LOCUS       sequence-209739-        6738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Tau with a mutated cysteine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6738)
  TITLE     Bacterial expression plasmids and lentivirus expression vectors for 
            tau and alpha-synuclein
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6738)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pET29b Tau Cys
FEATURES             Location/Qualifiers
     source          1..6738
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     213..232
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        213..231
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    232..256
                     /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)."
     RBS             271..293
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1811..1828
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1881..1899)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1895..1942
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1979..2434
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2066..2085)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2276..2297
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2526..3341)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2526..3341)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3249..3268
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3333..3352
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3463..4051
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3952..3971
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4205..4222
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4237..4376
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4302..4321)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4462..4484
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4478..4669)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5478..6560)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6522..6541
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6561..6638)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
       61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
      181 gaggatcgag atcgatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg
      241 agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacat
      301 atggctgagc cccgccagga gttcgaagtg atggaagatc acgctgggac gtacgggttg
      361 ggggacagga aagatcaggg gggctacacc atgcaccaag accaagaggg tgacacggac
      421 gctggcctga aagaatctcc cctgcagacc cccactgagg acggatctga ggaaccgggc
      481 tctgaaacct ctgatgctaa gagcactcca acagcggaag atgtgacagc acccttagtg
      541 gatgagggag ctcccggcaa gcaggctgcc gcgcagcccc acacggagat cccagaagga
      601 accacagctg aagaagcagg cattggagac acccccagcc tggaagacga agctgctggt
      661 cacgtgaccc aagctcgcat ggtcagtaaa agcaaagacg ggactggaag cgatgacaaa
      721 aaagccaagg gggctgatgg taaaacgaag atcgccacac cgcggggagc agcccctcca
      781 ggccagaagg gccaggccaa cgccaccagg attccagcaa aaaccccgcc cgctccaaag
      841 acaccaccca gctctggtga acctccaaaa tcaggggatc gcagcggcta cagcagcccc
      901 ggctccccag gcactcccgg cagccgctcc cgcaccccgt cccttccaac cccacccacc
      961 cgggagccca agaaggtggc agtggtccgt actccaccca agtcgccgtc ttccgccaag
     1021 agccgcctgc agacagcccc cgtgcccatg ccagacctga agaatgtcaa gtccaagtgc
     1081 ggctccactg agaacctgaa gcaccagccg ggaggcggga aggtgcagat aattaataag
     1141 aagctggatc ttagcaacgt ccagtccaag gcgggctcaa aggataatat caaacacgtc
     1201 ccgggaggcg gcagtgtgca aatagtctac aaaccagttg acctgagcaa ggtgacctcc
     1261 aaggcaggct cattaggcaa catccatcat aaaccaggag gtggccaggt ggaagtaaaa
     1321 tctgagaagc ttgacttcaa ggacagagtc cagtcgaaga ttgggtccct ggacaatatc
     1381 acccacgtcc ctggcggagg aaataaaaag attgaaaccc acaagctgac cttccgcgag
     1441 aacgccaaag ccaagacaga ccacggggcg gagatcgtgt acaagtcgcc agtggtgtct
     1501 ggggacacgt ctccacggca tctcagcaat gtctcctcca ccggcagcat cgacatggta
     1561 gactcgcccc agctcgccac gctagctgac gaggtgtctg cctccctggc caagcagggt
     1621 ttgtgatcag gcccctgggg cggtcaataa ttgtggagag gagagaatga gagagtgtgg
     1681 aaaaaaaaag aataatgacc cggcccccgc cctctgcccc cagctgctcc tcgcagttcg
     1741 ggaattagct aagaaggaga tataggatcc gaattcgagc tccgtcgaca agcttgcggc
     1801 cgcactcgag caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga
     1861 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     1921 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
     1981 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2041 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2101 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2161 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2221 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2281 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattattt tgatttataa
     2341 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2401 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     2461 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt
     2521 aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     2581 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     2641 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     2701 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     2761 acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca
     2821 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     2881 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     2941 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     3001 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     3061 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     3121 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     3181 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     3241 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     3301 cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     3361 tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg agttttcgtt
     3421 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3481 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3541 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3601 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3661 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3721 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3781 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3841 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3901 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3961 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4021 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4081 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     4141 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     4201 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     4261 gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc
     4321 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     4381 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4441 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4501 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     4561 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     4621 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     4681 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     4741 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     4801 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     4861 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     4921 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     4981 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     5041 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     5101 tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg
     5161 cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt
     5221 gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt
     5281 cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga
     5341 agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg
     5401 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     5461 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5521 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     5581 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     5641 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     5701 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg
     5761 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     5821 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     5881 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     5941 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     6001 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     6061 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     6121 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     6181 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     6241 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     6301 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     6361 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     6421 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     6481 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     6541 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     6601 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     6661 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     6721 cgcaaggaat ggtgcatg
//
