LOCUS       sequence-497374-        4594 bp    DNA     circular SYN 14-JAN-2026
DEFINITION  Recombinant protein synthesis of I277P/I308P mutatant of human 2N4R 
            Tau.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4594)
  TITLE     Efficient Tag-less Purification of Human Tau Proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4594)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 14, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pT7II-2N4Rtau I277P/I308P
FEATURES             Location/Qualifiers
     source          1..4594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(16..34)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        102..206
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(207..275,276..1067)
                     /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(425..444)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1238..1826
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1727..1746
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1980..1997
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2012..2151
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2077..2096)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2237..2259
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2495..2514
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2598..2617
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2598..2616
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2648..2670
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     rep_origin      complement(4070..4583)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4265..4286)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4477..4496
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
       61 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      121 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      181 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      241 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      301 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      361 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
      421 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
      481 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
      541 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
      601 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
      661 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
      721 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
      781 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
      841 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
      901 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
      961 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1021 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1081 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1141 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1201 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1261 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1321 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1381 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1441 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1501 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     1561 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     1621 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     1681 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     1741 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     1801 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     1861 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     1921 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     1981 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     2041 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     2101 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     2161 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     2221 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     2281 cgcgcgaggc agtcgatcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     2341 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     2401 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     2461 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     2521 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     2581 tctcgatccc gcgaaattaa tacgactcac tatagggaga ccacaacggt ttccctctag
     2641 aaataatttt gtttaacttt aagaaggaga tatacatatg gctgagcccc gccaggagtt
     2701 cgaagtgatg gaagatcacg ctgggacgta cgggttgggg gacaggaaag atcagggggg
     2761 ctacaccatg caccaagacc aagagggtga cacggacgct ggcctgaaag aatctcccct
     2821 gcagaccccc actgaggacg gatctgagga accgggctct gaaacctctg atgctaagag
     2881 cactccaaca gcggaagatg tgacagcacc cttagtggat gagggagctc ccggcaagca
     2941 ggctgccgcg cagccccaca cggagatccc agaaggaacc acagctgaag aagcaggcat
     3001 tggagacacc cccagcctgg aagacgaagc tgctggtcac gtgacccaag ctcgcatggt
     3061 cagtaaaagc aaagacggga ctggaagcga tgacaaaaaa gccaaggggg ctgatggtaa
     3121 aacgaagatc gccacaccgc ggggagcagc ccctccaggc cagaagggcc aggccaacgc
     3181 caccaggatt ccagcaaaaa ccccgcccgc tccaaagaca ccacccagct ctggtgaacc
     3241 tccaaaatca ggggatcgca gcggctacag cagccccggc tccccaggca ctcccggcag
     3301 ccgctcccgc accccgtccc ttccaacccc acccacccgg gagcccaaga aggtggcagt
     3361 ggtccgtact ccacccaagt cgccgtcttc cgccaagagc cgcctgcaga cagcccccgt
     3421 gcccatgcca gacctgaaga atgtcaagtc caagatcggc tccactgaga acctgaagca
     3481 ccagccggga ggcgggaagg tgcagccaat taataagaag ctggatctta gcaacgtcca
     3541 gtccaagtgt ggctcaaagg ataatatcaa acacgtcccg ggaggcggca gtgtgcaacc
     3601 agtctacaaa ccagttgacc tgagcaaggt gacctccaag tgtggctcat taggcaacat
     3661 ccatcataaa ccaggaggtg gccaggtgga agtaaaatct gagaagcttg acttcaagga
     3721 cagagtccag tcgaagattg ggtccctgga caatatcacc cacgtccctg gcggaggaaa
     3781 taaaaagatt gaaacccaca agctgacctt ccgcgagaac gccaaagcca agacagacca
     3841 cggggcggag atcgtgtaca agtcgccagt ggtgtctggg gacacgtctc cacggcatct
     3901 cagcaatgtc tcctccaccg gcagcatcga catggtagac tcgccccagc tcgccacgct
     3961 agctgacgag gtgtctgcct ccctggccaa gcagggtttg taagaattca agcttctcga
     4021 ggatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccccggttga
     4081 taatcagaaa agccccaaaa acaggaagat tgtataagca aatatttaaa ttgtaagcgt
     4141 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     4201 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     4261 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     4321 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     4381 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     4441 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     4501 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     4561 taatgcgccg ctacagggcg cgtggggatc aatt
//
