LOCUS       sequence-201398-        5283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of TDP-43 NTD Y4R.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5283)
  AUTHORS   Wang A, Conicella AE, Schmidt HB, Martin EW, Rhoads SN, Reeb AN, 
            Nourse A, Ramirez Montero D, Ryan VH, Rohatgi R, Shewmaker F, Naik 
            MT, Mittag T, Ayala YM, Fawzi NL
  TITLE     A single N-terminal phosphomimic disrupts TDP-43 polymerization, 
            phase separation, and RNA splicing.
  JOURNAL   EMBO J. 2018 Mar 1;37(5). pii: embj.201797452. doi: 
            10.15252/embj.201797452. Epub 2018 Feb 9.
  PUBMED    29438978
REFERENCE   2  (bases 1 to 5283)
  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: "EMBO J."; 
            date: "2018-03-1"; volume: "37(5). pii: embj.201797452. doi"; pages:
            " 10.15252/embj.201797452. Epub 2018 Feb 9"
COMMENT     Sequence Label: TDP-43_NTD1-80_Y4R
FEATURES             Location/Qualifiers
     source          1..5283
                     /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      1438..1485
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1708..1727)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        1794..1885
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1886..2695
                     /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             2778..3860
                     /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(2797..2816)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      3946..4534
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4435..4454
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4688..4705
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4803..4820
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4804..4820
                     /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 tgagcgagag gattcgtgtt accgaagatg
      181 agaacgacga gccgatcgag attccgagcg aagatgacgg tacggtcttg ctgagcacgg
      241 tgaccgcgca gttcccaggc gcctgtggcc tgcgctatcg taatccggtc agccagtgca
      301 tgcgcggtgt ccgtctggtt gagggcattc tgcacgctcc ggacgcgggc tggggcaatc
      361 tggtgtacgt ggtcaattac ccgaaagact agaagcgcaa gatggatgaa actgacgcca
      421 gctctgcggt taaagtcaag cgtgcagtgt agaaaacgag cgacctggtc aagcgtgcag
      481 tgtagaaaac gagcgacctg atcgtgctgg gtttgccgtg gaaaaccacg gagcaagacc
      541 tgaaagaata ctttagcacc ttcggtgagg tgctgatggt ccaggtaaag aaagatctga
      601 aaaccggtca cagcaagggt tttggcttcg tccgtttcac cgagtatgaa acccaggtga
      661 aagttatgtc gcaacgtcac atgatcgacg gtcgttggtg tgactgcaag ctgccgaaca
      721 gcaagcagag ccaagatgaa ccgctgcgct cccgtaaagt tttcgttggt cgctgcaccg
      781 aagatatgac ggaagatgag ctgcgtgagt tcttttctca atatggtgac gtgatggatg
      841 ttttcatccc gaaacctttc cgcgcctttg catttgttac cttcgctgac gatcaaatcg
      901 cgcagagcct gtgtggtgag gatctgatta tcaaaggtat ttccgttcac atttctaacg
      961 cggagccaaa gcataacagc aaccgccaac tggaacgttc cggtcgtttc ggtggtaatc
     1021 cgggtggttt cggcaaccag ggtggctttg gcaattcgcg tggcggtggt gccggcttgg
     1081 gcaataacca aggttccaat atgggtggcg gtatgaactt tggtgccttc tccatcaatc
     1141 cggcaatgat ggcggctgca caagcggcgc tgcagtctag ctggggtatg atgggcatgc
     1201 tggcgagcca gcagaaccag tcaggcccga gcggcaataa tcagaatcaa ggtaacatgc
     1261 agcgtgaacc gaatcaagcg tttggtagcg gtaacaatag ctacagcggt agcaacagcg
     1321 gtgcggcaat cggctggggt agcgcaagca atgcgggcag cggtagcggt ttcaacggcg
     1381 gctttggtag cagcatggat agcaagagct ccggttgggg catgtaactc gagcccccta
     1441 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgcccct gagacgcgtc
     1501 aatcgagttc gtacctaagg gcgacacccc ctaattagcc cgggcgaaag gcccagtctt
     1561 tcgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc atggggagtc
     1621 cccacactac catcggcgct acggcgtttc acttctgagt tcggcatggg gtcaggtggg
     1681 accaccgcgc tactgccgcc aggcaaacaa ggggtgttat gagccatatt caggtataaa
     1741 tgggctcgcg ataatgttca gaattggtta attggttgta acactgaccc ctatttgttt
     1801 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1861 tcaataatat tgaaaaagga agaatatgag ccatattcaa cgggaaacgt cgaggccgcg
     1921 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     1981 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     2041 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     2101 gctgacggaa tttatgccac ttccgaccat caagcatttt atccgtactc ctgatgatgc
     2161 atggttactc accactgcga tccccggaaa aacagcgttc caggtattag aagaatatcc
     2221 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcactcgat
     2281 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgcctcgctc aggcgcaatc
     2341 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     2401 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     2461 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     2521 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     2581 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     2641 taatcctgat atgaataaat tgcaatttca tttgatgctc gatgagtttt tctaagcggc
     2701 gcgccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
     2761 aattcagggt ggtgaatatg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
     2821 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
     2881 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
     2941 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
     3001 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
     3061 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
     3121 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
     3181 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
     3241 ccatcaacag tattattttc tcccatgagg acggtacgcg actgggcgtg gagcatctgg
     3301 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc
     3361 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg
     3421 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg
     3481 agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc
     3541 gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg
     3601 ataccgaaga tagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc
     3661 tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg
     3721 gcaatcagct gttgccagtc tcactggtga aaagaaaaac caccctggcg cccaatacgc
     3781 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     3841 gactggaaag cgggcagtga ctcatgacca aaatccctta acgtgagtta cgcgcgcgtc
     3901 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3961 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4021 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4081 accaaatact gttcttctag tgtagccgta gttagcccac cacttcaaga actctgtagc
     4141 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4201 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4261 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4321 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4381 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4441 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4501 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4561 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4621 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4681 cgagcgcagc gagtcagtga gcgaggaagc ggaaggcgag agtagggaac tgccaggcat
     4741 caaactaagc agaaggcccc tgacggatgg cctttttgcg tttctacaaa ctctttctgt
     4801 gttgtaaaac gacggccagt cttaagctcg ggccccctgg gcggttctga taacgagtaa
     4861 tcgttaatcc gcaaataacg taaaaacccg cttcggcggg tttttttatg gggggagttt
     4921 agggaaagag catttgtcag aatatttaag ggcgcctgtc actttgcttg atatatgaga
     4981 attatttaac cttataaatg agaaaaaagc aacgcacttt aaataagata cgttgctttt
     5041 tcgattgatg aacacctata attaaactat tcatctatta tttatgattt tttgtatata
     5101 caatatttct agtttgttaa agagaattaa gaaaataaat ctcgaaaata ataaagggaa
     5161 aatcagtttt tgatatcaaa attatacatg tcaacgataa tacaaaatat aatacaaact
     5221 ataagatgtt atcagtattt attatgcatt tagaataaat tttgtgtcgc ccttccgcga
     5281 aat
//
