LOCUS       sequence-401574-        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 K369I
FEATURES             Location/Qualifiers
     source          1..8051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(108..128)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        113..442
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      293..428
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             509..1303
                     /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(563..582)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1173..1192
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1477..1598
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1514..1533)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1568..1587
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1647..1663)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1647..1663)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1660..1682)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1671..1687
                     /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(1695..1701,1702..1719,1720..1725))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1740..1761
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1878..1895)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2049..2637)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2129..2148)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2808..3599,3600..3668))
                     /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     3431..3450
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3669..3773)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3848..3867)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4039..4418
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4419..4622
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4573..4593
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4667..4686
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4667..4685
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6106..6171
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7053..7064
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(7446..7463)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7452..7676
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(7722..8051,1..99)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7809..7828)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8019..8040
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
       61 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
      121 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
      181 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      241 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
      301 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
      361 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
      421 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
      481 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
      541 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
      601 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
      661 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
      721 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
      781 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
      841 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
      901 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
      961 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     1021 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     1081 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     1141 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     1201 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     1261 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     1321 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     1381 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     1441 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     1501 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1561 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     1621 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1681 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1741 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1801 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1861 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1921 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1981 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2041 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2101 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2161 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2221 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2281 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2341 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2401 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2461 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2521 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2581 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2641 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2701 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2761 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2821 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2881 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2941 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3001 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3061 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3121 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3181 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3241 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3301 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3361 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3421 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3481 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3541 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3601 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3661 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3721 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3781 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     3841 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     3901 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     3961 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     4021 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     4081 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     4141 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     4201 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     4261 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     4321 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     4381 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     4441 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     4501 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     4561 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     4621 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
     4681 atagggagac ccaagctggc tagcgtttaa acttaagctt ggtaccgagc tcggatccac
     4741 tagtccagtg tggtggaatt cccaggctga tggggcctcc agggcagtgg tttccaccca
     4801 gcattctcaa aagagccact gaggtctcag tgccattttc aagatttcgg aagcggcctg
     4861 ggcacggctg gtccttcact gggatcacca cttggcaatt atttacacct gagacgaata
     4921 aaaaccagag tgctgagatt acaggcatgg tggcttacgc ttgtaatcgg ctttgggaag
     4981 ccgaggtggg ctgattgctt gagcccagga gtttcaaact atcctggaca acatagcatg
     5041 acctcgtctc tacaaaaaat acaaaaaatt tgccaggtgt ggtggcatgt gcctgtggtc
     5101 ccagctactt gggaggctga agtaggagaa tcccctgagc cctgggaagt cgaggctgca
     5161 ctgagccgtg atgatgtcac tgcactccag cctgggtgac aaagtgagac cctatctcac
     5221 aaagaaaaaa aacaaaacaa aaaacccaaa gcacactgtt tccactgttt ccagagttcc
     5281 tgagaggaaa ggtcaccggg tgaggaagac gttctcactg atctggcaga gaaaatgtcc
     5341 agtttttcca actccctaaa ccatggtttt ctatttcaca gccccccctt gcagggacag
     5401 agccctcgta cagaagggac accccacatt tgtcttcccc acaaagcggc ctgtgtcctg
     5461 cctacggggt cagggcttct caaacctggc tgtgtgtcag aatcaccagg ggaacttttc
     5521 aaaactagag agactgaagc cagactccta gattctaatt ctaggtcagg gctaggggct
     5581 gagattgtaa aaatccacag gtgattctga tgcccggcag gcttgagaac agccgcaggg
     5641 agttctctgg gaatgtgccg gtgggtctag ccaggtgtga gtggagatgc cggggaactt
     5701 cctattactc actcgtcagt gtggccgaac acatttttca cttgacctca ggctggtgaa
     5761 cgctcccctc tggggttcag gcctcacgat gccatccttt tgtgaagtga ggacctgcaa
     5821 tcccagcttc gtaaagcccg ctggaaatca ctcacacttc tgggatgcct tcagagcagc
     5881 cctctatccc ttcagctccc ctgggatgtg actcgacctc ccgtcactcc ccagactgcc
     5941 tctgccaagt ccgaaagtgg aggcatcctt gcgagcaagt aggcgggtcc agggtggcgc
     6001 atgtcactca tcgaaagtgg aggcgtcctt gcgagcaagc aggcgggtcc agggtggcgt
     6061 gtcactcatc cttttttctg gctaccaaag gtgcagataa ttaatgacta caaagaccat
     6121 gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa gaagaagctg
     6181 gatcttagca acgtccagtc caagtgtggc tcaaaggata atatcaaaca cgtcccggga
     6241 ggcggcagtg tgcaaatagt ctacaaacca gttgacctga gcaaggtgac ctccaagtgt
     6301 ggctcattag gcaacatcca tcataaacca ggaggtggcc aggtggaagt aaaatctgag
     6361 aagcttgact tcaaggacag agtccagtcg aagattgggt ccctggacaa tatcacccac
     6421 gtccctggcg gaggaaatat aaaggtaaag ggggtagggt gggttggatg ctgcccttgg
     6481 gtatatgggc attaatcaag ttgagtggac aaaggctggt ccagttccca gaggaggaaa
     6541 acagaggctt ctgtgttgac tggctggatg tgggccctca gcagcatcca gtgggtctcc
     6601 actgcctgtc tcaatcacct ggagctttag cacgtttcac acctgggccc caacctggag
     6661 aggctgacca atgggtctca ggggcagctc ggttgctgga gtttttgttt ttatttattt
     6721 ttatgtattt aaggcagggt ctctgtatta gtccattctc acactgctaa taaagacata
     6781 cccaagactg ggtaatttat aaaggaaaga ggtttaatgg actcacagtt ccatatggct
     6841 ggggaggcct caaaatcatg gcggaaggca aaggagaagc aaaggcatgt cttacatggc
     6901 aacaggcaag agagcgtgtg caggggaact cccatttata aaaccatcag acctcatgag
     6961 atttattcac tatcatgaga acagcatggg aaagacccgc ccccatgatt cagttacctc
     7021 ccactgggtc cctcccatga cacatggaat tatgggagct acaattcaag atgagatttg
     7081 ggtggggaca cagccaaacc atatcagtct ccctctgtca tccaggctgg agtgcactgg
     7141 catgatctcg gctcactgca gcctctacct ccctgggtca ggtgatcttc ccacctcagc
     7201 ctcccaggta gctggaacta caggtacctg ccactatgcc tggctaaata ttttgtattt
     7261 cctgtggaga cgaggttttg ccacgttgcc caggctggtc ttgaactcct gaggtcaagc
     7321 aatatgccca cctcggcctc ccaaggtgct gggattacag gtgtgagcca cagtgctcgg
     7381 cctaagtcac tgcagttttt agcggccgct cgagtctaga gggcccgttt aaacccgctg
     7441 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     7501 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     7561 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     7621 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     7681 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     7741 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     7801 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     7861 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     7921 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     7981 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     8041 aacaacactc a
//
