LOCUS       sequence-381055-        4452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged C-terminal domain of TDP-43 with 5 Alanines 
            to methionine.
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_5A-->M_IDR
FEATURES             Location/Qualifiers
     source          1..4452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      171..759
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     660..679
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     913..930
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1028..1045
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1029..1045
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1509..1528
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1509..1527
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1528..1552
                     /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             1613..1630
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1631..1651
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(1758..1775)
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     terminator      2115..2162
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2385..2404)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        2471..2562
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2563..3372
                     /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(3455..4452,1..85)
                     /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(3474..3493)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
       61 ttcccgactg gaaagcgggc agtgactcat gaccaaaatc ccttaacgtg agttacgcgc
      121 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
      181 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
      241 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
      301 cagataccaa atactgttct tctagtgtag ccgtagttag cccaccactt caagaactct
      361 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
      421 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
      481 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
      541 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
      601 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
      661 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
      721 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
      781 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
      841 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
      901 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag gcgagagtag ggaactgcca
      961 ggcatcaaac taagcagaag gcccctgacg gatggccttt ttgcgtttct acaaactctt
     1021 tctgtgttgt aaaacgacgg ccagtcttaa gctcgggccc cctgggcggt tctgataacg
     1081 agtaatcgtt aatccgcaaa taacgtaaaa acccgcttcg gcgggttttt ttatgggggg
     1141 agtttaggga aagagcattt gtcagaatat ttaagggcgc ctgtcacttt gcttgatata
     1201 tgagaattat ttaaccttat aaatgagaaa aaagcaacgc actttaaata agatacgttg
     1261 ctttttcgat tgatgaacac ctataattaa actattcatc tattatttat gattttttgt
     1321 atatacaata tttctagttt gttaaagaga attaagaaaa taaatctcga aaataataaa
     1381 gggaaaatca gtttttgata tcaaaattat acatgtcaac gataatacaa aatataatac
     1441 aaactataag atgttatcag tatttattat gcatttagaa taaattttgt gtcgcccttc
     1501 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga
     1561 aataattttg tttaactttt gaaggagata taccatgggc agcgacaaga tccaccatca
     1621 ccaccatcac gaaaatctgt actttcaagg tcatatgaac cgtcagctgg agcgtagcgg
     1681 tcgtttcggt ggcaacccgg gtggcttcgg caaccaaggt ggctttggta acagccgtgg
     1741 tggcggtatg ggtctgggca acaaccaggg cagcaacatg ggcggtggca tgaacttcgg
     1801 tgcgtttagc attaacccgg cgatgatggc ggcggcgcag gcggcgctgc aaagcagctg
     1861 gggcatgatg ggtatgctgg cgagccagca aaaccaaagc ggcccgagcg gtaacaacca
     1921 gaaccaaggc aacatgcagc gtgaaccgaa ccaaatgttt ggtagcggca acaacagcta
     1981 cagcggcagc aacagcggta tggcgattgg ttggggcagc atgagcaaca tgggtagcgg
     2041 cagcggtttt aacggtggct ttggtagcag catggacagc aagagcagcg gctggggcat
     2101 gtaactcgag ccccctagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     2161 tgcccctgag acgcgtcaat cgagttcgta cctaagggcg acacccccta attagcccgg
     2221 gcgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgcctgg cagttcccta
     2281 ctctcgcatg gggagtcccc acactaccat cggcgctacg gcgtttcact tctgagttcg
     2341 gcatggggtc aggtgggacc accgcgctac tgccgccagg caaacaaggg gtgttatgag
     2401 ccatattcag gtataaatgg gctcgcgata atgttcagaa ttggttaatt ggttgtaaca
     2461 ctgaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2521 taaccctgat aaatgcttca ataatattga aaaaggaaga atatgagcca tattcaacgg
     2581 gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     2641 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat
     2701 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     2761 atggtcagac taaactggct gacggaattt atgccacttc cgaccatcaa gcattttatc
     2821 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcgttccag
     2881 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     2941 cgccggttgc actcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgc
     3001 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     3061 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     3121 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     3181 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     3241 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     3301 caaaaatatg gtattgataa tcctgatatg aataaattgc aatttcattt gatgctcgat
     3361 gagtttttct aagcggcgcg ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata
     3421 gcgcccggaa gagagtcaat tcagggtggt gaatatgaaa ccagtaacgt tatacgatgt
     3481 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
     3541 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
     3601 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
     3661 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
     3721 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
     3781 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
     3841 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
     3901 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaggacg gtacgcgact
     3961 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
     4021 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
     4081 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
     4141 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
     4201 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
     4261 ggtagtggga tacgacgata ccgaagatag ctcatgttat atcccgccgt taaccaccat
     4321 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
     4381 gggccaggcg gtgaagggca atcagctgtt gccagtctca ctggtgaaaa gaaaaaccac
     4441 cctggcgccc aa
//
