LOCUS       sequence-401575-        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 P301S
FEATURES             Location/Qualifiers
     source          1..8051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(27..356)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      41..176
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     341..361
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(370..798)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(480..501)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     692..711
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    844..1068
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1057..1074
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1456..1467)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             complement(2349..2414)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3834..3853)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3835..3853)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3898..4101)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3927..3947)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4102..4481
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4653..4672
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4747..4851
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4852..4920,4921..5712)
                     /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     complement(5070..5089)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5883..6471
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6372..6391
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6625..6642
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6759..6780
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6795..6800,6801..6818,6819..6825)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6833..6849
                     /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)."
     primer_bind     6838..6860
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6857..6873
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6857..6873
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6922..7043
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6933..6952)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6987..7006
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(7217..8011)
                     /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(7328..7347)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7938..7957
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 atggatatac aagctcccgg gagctttttg caaaagccta ggcctccaaa aaagcctcct
       61 cactacttct ggaatagctc agaggcagag gcggcctcgg cctctgcata aataaaaaaa
      121 attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc
      181 ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg
      241 cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt
      301 tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc
      361 cacagaatta attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
      421 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
      481 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
      541 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
      601 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
      661 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
      721 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
      781 cgccgctaca gggcgcgtgg ggataccccc tagagcccca gctggttctt tccgcctcag
      841 aagccataga gcccaccgca tccccagcat gcctgctatt gtcttcccaa tcctccccct
      901 tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga caatgcgatg
      961 caatttcctc attttattag gaaaggacag tgggagtggc accttccagg gtcaaggaag
     1021 gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagtc gaggctgatc
     1081 agcgggttta aacgggccct ctagactcga gcggccgcta aaaactgcag tgacttaggc
     1141 cgagcactgt ggctcacacc tgtaatccca gcaccttggg aggccgaggt gggcatattg
     1201 cttgacctca ggagttcaag accagcctgg gcaacgtggc aaaacctcgt ctccacagga
     1261 aatacaaaat atttagccag gcatagtggc aggtacctgt agttccagct acctgggagg
     1321 ctgaggtggg aagatcacct gacccaggga ggtagaggct gcagtgagcc gagatcatgc
     1381 cagtgcactc cagcctggat gacagaggga gactgatatg gtttggctgt gtccccaccc
     1441 aaatctcatc ttgaattgta gctcccataa ttccatgtgt catgggaggg acccagtggg
     1501 aggtaactga atcatggggg cgggtctttc ccatgctgtt ctcatgatag tgaataaatc
     1561 tcatgaggtc tgatggtttt ataaatggga gttcccctgc acacgctctc ttgcctgttg
     1621 ccatgtaaga catgcctttg cttctccttt gccttccgcc atgattttga ggcctcccca
     1681 gccatatgga actgtgagtc cattaaacct ctttccttta taaattaccc agtcttgggt
     1741 atgtctttat tagcagtgtg agaatggact aatacagaga ccctgcctta aatacataaa
     1801 aataaataaa aacaaaaact ccagcaaccg agctgcccct gagacccatt ggtcagcctc
     1861 tccaggttgg ggcccaggtg tgaaacgtgc taaagctcca ggtgattgag acaggcagtg
     1921 gagacccact ggatgctgct gagggcccac atccagccag tcaacacaga agcctctgtt
     1981 ttcctcctct gggaactgga ccagcctttg tccactcaac ttgattaatg cccatatacc
     2041 caagggcagc atccaaccca ccctaccccc tttacctttt tatttcctcc gccagggacg
     2101 tgggtgatat tgtccaggga cccaatcttc gactggactc tgtccttgaa gtcaagcttc
     2161 tcagatttta cttccacctg gccacctcct ggtttatgat ggatgttgcc taatgagcca
     2221 cacttggagg tcaccttgct caggtcaact ggtttgtaga ctatttgcac actgccgcct
     2281 cccgagacgt gtttgatatt atcctttgag ccacacttgg actggacgtt gctaagatcc
     2341 agcttcttct tgtcatcgtc atccttgtaa tcgatgtcat gatctttata atcaccgtca
     2401 tggtctttgt agtcattaat tatctgcacc tttggtagcc agaaaaaagg atgagtgaca
     2461 cgccaccctg gacccgcctg cttgctcgca aggacgcctc cactttcgat gagtgacatg
     2521 cgccaccctg gacccgccta cttgctcgca aggatgcctc cactttcgga cttggcagag
     2581 gcagtctggg gagtgacggg aggtcgagtc acatcccagg ggagctgaag ggatagaggg
     2641 ctgctctgaa ggcatcccag aagtgtgagt gatttccagc gggctttacg aagctgggat
     2701 tgcaggtcct cacttcacaa aaggatggca tcgtgaggcc tgaaccccag aggggagcgt
     2761 tcaccagcct gaggtcaagt gaaaaatgtg ttcggccaca ctgacgagtg agtaatagga
     2821 agttccccgg catctccact cacacctggc tagacccacc ggcacattcc cagagaactc
     2881 cctgcggctg ttctcaagcc tgccgggcat cagaatcacc tgtggatttt tacaatctca
     2941 gcccctagcc ctgacctaga attagaatct aggagtctgg cttcagtctc tctagttttg
     3001 aaaagttccc ctggtgattc tgacacacag ccaggtttga gaagccctga ccccgtaggc
     3061 aggacacagg ccgctttgtg gggaagacaa atgtggggtg tcccttctgt acgagggctc
     3121 tgtccctgca agggggggct gtgaaataga aaaccatggt ttagggagtt ggaaaaactg
     3181 gacattttct ctgccagatc agtgagaacg tcttcctcac ccggtgacct ttcctctcag
     3241 gaactctgga aacagtggaa acagtgtgct ttgggttttt tgttttgttt tttttctttg
     3301 tgagataggg tctcactttg tcacccaggc tggagtgcag tgacatcatc acggctcagt
     3361 gcagcctcga cttcccaggg ctcaggggat tctcctactt cagcctccca agtagctggg
     3421 accacaggca catgccacca cacctggcaa attttttgta ttttttgtag agacgaggtc
     3481 atgctatgtt gtccaggata gtttgaaact cctgggctca agcaatcagc ccacctcggc
     3541 ttcccaaagc cgattacaag cgtaagccac catgcctgta atctcagcac tctggttttt
     3601 attcgtctca ggtgtaaata attgccaagt ggtgatccca gtgaaggacc agccgtgccc
     3661 aggccgcttc cgaaatcttg aaaatggcac tgagacctca gtggctcttt tgagaatgct
     3721 gggtggaaac cactgccctg gaggccccat cagcctggga attccaccac actggactag
     3781 tggatccgag ctcggtacca agcttaagtt taaacgctag ccagcttggg tctccctata
     3841 gtgagtcgta ttaatttcga taagccagta agcagtgggt tctctagtta gccagagagc
     3901 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga
     3961 gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     4021 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     4081 gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca
     4141 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     4201 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     4261 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     4321 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     4381 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     4441 cgtaattgat tactattaat aactagtcaa taatcaatgt caacgcgtat atctggcccg
     4501 tacatcgcga agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc aattgtcggt
     4561 caagccttgc cttgttgtag cttaaatttt gctcgcgcac tactcagcga cctccaacac
     4621 acaagcaggg agcagatact ggcttaacta tgcggcatca gagcagattg tactgagagt
     4681 gcaccatagg ggatcgggag atctcccgat ccgtcgacgt caggtggcac ttttcgggga
     4741 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4801 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4861 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4921 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4981 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5041 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5101 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5161 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5221 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5281 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5341 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5401 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5461 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5521 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5581 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5641 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5701 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5761 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5821 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5881 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5941 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6001 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6061 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6121 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6181 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6241 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6301 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6361 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6421 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6481 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     6541 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     6601 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     6661 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     6721 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     6781 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     6841 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag
     6901 gtcgacggta tacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     6961 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     7021 ttataagctg caataaacaa gttggggtgg gcgaagaact ccagcatgag atccccgcgc
     7081 tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag
     7141 gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg
     7201 aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     7261 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     7321 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     7381 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     7441 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg agcctggcga
     7501 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     7561 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     7621 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     7681 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     7741 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     7801 ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg
     7861 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     7921 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     7981 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     8041 cagatccgaa a
//
