LOCUS       sequence-381056-        4452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged C-terminal domain of TDP-43 with 5 
            Methionines to tyrosine.
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-->Y_IDR
FEATURES             Location/Qualifiers
     source          1..4452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      527..1115
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1016..1035
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1269..1286
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1384..1401
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1385..1401
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1865..1884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1865..1883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1884..1908
                     /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             1969..1986
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1987..2007
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      2471..2518
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2741..2760)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        2827..2918
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2919..3728
                     /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(3811..4452,1..441)
                     /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(3830..3849)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
       61 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
      121 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
      181 gataccgaag atagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
      241 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
      301 ggcaatcagc tgttgccagt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
      361 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
      421 cgactggaaa gcgggcagtg actcatgacc aaaatccctt aacgtgagtt acgcgcgcgt
      481 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
      541 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
      601 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
      661 taccaaatac tgttcttcta gtgtagccgt agttagccca ccacttcaag aactctgtag
      721 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
      781 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
      841 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
      901 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
      961 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1021 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     1081 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     1141 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     1201 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     1261 ccgagcgcag cgagtcagtg agcgaggaag cggaaggcga gagtagggaa ctgccaggca
     1321 tcaaactaag cagaaggccc ctgacggatg gcctttttgc gtttctacaa actctttctg
     1381 tgttgtaaaa cgacggccag tcttaagctc gggccccctg ggcggttctg ataacgagta
     1441 atcgttaatc cgcaaataac gtaaaaaccc gcttcggcgg gtttttttat ggggggagtt
     1501 tagggaaaga gcatttgtca gaatatttaa gggcgcctgt cactttgctt gatatatgag
     1561 aattatttaa ccttataaat gagaaaaaag caacgcactt taaataagat acgttgcttt
     1621 ttcgattgat gaacacctat aattaaacta ttcatctatt atttatgatt ttttgtatat
     1681 acaatatttc tagtttgtta aagagaatta agaaaataaa tctcgaaaat aataaaggga
     1741 aaatcagttt ttgatatcaa aattatacat gtcaacgata atacaaaata taatacaaac
     1801 tataagatgt tatcagtatt tattatgcat ttagaataaa ttttgtgtcg cccttccgcg
     1861 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tctagaaata
     1921 attttgttta acttttgaag gagatatacc atgggcagcg acaagatcca ccatcaccac
     1981 catcacgaaa atctgtactt tcaaggtcat atgaaccgtc agctggagcg tagcggtcgt
     2041 ttcggtggca acccgggtgg cttcggcaac caaggtggct ttggtaacag ccgtggtggc
     2101 ggtgcgggtc tgggtaacaa ccagggtagc aactacggcg gtggctataa cttcggtgcg
     2161 tttagcatta acccggcgat gatggcggcg gcgcaggcgg cgctgcaaag cagctggggc
     2221 atgatgggta tgctggcgag ccagcaaaac caaagcggcc cgagcggtaa caaccagaac
     2281 caaggcaact accagcgtga accgaaccaa gcgtttggta gcggcaacaa cagctatagc
     2341 ggcagcaaca gcggtgcggc gattggttgg ggcagcgcga gcaacgcggg tagcggtagc
     2401 ggttttaacg gtggctttgg tagcagctac gacagcaaga gcagcggctg gggttattaa
     2461 ctcgagcccc ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgcc
     2521 cctgagacgc gtcaatcgag ttcgtaccta agggcgacac cccctaatta gcccgggcga
     2581 aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctggcagt tccctactct
     2641 cgcatgggga gtccccacac taccatcggc gctacggcgt ttcacttctg agttcggcat
     2701 ggggtcaggt gggaccaccg cgctactgcc gccaggcaaa caaggggtgt tatgagccat
     2761 attcaggtat aaatgggctc gcgataatgt tcagaattgg ttaattggtt gtaacactga
     2821 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2881 cctgataaat gcttcaataa tattgaaaaa ggaagaatat gagccatatt caacgggaaa
     2941 cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     3001 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc
     3061 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     3121 tcagactaaa ctggctgacg gaatttatgc cacttccgac catcaagcat tttatccgta
     3181 ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagcg ttccaggtat
     3241 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     3301 ggttgcactc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgcctcg
     3361 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     3421 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
     3481 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     3541 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     3601 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     3661 aatatggtat tgataatcct gatatgaata aattgcaatt tcatttgatg ctcgatgagt
     3721 ttttctaagc ggcgcgccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc
     3781 ccggaagaga gtcaattcag ggtggtgaat atgaaaccag taacgttata cgatgtcgca
     3841 gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt
     3901 tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac
     3961 cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt
     4021 ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg
     4081 ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg
     4141 gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac
     4201 caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc
     4261 tctgaccaga cacccatcaa cagtattatt ttctcccatg aggacggtac gcgactgggc
     4321 gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt
     4381 tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt
     4441 cagccgatag cg
//
