LOCUS       sequence-381057-        4452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged C-terminal domain of TDP-43 with 5 
            Methionines to leucine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4452)
  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 4452)
  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_5M-->L_IDR
FEATURES             Location/Qualifiers
     source          1..4452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      568..1156
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1057..1076
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1310..1327
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1425..1442
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1426..1442
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1906..1925
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1906..1924
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1925..1949
                     /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)."
     CDS             2010..2027
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2028..2048
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      2512..2559
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2782..2801)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        2868..2959
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2960..3769
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             join(3852..4452,1..482)
                     /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     complement(3871..3890)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
       61 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
      121 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
      181 ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa gatagctcat
      241 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
      301 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgccag
      361 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
      421 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
      481 gactcatgac caaaatccct taacgtgagt tacgcgcgcg tcgttccact gagcgtcaga
      541 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
      601 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
      661 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
      721 agtgtagccg tagttagccc accacttcaa gaactctgta gcaccgccta catacctcgc
      781 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
      841 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
      901 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
      961 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1021 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1081 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1141 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1201 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1261 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     1321 gagcgaggaa gcggaaggcg agagtaggga actgccaggc atcaaactaa gcagaaggcc
     1381 cctgacggat ggcctttttg cgtttctaca aactctttct gtgttgtaaa acgacggcca
     1441 gtcttaagct cgggccccct gggcggttct gataacgagt aatcgttaat ccgcaaataa
     1501 cgtaaaaacc cgcttcggcg ggttttttta tggggggagt ttagggaaag agcatttgtc
     1561 agaatattta agggcgcctg tcactttgct tgatatatga gaattattta accttataaa
     1621 tgagaaaaaa gcaacgcact ttaaataaga tacgttgctt tttcgattga tgaacaccta
     1681 taattaaact attcatctat tatttatgat tttttgtata tacaatattt ctagtttgtt
     1741 aaagagaatt aagaaaataa atctcgaaaa taataaaggg aaaatcagtt tttgatatca
     1801 aaattataca tgtcaacgat aatacaaaat ataatacaaa ctataagatg ttatcagtat
     1861 ttattatgca tttagaataa attttgtgtc gcccttccgc gaaattaata cgactcacta
     1921 taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt aacttttgaa
     1981 ggagatatac catgggcagc gacaagatcc accatcacca ccatcacgaa aatctgtact
     2041 ttcaaggtca tatgaaccgt cagctggagc gtagcggtcg tttcggtggc aacccgggtg
     2101 gcttcggcaa ccaaggtggc tttggtaaca gccgtggtgg cggtgcgggt ctgggtaaca
     2161 accagggtag caacctgggc ggtggcctga acttcggtgc gtttagcatt aacccggcga
     2221 tgatggcggc ggcgcaggcg gcgctgcaaa gcagctgggg catgatgggt atgctggcga
     2281 gccagcaaaa ccaaagcggc ccgagcggta acaaccagaa ccaaggcaac ctgcagcgtg
     2341 aaccgaacca agcgtttggt agcggcaaca acagctacag cggcagcaac agcggtgcgg
     2401 cgattggttg gggcagcgcg agcaacgcgg gtagcggtag cggttttaac ggtggctttg
     2461 gtagcagcct ggacagcaag agcagcggct ggggtctgta actcgagccc cctagcataa
     2521 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc ccctgagacg cgtcaatcga
     2581 gttcgtacct aagggcgaca ccccctaatt agcccgggcg aaaggcccag tctttcgact
     2641 gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcatgggg agtccccaca
     2701 ctaccatcgg cgctacggcg tttcacttct gagttcggca tggggtcagg tgggaccacc
     2761 gcgctactgc cgccaggcaa acaaggggtg ttatgagcca tattcaggta taaatgggct
     2821 cgcgataatg ttcagaattg gttaattggt tgtaacactg acccctattt gtttattttt
     2881 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2941 atattgaaaa aggaagaata tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     3001 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     3061 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     3121 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     3181 ggaatttatg ccacttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     3241 actcaccact gcgatccccg gaaaaacagc gttccaggta ttagaagaat atcctgattc
     3301 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcact cgattcctgt
     3361 ttgtaattgt ccttttaaca gcgatcgcgt atttcgcctc gctcaggcgc aatcacgaat
     3421 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     3481 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     3541 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     3601 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     3661 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     3721 tgatatgaat aaattgcaat ttcatttgat gctcgatgag tttttctaag cggcgcgcca
     3781 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
     3841 gggtggtgaa tatgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
     3901 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
     3961 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
     4021 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
     4081 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
     4141 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
     4201 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
     4261 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
     4321 acagtattat tttctcccat gaggacggta cgcgactggg cgtggagcat ctggtcgcat
     4381 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
     4441 gtctggctgg ct
//
