LOCUS       sequence-401584-        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 V393I
FEATURES             Location/Qualifiers
     source          1..8051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             111..905
                     /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(165..184)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     775..794
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1079..1200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1116..1135)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1170..1189
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1249..1265)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1249..1265)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1262..1284)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1273..1289
                     /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(1297..1303,1304..1321,1322..1327))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1342..1363
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1480..1497)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1651..2239)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1731..1750)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2410..3201,3202..3270))
                     /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     3033..3052
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3271..3375)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3450..3469)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3641..4020
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4021..4224
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4175..4195
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4269..4288
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4269..4287
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5708..5773
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6655..6666
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(7048..7065)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7054..7278
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7324..7752
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7411..7430)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7621..7642
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7761..7781)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(7766..8051,1..44)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7946..8051,1..30)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
       61 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
      121 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
      181 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
      241 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
      301 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
      361 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
      421 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
      481 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
      541 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
      601 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
      661 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
      721 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
      781 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
      841 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
      901 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
      961 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     1021 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     1081 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     1141 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     1201 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     1261 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     1321 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     1381 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     1441 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     1501 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     1561 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     1621 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1681 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1741 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1801 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1861 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1921 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1981 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2041 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2101 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2161 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2221 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2281 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2341 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2401 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2461 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2521 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     2581 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2641 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2701 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     2761 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     2821 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2881 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     2941 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3001 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3061 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     3121 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3181 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3241 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3301 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3361 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     3421 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3481 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     3541 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
     3601 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
     3661 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     3721 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     3781 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     3841 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     3901 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     3961 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     4021 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     4081 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     4141 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     4201 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
     4261 atcgaaatta atacgactca ctatagggag acccaagctg gctagcgttt aaacttaagc
     4321 ttggtaccga gctcggatcc actagtccag tgtggtggaa ttcccaggct gatggggcct
     4381 ccagggcagt ggtttccacc cagcattctc aaaagagcca ctgaggtctc agtgccattt
     4441 tcaagatttc ggaagcggcc tgggcacggc tggtccttca ctgggatcac cacttggcaa
     4501 ttatttacac ctgagacgaa taaaaaccag agtgctgaga ttacaggcat ggtggcttac
     4561 gcttgtaatc ggctttggga agccgaggtg ggctgattgc ttgagcccag gagtttcaaa
     4621 ctatcctgga caacatagca tgacctcgtc tctacaaaaa atacaaaaaa tttgccaggt
     4681 gtggtggcat gtgcctgtgg tcccagctac ttgggaggct gaagtaggag aatcccctga
     4741 gccctgggaa gtcgaggctg cactgagccg tgatgatgtc actgcactcc agcctgggtg
     4801 acaaagtgag accctatctc acaaagaaaa aaaacaaaac aaaaaaccca aagcacactg
     4861 tttccactgt ttccagagtt cctgagagga aaggtcaccg ggtgaggaag acgttctcac
     4921 tgatctggca gagaaaatgt ccagtttttc caactcccta aaccatggtt ttctatttca
     4981 cagccccccc ttgcagggac agagccctcg tacagaaggg acaccccaca tttgtcttcc
     5041 ccacaaagcg gcctgtgtcc tgcctacggg gtcagggctt ctcaaacctg gctgtgtgtc
     5101 agaatcacca ggggaacttt tcaaaactag agagactgaa gccagactcc tagattctaa
     5161 ttctaggtca gggctagggg ctgagattgt aaaaatccac aggtgattct gatgcccggc
     5221 aggcttgaga acagccgcag ggagttctct gggaatgtgc cggtgggtct agccaggtgt
     5281 gagtggagat gccggggaac ttcctattac tcactcgtca gtgtggccga acacattttt
     5341 cacttgacct caggctggtg aacgctcccc tctggggttc aggcctcacg atgccatcct
     5401 tttgtgaagt gaggacctgc aatcccagct tcgtaaagcc cgctggaaat cactcacact
     5461 tctgggatgc cttcagagca gccctctatc ccttcagctc ccctgggatg tgactcgacc
     5521 tcccgtcact ccccagactg cctctgccaa gtccgaaagt ggaggcatcc ttgcgagcaa
     5581 gtaggcgggt ccagggtggc gcatgtcact catcgaaagt ggaggcgtcc ttgcgagcaa
     5641 gcaggcgggt ccagggtggc gtgtcactca tccttttttc tggctaccaa aggtgcagat
     5701 aattaatgac tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga
     5761 tgacgatgac aagaagaagc tggatcttag caacgtccag tccaagtgtg gctcaaagga
     5821 taatatcaaa cacgtcccgg gaggcggcag tgtgcaaata gtctacaaac cagttgacct
     5881 gagcaaggtg acctccaagt gtggctcatt aggcaacatc catcataaac caggaggtgg
     5941 ccaggtggaa gtaaaatctg agaagcttga cttcaaggac agagtccagt cgaagattgg
     6001 gtccctggac aatatcaccc acatccctgg cggaggaaat aaaaaggtaa agggggtagg
     6061 gtgggttgga tgctgccctt gggtatatgg gcattaatca agttgagtgg acaaaggctg
     6121 gtccagttcc cagaggagga aaacagaggc ttctgtgttg actggctgga tgtgggccct
     6181 cagcagcatc cagtgggtct ccactgcctg tctcaatcac ctggagcttt agcacgtttc
     6241 acacctgggc cccaacctgg agaggctgac caatgggtct caggggcagc tcggttgctg
     6301 gagtttttgt ttttatttat ttttatgtat ttaaggcagg gtctctgtat tagtccattc
     6361 tcacactgct aataaagaca tacccaagac tgggtaattt ataaaggaaa gaggtttaat
     6421 ggactcacag ttccatatgg ctggggaggc ctcaaaatca tggcggaagg caaaggagaa
     6481 gcaaaggcat gtcttacatg gcaacaggca agagagcgtg tgcaggggaa ctcccattta
     6541 taaaaccatc agacctcatg agatttattc actatcatga gaacagcatg ggaaagaccc
     6601 gcccccatga ttcagttacc tcccactggg tccctcccat gacacatgga attatgggag
     6661 ctacaattca agatgagatt tgggtgggga cacagccaaa ccatatcagt ctccctctgt
     6721 catccaggct ggagtgcact ggcatgatct cggctcactg cagcctctac ctccctgggt
     6781 caggtgatct tcccacctca gcctcccagg tagctggaac tacaggtacc tgccactatg
     6841 cctggctaaa tattttgtat ttcctgtgga gacgaggttt tgccacgttg cccaggctgg
     6901 tcttgaactc ctgaggtcaa gcaatatgcc cacctcggcc tcccaaggtg ctgggattac
     6961 aggtgtgagc cacagtgctc ggcctaagtc actgcagttt ttagcggccg ctcgagtcta
     7021 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     7081 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     7141 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     7201 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     7261 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     7321 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     7381 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     7441 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     7501 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     7561 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     7621 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     7681 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     7741 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     7801 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     7861 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     7921 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     7981 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     8041 tgagctattc c
//
