LOCUS       sequence-401581-        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 G303V
FEATURES             Location/Qualifiers
     source          1..8051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(81..101)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        86..415
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      266..401
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             482..1276
                     /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(536..555)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1146..1165
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1450..1571
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1487..1506)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1541..1560
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1620..1636)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1620..1636)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1633..1655)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1644..1660
                     /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(1668..1674,1675..1692,1693..1698))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1713..1734
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1851..1868)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2022..2610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2102..2121)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2781..3572,3573..3641))
                     /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     3404..3423
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3642..3746)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3821..3840)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4012..4391
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4392..4595
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4546..4566
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4640..4659
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4640..4658
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6079..6144
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7026..7037
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(7419..7436)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7425..7649
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(7695..8051,1..72)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7782..7801)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7992..8013
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
       61 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
      121 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
      181 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
      241 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
      301 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
      361 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
      421 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
      481 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
      541 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
      601 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
      661 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
      721 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
      781 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
      841 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
      901 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
      961 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     1021 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     1081 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     1141 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     1201 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     1261 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     1321 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     1381 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     1441 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     1501 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     1561 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     1621 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1681 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1741 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     1801 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1861 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1921 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1981 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2041 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2101 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2161 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2221 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2281 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2341 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2401 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2461 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2521 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2581 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2641 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2701 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2761 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2821 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2881 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     2941 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3001 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3061 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3121 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3181 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3241 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3301 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3361 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3421 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3481 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3541 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3601 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3661 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3721 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     3781 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     3841 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     3901 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     3961 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
     4021 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     4081 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     4141 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     4201 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     4261 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     4321 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     4381 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     4441 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     4501 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     4561 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
     4621 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct ggctagcgtt
     4681 taaacttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga attcccaggc
     4741 tgatggggcc tccagggcag tggtttccac ccagcattct caaaagagcc actgaggtct
     4801 cagtgccatt ttcaagattt cggaagcggc ctgggcacgg ctggtccttc actgggatca
     4861 ccacttggca attatttaca cctgagacga ataaaaacca gagtgctgag attacaggca
     4921 tggtggctta cgcttgtaat cggctttggg aagccgaggt gggctgattg cttgagccca
     4981 ggagtttcaa actatcctgg acaacatagc atgacctcgt ctctacaaaa aatacaaaaa
     5041 atttgccagg tgtggtggca tgtgcctgtg gtcccagcta cttgggaggc tgaagtagga
     5101 gaatcccctg agccctggga agtcgaggct gcactgagcc gtgatgatgt cactgcactc
     5161 cagcctgggt gacaaagtga gaccctatct cacaaagaaa aaaaacaaaa caaaaaaccc
     5221 aaagcacact gtttccactg tttccagagt tcctgagagg aaaggtcacc gggtgaggaa
     5281 gacgttctca ctgatctggc agagaaaatg tccagttttt ccaactccct aaaccatggt
     5341 tttctatttc acagcccccc cttgcaggga cagagccctc gtacagaagg gacaccccac
     5401 atttgtcttc cccacaaagc ggcctgtgtc ctgcctacgg ggtcagggct tctcaaacct
     5461 ggctgtgtgt cagaatcacc aggggaactt ttcaaaacta gagagactga agccagactc
     5521 ctagattcta attctaggtc agggctaggg gctgagattg taaaaatcca caggtgattc
     5581 tgatgcccgg caggcttgag aacagccgca gggagttctc tgggaatgtg ccggtgggtc
     5641 tagccaggtg tgagtggaga tgccggggaa cttcctatta ctcactcgtc agtgtggccg
     5701 aacacatttt tcacttgacc tcaggctggt gaacgctccc ctctggggtt caggcctcac
     5761 gatgccatcc ttttgtgaag tgaggacctg caatcccagc ttcgtaaagc ccgctggaaa
     5821 tcactcacac ttctgggatg ccttcagagc agccctctat cccttcagct cccctgggat
     5881 gtgactcgac ctcccgtcac tccccagact gcctctgcca agtccgaaag tggaggcatc
     5941 cttgcgagca agtaggcggg tccagggtgg cgcatgtcac tcatcgaaag tggaggcgtc
     6001 cttgcgagca agcaggcggg tccagggtgg cgtgtcactc atcctttttt ctggctacca
     6061 aaggtgcaga taattaatga ctacaaagac catgacggtg attataaaga tcatgacatc
     6121 gattacaagg atgacgatga caagaagaag ctggatctta gcaacgtcca gtccaagtgt
     6181 ggctcaaagg ataatatcaa acacgtcccg ggagtcggca gtgtgcaaat agtctacaaa
     6241 ccagttgacc tgagcaaggt gacctccaag tgtggctcat taggcaacat ccatcataaa
     6301 ccaggaggtg gccaggtgga agtaaaatct gagaagcttg acttcaagga cagagtccag
     6361 tcgaagattg ggtccctgga caatatcacc cacgtccctg gcggaggaaa taaaaaggta
     6421 aagggggtag ggtgggttgg atgctgccct tgggtatatg ggcattaatc aagttgagtg
     6481 gacaaaggct ggtccagttc ccagaggagg aaaacagagg cttctgtgtt gactggctgg
     6541 atgtgggccc tcagcagcat ccagtgggtc tccactgcct gtctcaatca cctggagctt
     6601 tagcacgttt cacacctggg ccccaacctg gagaggctga ccaatgggtc tcaggggcag
     6661 ctcggttgct ggagtttttg tttttattta tttttatgta tttaaggcag ggtctctgta
     6721 ttagtccatt ctcacactgc taataaagac atacccaaga ctgggtaatt tataaaggaa
     6781 agaggtttaa tggactcaca gttccatatg gctggggagg cctcaaaatc atggcggaag
     6841 gcaaaggaga agcaaaggca tgtcttacat ggcaacaggc aagagagcgt gtgcagggga
     6901 actcccattt ataaaaccat cagacctcat gagatttatt cactatcatg agaacagcat
     6961 gggaaagacc cgcccccatg attcagttac ctcccactgg gtccctccca tgacacatgg
     7021 aattatggga gctacaattc aagatgagat ttgggtgggg acacagccaa accatatcag
     7081 tctccctctg tcatccaggc tggagtgcac tggcatgatc tcggctcact gcagcctcta
     7141 cctccctggg tcaggtgatc ttcccacctc agcctcccag gtagctggaa ctacaggtac
     7201 ctgccactat gcctggctaa atattttgta tttcctgtgg agacgaggtt ttgccacgtt
     7261 gcccaggctg gtcttgaact cctgaggtca agcaatatgc ccacctcggc ctcccaaggt
     7321 gctgggatta caggtgtgag ccacagtgct cggcctaagt cactgcagtt tttagcggcc
     7381 gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     7441 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     7501 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     7561 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     7621 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     7681 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     7741 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     7801 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     7861 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     7921 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     7981 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     8041 ttcttttgat t
//
