LOCUS       sequence-381058-        4452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged C-terminal domain of TDP-43 with 5 
            Methionines to Alanine.
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-->A_IDR
FEATURES             Location/Qualifiers
     source          1..4452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      471..1059
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     960..979
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1213..1230
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1328..1345
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1329..1345
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1809..1828
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1809..1827
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1828..1852
                     /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             1913..1930
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1931..1951
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      2415..2462
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2685..2704)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        2771..2862
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2863..3672
                     /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(3755..4452,1..385)
                     /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(3774..3793)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc
       61 aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata
      121 cgacgatacc gaagatagct catgttatat cccgccgtta accaccatca aacaggattt
      181 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt
      241 gaagggcaat cagctgttgc cagtctcact ggtgaaaaga aaaaccaccc tggcgcccaa
      301 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
      361 ttcccgactg gaaagcgggc agtgactcat gaccaaaatc ccttaacgtg agttacgcgc
      421 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
      481 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
      541 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
      601 cagataccaa atactgttct tctagtgtag ccgtagttag cccaccactt caagaactct
      661 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
      721 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
      781 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
      841 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
      901 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
      961 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     1021 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     1081 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     1141 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     1201 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag gcgagagtag ggaactgcca
     1261 ggcatcaaac taagcagaag gcccctgacg gatggccttt ttgcgtttct acaaactctt
     1321 tctgtgttgt aaaacgacgg ccagtcttaa gctcgggccc cctgggcggt tctgataacg
     1381 agtaatcgtt aatccgcaaa taacgtaaaa acccgcttcg gcgggttttt ttatgggggg
     1441 agtttaggga aagagcattt gtcagaatat ttaagggcgc ctgtcacttt gcttgatata
     1501 tgagaattat ttaaccttat aaatgagaaa aaagcaacgc actttaaata agatacgttg
     1561 ctttttcgat tgatgaacac ctataattaa actattcatc tattatttat gattttttgt
     1621 atatacaata tttctagttt gttaaagaga attaagaaaa taaatctcga aaataataaa
     1681 gggaaaatca gtttttgata tcaaaattat acatgtcaac gataatacaa aatataatac
     1741 aaactataag atgttatcag tatttattat gcatttagaa taaattttgt gtcgcccttc
     1801 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga
     1861 aataattttg tttaactttt gaaggagata taccatgggc agcgacaaga tccaccatca
     1921 ccaccatcac gaaaatctgt actttcaagg tcatatgaac cgtcagctgg agcgtagcgg
     1981 tcgtttcggt ggcaacccgg gtggcttcgg caaccaaggt ggctttggta acagccgtgg
     2041 tggcggtgcg ggtctgggta acaaccaggg tagcaacgcg ggcggtggcg cgaactttgg
     2101 tgcgtttagc attaacccgg cgatgatggc ggcggcgcag gcggcgctgc aaagcagctg
     2161 gggcatgatg ggtatgctgg cgagccagca aaaccaaagc ggcccgagcg gtaacaacca
     2221 gaaccaaggc aacgcgcagc gtgaaccgaa ccaagcgttt ggtagcggca acaacagcta
     2281 cagcggcagc aacagcggtg cggcgattgg ttggggcagc gcgagcaacg cgggtagcgg
     2341 tagcggtttt aacggtggct ttggtagcag cgcggacagc aagagcagcg gttggggtgc
     2401 gtaactcgag ccccctagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     2461 tgcccctgag acgcgtcaat cgagttcgta cctaagggcg acacccccta attagcccgg
     2521 gcgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgcctgg cagttcccta
     2581 ctctcgcatg gggagtcccc acactaccat cggcgctacg gcgtttcact tctgagttcg
     2641 gcatggggtc aggtgggacc accgcgctac tgccgccagg caaacaaggg gtgttatgag
     2701 ccatattcag gtataaatgg gctcgcgata atgttcagaa ttggttaatt ggttgtaaca
     2761 ctgaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2821 taaccctgat aaatgcttca ataatattga aaaaggaaga atatgagcca tattcaacgg
     2881 gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     2941 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat
     3001 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     3061 atggtcagac taaactggct gacggaattt atgccacttc cgaccatcaa gcattttatc
     3121 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcgttccag
     3181 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     3241 cgccggttgc actcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgc
     3301 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     3361 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     3421 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     3481 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     3541 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     3601 caaaaatatg gtattgataa tcctgatatg aataaattgc aatttcattt gatgctcgat
     3661 gagtttttct aagcggcgcg ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata
     3721 gcgcccggaa gagagtcaat tcagggtggt gaatatgaaa ccagtaacgt tatacgatgt
     3781 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
     3841 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
     3901 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
     3961 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
     4021 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
     4081 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
     4141 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
     4201 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaggacg gtacgcgact
     4261 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
     4321 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
     4381 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
     4441 catgcaaatg ct
//
