LOCUS       sequence-401580-        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 S305I
FEATURES             Location/Qualifiers
     source          1..8051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(118..135)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(289..877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(369..388)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1048..1839,1840..1908))
                     /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     1671..1690
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1909..2013)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2088..2107)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2279..2658
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2659..2862
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2813..2833
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2907..2926
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2907..2925
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4346..4411
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5293..5304
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(5686..5703)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5692..5916
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5962..6390
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6049..6068)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6259..6280
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6399..6419)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6404..6733
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6584..6719
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6800..7594
                     /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
                     VLFVKTDLSGALNELQDEAAWLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6854..6873)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7464..7483
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7768..7889
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7805..7824)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7859..7878
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7938..7954)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7938..7954)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7951..7973)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7962..7978
                     /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(7986..7992,7993..8010,8011..8016))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(8031..8051,1)
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
       61 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
      121 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
      181 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
      241 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
      301 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
      361 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
      421 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
      481 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
      541 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
      601 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
      661 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
      721 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
      781 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
      841 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
      901 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
      961 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1021 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     1081 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     1141 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     1201 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     1261 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     1321 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     1381 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     1441 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     1501 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     1561 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     1621 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     1681 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     1741 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     1801 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     1861 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     1921 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     1981 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     2041 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     2101 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     2161 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     2221 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
     2281 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     2341 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     2401 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     2461 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     2521 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     2581 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     2641 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     2701 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     2761 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     2821 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa
     2881 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagctggc
     2941 tagcgtttaa acttaagctt ggtaccgagc tcggatccac tagtccagtg tggtggaatt
     3001 cccaggctga tggggcctcc agggcagtgg tttccaccca gcattctcaa aagagccact
     3061 gaggtctcag tgccattttc aagatttcgg aagcggcctg ggcacggctg gtccttcact
     3121 gggatcacca cttggcaatt atttacacct gagacgaata aaaaccagag tgctgagatt
     3181 acaggcatgg tggcttacgc ttgtaatcgg ctttgggaag ccgaggtggg ctgattgctt
     3241 gagcccagga gtttcaaact atcctggaca acatagcatg acctcgtctc tacaaaaaat
     3301 acaaaaaatt tgccaggtgt ggtggcatgt gcctgtggtc ccagctactt gggaggctga
     3361 agtaggagaa tcccctgagc cctgggaagt cgaggctgca ctgagccgtg atgatgtcac
     3421 tgcactccag cctgggtgac aaagtgagac cctatctcac aaagaaaaaa aacaaaacaa
     3481 aaaacccaaa gcacactgtt tccactgttt ccagagttcc tgagaggaaa ggtcaccggg
     3541 tgaggaagac gttctcactg atctggcaga gaaaatgtcc agtttttcca actccctaaa
     3601 ccatggtttt ctatttcaca gccccccctt gcagggacag agccctcgta cagaagggac
     3661 accccacatt tgtcttcccc acaaagcggc ctgtgtcctg cctacggggt cagggcttct
     3721 caaacctggc tgtgtgtcag aatcaccagg ggaacttttc aaaactagag agactgaagc
     3781 cagactccta gattctaatt ctaggtcagg gctaggggct gagattgtaa aaatccacag
     3841 gtgattctga tgcccggcag gcttgagaac agccgcaggg agttctctgg gaatgtgccg
     3901 gtgggtctag ccaggtgtga gtggagatgc cggggaactt cctattactc actcgtcagt
     3961 gtggccgaac acatttttca cttgacctca ggctggtgaa cgctcccctc tggggttcag
     4021 gcctcacgat gccatccttt tgtgaagtga ggacctgcaa tcccagcttc gtaaagcccg
     4081 ctggaaatca ctcacacttc tgggatgcct tcagagcagc cctctatccc ttcagctccc
     4141 ctgggatgtg actcgacctc ccgtcactcc ccagactgcc tctgccaagt ccgaaagtgg
     4201 aggcatcctt gcgagcaagt aggcgggtcc agggtggcgc atgtcactca tcgaaagtgg
     4261 aggcgtcctt gcgagcaagc aggcgggtcc agggtggcgt gtcactcatc cttttttctg
     4321 gctaccaaag gtgcagataa ttaatgacta caaagaccat gacggtgatt ataaagatca
     4381 tgacatcgat tacaaggatg acgatgacaa gaagaagctg gatcttagca acgtccagtc
     4441 caagtgtggc tcaaaggata atatcaaaca cgtcccggga ggcggcattg tgcaaatagt
     4501 ctacaaacca gttgacctga gcaaggtgac ctccaagtgt ggctcattag gcaacatcca
     4561 tcataaacca ggaggtggcc aggtggaagt aaaatctgag aagcttgact tcaaggacag
     4621 agtccagtcg aagattgggt ccctggacaa tatcacccac gtccctggcg gaggaaataa
     4681 aaaggtaaag ggggtagggt gggttggatg ctgcccttgg gtatatgggc attaatcaag
     4741 ttgagtggac aaaggctggt ccagttccca gaggaggaaa acagaggctt ctgtgttgac
     4801 tggctggatg tgggccctca gcagcatcca gtgggtctcc actgcctgtc tcaatcacct
     4861 ggagctttag cacgtttcac acctgggccc caacctggag aggctgacca atgggtctca
     4921 ggggcagctc ggttgctgga gtttttgttt ttatttattt ttatgtattt aaggcagggt
     4981 ctctgtatta gtccattctc acactgctaa taaagacata cccaagactg ggtaatttat
     5041 aaaggaaaga ggtttaatgg actcacagtt ccatatggct ggggaggcct caaaatcatg
     5101 gcggaaggca aaggagaagc aaaggcatgt cttacatggc aacaggcaag agagcgtgtg
     5161 caggggaact cccatttata aaaccatcag acctcatgag atttattcac tatcatgaga
     5221 acagcatggg aaagacccgc ccccatgatt cagttacctc ccactgggtc cctcccatga
     5281 cacatggaat tatgggagct acaattcaag atgagatttg ggtggggaca cagccaaacc
     5341 atatcagtct ccctctgtca tccaggctgg agtgcactgg catgatctcg gctcactgca
     5401 gcctctacct ccctgggtca ggtgatcttc ccacctcagc ctcccaggta gctggaacta
     5461 caggtacctg ccactatgcc tggctaaata ttttgtattt cctgtggaga cgaggttttg
     5521 ccacgttgcc caggctggtc ttgaactcct gaggtcaagc aatatgccca cctcggcctc
     5581 ccaaggtgct gggattacag gtgtgagcca cagtgctcgg cctaagtcac tgcagttttt
     5641 agcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     5701 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     5761 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     5821 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     5881 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     5941 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     6001 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     6061 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     6121 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     6181 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     6241 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     6301 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     6361 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     6421 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     6481 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     6541 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     6601 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     6661 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     6721 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     6781 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     6841 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     6901 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     6961 cggtgccctg aatgaactgc aggacgaggc agcgtggcta tcgtggctgg ccacgacggg
     7021 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     7081 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     7141 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     7201 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     7261 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     7321 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     7381 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     7441 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     7501 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     7561 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     7621 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     7681 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     7741 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     7801 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     7861 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     7921 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     7981 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     8041 taactcacat t
//
