LOCUS       sequence-189272-        4452 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses 6xHis-tagged C-terminal domain of TDP-43 with A321V.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4452)
  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 4452)
  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_A321V
FEATURES             Location/Qualifiers
     source          1..4452
                     /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      607..654
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(877..896)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        963..1054
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1055..1864
                     /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             1947..3029
                     /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(1966..1985)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      3115..3703
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3604..3623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3857..3874
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3972..3989
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3973..3989
                     /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 ggtaatgatg gcggctgcac aagcggcgct gcagtctagc tggggtatga
      361 tgggcatgct ggcgagccag cagaaccagt caggcccgag cggcaataat cagaatcaag
      421 gtaacatgca gcgtgaaccg aatcaagcgt ttggtagcgg taacaatagc tacagcggta
      481 gcaacagcgg tgcggcaatc ggctggggta gcgcaagcaa tgcgggcagc ggtagcggtt
      541 tcaacggcgg ctttggtagc agcatggata gcaagagctc cggttggggc atgtaactcg
      601 agccccctag cataacccct tggggcctct aaacgggtct tgaggggttt tttgcccctg
      661 agacgcgtca atcgagttcg tacctaaggg cgacaccccc taattagccc gggcgaaagg
      721 cccagtcttt cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgca
      781 tggggagtcc ccacactacc atcggcgcta cggcgtttca cttctgagtt cggcatgggg
      841 tcaggtggga ccaccgcgct actgccgcca ggcaaacaag gggtgttatg agccatattc
      901 aggtataaat gggctcgcga taatgttcag aattggttaa ttggttgtaa cactgacccc
      961 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     1021 ataaatgctt caataatatt gaaaaaggaa gaatatgagc catattcaac gggaaacgtc
     1081 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     1141 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     1201 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     1261 actaaactgg ctgacggaat ttatgccact tccgaccatc aagcatttta tccgtactcc
     1321 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcgttcc aggtattaga
     1381 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     1441 gcactcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gcctcgctca
     1501 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     1561 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     1621 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     1681 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     1741 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     1801 tggtattgat aatcctgata tgaataaatt gcaatttcat ttgatgctcg atgagttttt
     1861 ctaagcggcg cgccatcgaa tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg
     1921 aagagagtca attcagggtg gtgaatatga aaccagtaac gttatacgat gtcgcagagt
     1981 atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg
     2041 cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg
     2101 tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg
     2161 ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg
     2221 ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc
     2281 acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg
     2341 atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg
     2401 accagacacc catcaacagt attattttct cccatgagga cggtacgcga ctgggcgtgg
     2461 agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg
     2521 tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc
     2581 cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa
     2641 tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg
     2701 gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg
     2761 gatacgacga taccgaagat agctcatgtt atatcccgcc gttaaccacc atcaaacagg
     2821 attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg
     2881 cggtgaaggg caatcagctg ttgccagtct cactggtgaa aagaaaaacc accctggcgc
     2941 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     3001 aggtttcccg actggaaagc gggcagtgac tcatgaccaa aatcccttaa cgtgagttac
     3061 gcgcgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3121 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3181 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3241 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttagcccacc acttcaagaa
     3301 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3361 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3421 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3481 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3541 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3601 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3661 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3721 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3781 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3841 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaaggcgaga gtagggaact
     3901 gccaggcatc aaactaagca gaaggcccct gacggatggc ctttttgcgt ttctacaaac
     3961 tctttctgtg ttgtaaaacg acggccagtc ttaagctcgg gccccctggg cggttctgat
     4021 aacgagtaat cgttaatccg caaataacgt aaaaacccgc ttcggcgggt ttttttatgg
     4081 ggggagttta gggaaagagc atttgtcaga atatttaagg gcgcctgtca ctttgcttga
     4141 tatatgagaa ttatttaacc ttataaatga gaaaaaagca acgcacttta aataagatac
     4201 gttgcttttt cgattgatga acacctataa ttaaactatt catctattat ttatgatttt
     4261 ttgtatatac aatatttcta gtttgttaaa gagaattaag aaaataaatc tcgaaaataa
     4321 taaagggaaa atcagttttt gatatcaaaa ttatacatgt caacgataat acaaaatata
     4381 atacaaacta taagatgtta tcagtattta ttatgcattt agaataaatt ttgtgtcgcc
     4441 cttccgcgaa at
//
