461 cctggctgtg tgtcagaatc accaggggaa cttttcaaaa ctagagagac tgaagccaga
     5521 ctcctagatt ctaattctag gtcagggcta ggggctgaga ttgtaaaaat ccacaggtga
     5581 ttctgatgcc cggcaggctt gagaacagcc gcagggagtt ctctgggaat gtgccggtgg
     5641 gtctagccag gtgtgagtgg agatgccggg gaacttccta ttactcactc gtcagtgtgg
     5701 ccgaacacat ttttcacttg acctcaggct ggtgaacgct cccctctggg gttcaggcct
     5761 cacgatgcca tccttttgtg aagtgaggac ctgcaatccc agcttcgtaa agcccgctgg
     5821 aaatcactca cacttctggg atgccttcag agcagccctc tatcccttca gctcccctgg
     5881 gatgtgactc gacctcccgt cactccccag actgcctctg ccaagtccga aagtggaggc
     5941 atccttgcga gcaagtaggc gggtccaggg tggcgcatgt cactcatcga aagtggaggc
     6001 gtccttgcga gcaagcaggc gggtccaggg tggcgtgtca ctcatccttt tttctggcta
     6061 ccaaaggtgc agataattaa tgactacaaa gaccatgacg gtgattataa agatcatgac
     6121 atcgattaca aggatgacga tgacaagaag aagctggatc ttagcaacgt ccagtccaag
     6181 tgtggctcaa aggataatat caaacacgtc ctgggaggcg gcagtgtgca aatagtctac
     6241 aaaccagttg acctgagcaa ggtgacctcc aagtgtggct cattaggcaa catccatcat
     6301 aaaccaggag gtggccaggt ggaagtaaaa tctgagaagc ttgacttcaa ggacagagtc
     6361 cagtcgaaga ttgggtccct ggacaatatc acccacgtcc ctggcggagg aaataaaaag
     6421 gtaaaggggg tagggtgggt tggatgctgc ccttgggtat atgggcatta atcaagttga
     6481 gtggacaaag gctggtccag ttcccagagg aggaaaacag aggcttctgt gttgactggc
     6541 tggatgtggg ccctcagcag catccagtgg gtctccactg cctgtctcaa tcacctggag
     6601 ctttagcacg tttcacacct gggccccaac ctggagaggc tgaccaatgg gtctcagggg
     6661 cagctcggtt gctggagttt ttgtttttat ttatttttat gtatttaagg cagggtctct
     6721 gtattagtcc attctcacac tgctaataaa gacataccca agactgggta atttataaag
     6781 gaaagaggtt taatggactc acagttccat atggctgggg aggcctcaaa atcatggcgg
     6841 aaggcaaagg agaagcaaag gcatgtctta catggcaaca ggcaagagag cgtgtgcagg
     6901 ggaactccca tttataaaac catcagacct catgagattt attcactatc atgagaacag
     6961 catgggaaag acccgccccc atgattcagt tacctcccac tgggtccctc ccatgacaca
     7021 tggaattatg ggagctacaa ttcaagatga gatttgggtg gggacacagc caaaccatat
     7081 cagtctccct ctgtcatcca ggctggagtg cactggcatg atctcggctc actgcagcct
     7141 ctacctccct gggtcaggtg atcttcccac ctcagcctcc caggtagctg gaactacagg
     7201 tacctgccac tatgcctggc taaatatttt gtatttcctg tggagacgag gttttgccac
     7261 gttgcccagg ctggtcttga actcctgagg tcaagcaata tgcccacctc ggcctcccaa
     7321 ggtgctggga ttacaggtgt gagccacagt gctcggccta agtcactgca gtttttagcg
     7381 gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     7441 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     7501 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     7561 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     7621 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     7681 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     7741 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     7801 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     7861 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     7921 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     7981 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     8041 ctattctttt g
//
