LOCUS       sequence-372451-        3766 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of 6xHis-tagged P301S tau of 0N4R isoform with a 
            C-terminal HiBiT tag for bimolecular luminescence with LgBiT..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3766)
  AUTHORS   Tuck BJ, Miller LVC, Katsinelos T, Smith AE, Wilson EL, Keeling S, 
            Cheng S, Vaysburd MJ, Knox C, Tredgett L, Metzakopian E, James LC, 
            McEwan WA
  TITLE     Cholesterol determines the cytosolic entry and seeded aggregation of
            tau.
  JOURNAL   Cell Rep. 2022 May 3;39(5):110776. doi: 
            10.1016/j.celrep.2022.110776.
  PUBMED    35508140
REFERENCE   2  (bases 1 to 3766)
  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: "Cell Rep.";
            date: "2022-05-3"; volume: "39(5):110776. doi"; pages: " 
            10.1016/j.celrep.2022.110776"
COMMENT     Sequence Label: pRK172-6xHis-0N4R-P301S-tau-HiBiT
FEATURES             Location/Qualifiers
     source          1..3766
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..37)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        105..209
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(210..278,279..1070)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(428..447)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1241..1829
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1730..1749
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1983..2000
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2015..2154
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2080..2099)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2250..2272
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2325..2344
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2325..2343
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2375..2397
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2411..2428
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3679..3697)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3693..3740
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
       61 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
      121 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
      181 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
      241 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
      301 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
      361 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
      421 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
      481 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
      541 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
      601 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
      661 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
      721 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
      781 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
      841 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
      901 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
      961 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     1021 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     1081 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     1141 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1201 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1261 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1321 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1381 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     1441 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     1501 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     1561 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     1621 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     1681 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     1741 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     1801 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     1861 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     1921 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     1981 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     2041 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     2101 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     2161 cacccgccaa cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     2221 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     2281 gtcatcaccg aaacgcgcga ggcaggatct cgatcccgcg aaattaatac gactcactat
     2341 agggagacca caacggtttc cctctagaaa taattttgtt taactttaag aaggagatat
     2401 acatatgatg catcaccatc accatcacgc tgagccccgc caggagttcg aagtgatgga
     2461 agatcacgct gggacgtacg ggttggggga caggaaagat caggggggct acaccatgca
     2521 ccaagaccaa gagggtgaca cggacgctgg cctgaaagct gaagaagcag gcattggaga
     2581 cacccccagc ctggaagacg aagctgctgg tcacgtgacc caagctcgca tggtcagtaa
     2641 aagcaaagac gggactggaa gcgatgacaa aaaagccaag ggggctgatg gtaaaacgaa
     2701 gatcgccaca ccgcggggag cagcccctcc aggccagaag ggccaggcca acgccaccag
     2761 gattccagca aaaaccccgc ccgctccaaa gacaccaccc agctctggtg aacctccaaa
     2821 atcaggggat cgcagcggct acagcagccc cggctcccca ggcactcccg gcagccgctc
     2881 ccgcaccccg tcccttccaa ccccacccac ccgggagccc aagaaggtgg cagtggtccg
     2941 tactccaccc aagtcgccgt cttccgccaa gagccgcctg cagacagccc ccgtgcccat
     3001 gccagacctg aagaatgtca agtccaagat cggctccact gagaacctga agcaccagcc
     3061 gggaggcggg aaggtgcaga taattaataa gaagctggat cttagcaacg tccagtccaa
     3121 gtgtggctca aaggataata tcaaacacgt cagtggaggc ggcagtgtgc aaatagtcta
     3181 caaaccagtt gacctgagca aggtgacctc caagtgtggc tcattaggca acatccatca
     3241 taaaccagga ggtggccagg tggaagtaaa atctgagaag cttgacttca aggacagagt
     3301 ccagtcgaag attgggtccc tggacaatat cacccacgtc cctggcggag gaaataaaaa
     3361 gattgaaacc cacaagctga ccttccgcga gaacgccaaa gccaagacag accacggggc
     3421 ggagatcgtg tacaagtcgc cagtggtgtc tggggacacg tctccacggc atctcagcaa
     3481 tgtctcctcc accggcagca tcgacatggt agactcgccc cagctcgcca cgctagctga
     3541 cgaggtgtct gcctccctgg ccaagcaggg tttgggttca ggttcagtga gcggctggcg
     3601 cctgttcaag aagatcagct aggaattccg atccggctgc taacaaagcc cgaaaggaag
     3661 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     3721 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggata
//
