LOCUS       sequence-381053-        5739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged C-terminal domain of TDP-43 with 6F-->A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5739)
  AUTHORS   Mohanty P, Shenoy J, Rizuan A, Mercado-Ortiz JF, Fawzi NL, Mittal J
  TITLE     A synergy between site-specific and transient interactions drives 
            the phase separation of a disordered, low-complexity domain.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Aug 22;120(34):e2305625120. doi: 
            10.1073/pnas.2305625120. Epub 2023 Aug 14.
  PUBMED    37579155
REFERENCE   2  (bases 1 to 5739)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2023-08-22"; volume: "120(34):e2305625120. 
            doi"; pages: " 10.1073/pnas.2305625120. Epub 2023 Aug 14"
COMMENT     Sequence Label: TDP-43 _CTD_6F-->A
FEATURES             Location/Qualifiers
     source          1..5739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     26..48
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(42..233)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1042..2124)
                     /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     2086..2105
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2125..2202)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2408..2427
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2511..2530
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2511..2529
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2530..2554
                     /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             2569..2591
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2616..2633
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2634..2654
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3114..3131
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3184..3202)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3198..3245
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3282..3737
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3369..3388)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3579..3600
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3829..4644)
                     /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(3829..4644)
                     /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     4552..4571
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4636..4655
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4766..5354
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5255..5274
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5508..5525
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5540..5679
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5605..5624)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
       61 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
      121 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
      181 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
      241 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
      301 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
      361 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
      421 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
      481 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
      541 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
      601 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
      661 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat gccggcgata
      721 atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg
      781 gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag
      841 cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata
      901 aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg
      961 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
     1021 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     1081 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
     1141 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
     1201 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
     1261 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
     1321 gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
     1381 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
     1441 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
     1501 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
     1561 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
     1621 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
     1681 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
     1741 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
     1801 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
     1861 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
     1921 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
     1981 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
     2041 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
     2101 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
     2161 tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc
     2221 ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg
     2281 ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     2341 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     2401 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     2461 cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat taatacgact
     2521 cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
     2581 taagaaggag atataccatg ggctctgata aaattcatca tcatcatcat cacgaaaatc
     2641 tttattttca aggtcatatg aaccgtcaac tggagcgtag cggtcgtgcg ggtggcaacc
     2701 cgggtggcgc gggtaaccaa ggtggcgcgg gtaacagccg tggtggcggt gcgggtctgg
     2761 gtaacaacca gggtagcaac atgggcggtg gcatgaactt cggtgcgttt agcattaacc
     2821 cggcgatgat ggcggcggcg caggcggcgc tgcaaagcag ctggggcatg atgggtatgc
     2881 tggcgagcca gcaaaaccaa agcggcccga gcggtaacaa ccagaaccaa ggcaacatgc
     2941 agcgtgaacc gaaccaagcg gcgggtagcg gcaacaacag ctacagcggc agcaacagcg
     3001 gtgcggcgat tggttggggc agcgcgagca acgcgggtag cggtagcggt gcgaacggtg
     3061 gcgcgggtag cagcatggac agcaagagca gcggctgggg catgtaactc gagcaccacc
     3121 accaccacca ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     3181 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     3241 ttttgctgaa aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc
     3301 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3361 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3421 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3481 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3541 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3601 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3661 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3721 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3781 ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc
     3841 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
     3901 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     3961 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     4021 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
     4081 gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc
     4141 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
     4201 acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
     4261 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     4321 ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     4381 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     4441 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     4501 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     4561 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga
     4621 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
     4681 ttttattgtt catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4741 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4801 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4861 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     4921 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4981 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5041 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5101 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5161 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5221 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5281 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5341 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5401 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5461 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     5521 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     5581 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     5641 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     5701 gctgacgcgc cctgacgggc ttgtctgctc ccggcatcc
//
