LOCUS       sequence-189284-        4383 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses 6xHis-tagged C-terminal domain of TDP-43 with a deletion 
            of AA 321-343.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4383)
  AUTHORS   Conicella AE, Zerze GH, Mittal J, Fawzi NL
  TITLE     ALS Mutations Disrupt Phase Separation Mediated by alpha-Helical 
            Structure in the TDP-43 Low-Complexity C-Terminal Domain.
  JOURNAL   Structure. 2016 Sep 6;24(9):1537-49. doi: 10.1016/j.str.2016.07.007.
            Epub 2016 Aug 18.
  PUBMED    27545621
REFERENCE   2  (bases 1 to 4383)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.str.2016.07.007"; journalName: "Structure"; date: 
            "2016-09-6- 6"; volume: "24"; issue: "9"; pages: "1537-49"
COMMENT     Sequence Label: TDP-43_CTD_del321-343
FEATURES             Location/Qualifiers
     source          1..4383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             105..122
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             123..143
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      538..585
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(808..827)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        894..985
                     /gene="bla"
                     /label=AmpR promoter
     CDS             986..1795
                     /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             1878..2960
                     /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(1897..1916)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      3046..3634
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3535..3554
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3788..3805
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3903..3920
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3904..3920
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt ttgaaggaga tataccatgg gtagcgacaa gattcatcac caccatcatc
      121 atgagaactt atactttcaa ggccatatga accgccaact ggaacgttcc ggtcgtttcg
      181 gtggtaatcc gggtggtttc ggcaaccagg gtggctttgg caattcgcgt ggcggtggtg
      241 ccggcttggg caataaccaa ggttccaata tgggtggcgg tatgaacttt ggtgccttct
      301 ccatcaatcc gcagaaccag tcaggcccga gcggcaataa tcagaatcaa ggtaacatgc
      361 agcgtgaacc gaatcaagcg tttggtagcg gtaacaatag ctacagcggt agcaacagcg
      421 gtgcggcaat cggctggggt agcgcaagca atgcgggcag cggtagcggt ttcaacggcg
      481 gctttggtag cagcatggat agcaagagct ccggttgggg catgtaactc gagcccccta
      541 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgcccct gagacgcgtc
      601 aatcgagttc gtacctaagg gcgacacccc ctaattagcc cgggcgaaag gcccagtctt
      661 tcgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc atggggagtc
      721 cccacactac catcggcgct acggcgtttc acttctgagt tcggcatggg gtcaggtggg
      781 accaccgcgc tactgccgcc aggcaaacaa ggggtgttat gagccatatt caggtataaa
      841 tgggctcgcg ataatgttca gaattggtta attggttgta acactgaccc ctatttgttt
      901 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
      961 tcaataatat tgaaaaagga agaatatgag ccatattcaa cgggaaacgt cgaggccgcg
     1021 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     1081 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     1141 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     1201 gctgacggaa tttatgccac ttccgaccat caagcatttt atccgtactc ctgatgatgc
     1261 atggttactc accactgcga tccccggaaa aacagcgttc caggtattag aagaatatcc
     1321 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcactcgat
     1381 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgcctcgctc aggcgcaatc
     1441 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     1501 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     1561 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     1621 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     1681 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     1741 taatcctgat atgaataaat tgcaatttca tttgatgctc gatgagtttt tctaagcggc
     1801 gcgccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
     1861 aattcagggt ggtgaatatg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
     1921 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
     1981 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
     2041 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
     2101 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
     2161 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
     2221 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
     2281 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
     2341 ccatcaacag tattattttc tcccatgagg acggtacgcg actgggcgtg gagcatctgg
     2401 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc
     2461 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg
     2521 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg
     2581 agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc
     2641 gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg
     2701 ataccgaaga tagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc
     2761 tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg
     2821 gcaatcagct gttgccagtc tcactggtga aaagaaaaac caccctggcg cccaatacgc
     2881 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     2941 gactggaaag cgggcagtga ctcatgacca aaatccctta acgtgagtta cgcgcgcgtc
     3001 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3061 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3121 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3181 accaaatact gttcttctag tgtagccgta gttagcccac cacttcaaga actctgtagc
     3241 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3301 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3361 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3421 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3481 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3541 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3601 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3661 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     3721 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     3781 cgagcgcagc gagtcagtga gcgaggaagc ggaaggcgag agtagggaac tgccaggcat
     3841 caaactaagc agaaggcccc tgacggatgg cctttttgcg tttctacaaa ctctttctgt
     3901 gttgtaaaac gacggccagt cttaagctcg ggccccctgg gcggttctga taacgagtaa
     3961 tcgttaatcc gcaaataacg taaaaacccg cttcggcggg tttttttatg gggggagttt
     4021 agggaaagag catttgtcag aatatttaag ggcgcctgtc actttgcttg atatatgaga
     4081 attatttaac cttataaatg agaaaaaagc aacgcacttt aaataagata cgttgctttt
     4141 tcgattgatg aacacctata attaaactat tcatctatta tttatgattt tttgtatata
     4201 caatatttct agtttgttaa agagaattaa gaaaataaat ctcgaaaata ataaagggaa
     4261 aatcagtttt tgatatcaaa attatacatg tcaacgataa tacaaaatat aatacaaact
     4321 ataagatgtt atcagtattt attatgcatt tagaataaat tttgtgtcgc ccttccgcga
     4381 aat
//
