LOCUS       sequence-494486-        4452 bp    DNA     circular SYN 03-DEC-2025
DEFINITION  Expresses 6xHis-tagged C-terminal domain of TDP-43 with 13S-->E and 
            S317C..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4452)
  AUTHORS   Rizuan A, Shenoy J, Mohanty P, Dos Passos PMS, Mercado Ortiz JF, Bai
            L, Viswanathan R, Zaborowksy J, Wang SH, Johnson V, Mamede LD, Titus
            AR, Ayala YM, Ghirlando R, Mittal J, Fawzi NL
  TITLE     Structural details of helix-mediated multimerization of the 
            conserved region of TDP-43 C-terminal domain.
  JOURNAL   Nat Commun 16, 10528 (2025)
  PUBMED    39005345
REFERENCE   2  (bases 1 to 4452)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 3, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun 
            16, 10528 (2025)"
COMMENT     Sequence Label: TDP-43_CTD_13S-->E_S317C
FEATURES             Location/Qualifiers
     source          1..4452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      27..615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     516..535
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     769..786
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     884..901
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     885..901
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1365..1384
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1365..1383
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1384..1408
                     /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             1469..1486
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1487..1507
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      1971..2018
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2241..2260)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        2327..2418
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2419..3228
                     /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             3311..4393
                     /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(3330..3349)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
       61 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
      121 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
      181 gtgtagccgt agttagccca ccacttcaag aactctgtag caccgcctac atacctcgct
      241 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
      301 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
      361 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
      421 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
      481 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
      541 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
      601 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
      661 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
      721 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
      781 agcgaggaag cggaaggcga gagtagggaa ctgccaggca tcaaactaag cagaaggccc
      841 ctgacggatg gcctttttgc gtttctacaa actctttctg tgttgtaaaa cgacggccag
      901 tcttaagctc gggccccctg ggcggttctg ataacgagta atcgttaatc cgcaaataac
      961 gtaaaaaccc gcttcggcgg gtttttttat ggggggagtt tagggaaaga gcatttgtca
     1021 gaatatttaa gggcgcctgt cactttgctt gatatatgag aattatttaa ccttataaat
     1081 gagaaaaaag caacgcactt taaataagat acgttgcttt ttcgattgat gaacacctat
     1141 aattaaacta ttcatctatt atttatgatt ttttgtatat acaatatttc tagtttgtta
     1201 aagagaatta agaaaataaa tctcgaaaat aataaaggga aaatcagttt ttgatatcaa
     1261 aattatacat gtcaacgata atacaaaata taatacaaac tataagatgt tatcagtatt
     1321 tattatgcat ttagaataaa ttttgtgtcg cccttccgcg aaattaatac gactcactat
     1381 aggggaattg tgagcggata acaattcccc tctagaaata attttgttta acttttgaag
     1441 gagatatacc atgggcagcg acaagatcca ccatcaccac catcacgaaa atctgtactt
     1501 tcaaggtcat atgaaccgtc agctggagcg tagcggtcgt ttcggtggca acccgggtgg
     1561 cttcggcaac caaggtggct ttggtaacga gcgtggtggc ggtgcgggtc tgggtaacaa
     1621 ccagggtgaa aacatgggcg gtggcatgaa cttcggtgcg ttttgcatta acccggcgat
     1681 gatggcggcg gcgcaggcgg cgctgcaaag cagctggggc atgatgggta tgctggcgag
     1741 ccagcaaaac caaagcggcc cgagcggtaa caaccagaac caaggcaaca tgcagcgtga
     1801 gccgaaccaa gcgtttggtg aaggcaacaa cagctacagc ggcgagaacg agggtgcggc
     1861 gattggttgg ggcgaggcgg aaaacgcggg cgagggtgaa ggttttaacg gtggctttgg
     1921 cgaggaaatg gacagcaagg aagagggttg gggcatgtaa ctcgagcccc ctagcataac
     1981 cccttggggc ctctaaacgg gtcttgaggg gttttttgcc cctgagacgc gtcaatcgag
     2041 ttcgtaccta agggcgacac cccctaatta gcccgggcga aaggcccagt ctttcgactg
     2101 agcctttcgt tttatttgat gcctggcagt tccctactct cgcatgggga gtccccacac
     2161 taccatcggc gctacggcgt ttcacttctg agttcggcat ggggtcaggt gggaccaccg
     2221 cgctactgcc gccaggcaaa caaggggtgt tatgagccat attcaggtat aaatgggctc
     2281 gcgataatgt tcagaattgg ttaattggtt gtaacactga cccctatttg tttatttttc
     2341 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2401 tattgaaaaa ggaagaatat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat
     2461 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     2521 ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     2581 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     2641 gaatttatgc cacttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     2701 ctcaccactg cgatccccgg aaaaacagcg ttccaggtat tagaagaata tcctgattca
     2761 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcactc gattcctgtt
     2821 tgtaattgtc cttttaacag cgatcgcgta tttcgcctcg ctcaggcgca atcacgaatg
     2881 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     2941 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
     3001 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     3061 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     3121 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     3181 gatatgaata aattgcaatt tcatttgatg ctcgatgagt ttttctaagc ggcgcgccat
     3241 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
     3301 ggtggtgaat atgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
     3361 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
     3421 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
     3481 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
     3541 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
     3601 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
     3661 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
     3721 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
     3781 cagtattatt ttctcccatg aggacggtac gcgactgggc gtggagcatc tggtcgcatt
     3841 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
     3901 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
     3961 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
     4021 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
     4081 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga
     4141 agatagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
     4201 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
     4261 gctgttgcca gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
     4321 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     4381 aagcgggcag tgactcatga ccaaaatccc ttaacgtgag ttacgcgcgc gtcgttccac
     4441 tgagcgtcag ac
//
