LOCUS       sequence-261741-        6235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of TDP43 with WT CTD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6235)
  AUTHORS   Schmidt HB, Barreau A, Rohatgi R
  TITLE     Phase separation-deficient TDP43 remains functional in splicing.
  JOURNAL   Nat Commun. 2019 Oct 25;10(1):4890. doi: 10.1038/s41467-019-12740-2.
  PUBMED    31653829
REFERENCE   2  (bases 1 to 6235)
  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: "Nat 
            Commun."; date: "2019-10-25"; volume: "10(1):4890. doi"; pages: " 
            10.1038/s41467-019-12740-2"
COMMENT     Sequence Label: pHBS834 H14-SUMO-TDP43 WT-TEV-mCherry
FEATURES             Location/Qualifiers
     source          1..6235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    8..24
                     /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)."
     RBS             43..54
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1678..1698
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1702..2406
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="SKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQ
                     TAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFED
                     GGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGE
                     IKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEG
                     RHSTGGMDELYK"
     primer_bind     complement(1841..1859)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2109..2128
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      2426..2520
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2568..2587)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2579..3394
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2579..3394
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2652..2671)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3613..3699
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3792..4874)
                     /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     4836..4855
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4875..4952)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(5036..5058)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5144..5284
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5199..5218
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5299..5316)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5470..6058)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5550..5569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6149..6167
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(6191..6205,6206..6211,6212..6228,6229..6235)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    6211..6227
                     /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)."
ORIGIN
        1 gattcaattg tgagcggata acaatttcac acagaattca ttaaagagga gaaattaacc
       61 atgagcaagc atcaccatca ttcaggccat caccataccg gacaccacca tcattcaggc
      121 agtcatcacc attccggatc tgctgcgggt ggcgaagaag ataagaaacc ggcaggtggc
      181 gaaggtggcg gtgcccatat caacctgaaa gtgaaaggtc aagacggcaa cgaagtcttt
      241 ttccgcatca aacgttctac ccagctgaaa aagctgatga acgcatactg tgaccgtcag
      301 tctgtagaca tgaccgcaat tgctttcctg tttgatggtc gtcgcctgcg tgcggaacag
      361 accccggatg aactggagat ggaagatggc gacgaaatcg acgcaatgct gcaccagact
      421 ggtggcggat ccatgtctga atatattcgg gtaaccgaag atgagaacga tgagcccatt
      481 gaaataccat cggaagacga tgggacggtg ctgctctcca cggttacagc ccagtttcca
      541 ggggcgtgtg ggcttcgcta caggaatcca gtgtctcagt gtatgagagg tgtccggctg
      601 gtagaaggaa ttctgcatgc cccagatgct ggctggggaa atctggtgta tgttgtcaac
      661 tatccaaaag ataacaaaag aaaaatggat gagacagatg cttcatcagc agtgaaagtg
      721 aaaagagcag tccagaaaac atccgattta atagtgttgg gtctcccatg gaaaacaacc
      781 gaacaggacc tgaaagagta ttttagtacc tttggagaag ttcttatggt gcaggtcaag
      841 aaagatctta agactggtca ttcaaagggg tttggctttg ttcgttttac ggaatatgaa
      901 acacaagtga aagtaatgtc acagcgacat atgatagatg gacgatggtg tgactgcaaa
      961 cttcctaatt ctaagcaaag ccaagatgag cctttgagaa gcagaaaagt gtttgtgggg
     1021 cgctgtacag aggacatgac tgaggatgag ctgcgggagt tcttctctca gtacggggat
     1081 gtgatggatg tcttcatccc caagccattc agggcctttg cctttgttac atttgcagat
     1141 gatcagattg cgcagtctct ttgtggagag gacttgatca ttaaaggaat cagcgttcat
     1201 atatccaatg ccgaacctaa gcacaatagc aatagacagt tagaaagaag tggaagattt
     1261 ggtggtaatc caggtggctt tgggaatcag ggtggatttg gtaatagcag agggggtgga
     1321 gctggtttgg gaaacaatca aggtagtaat atgggtggtg ggatgaactt tggtgcgttc
     1381 agcattaatc cagccatgat ggctgccgcc caggcagcac tacagagcag ttggggtatg
     1441 atgggcatgt tagccagcca gcagaaccag tcaggcccat cgggtaataa ccaaaaccaa
     1501 ggcaacatgc agagggagcc aaaccaggcc ttcggttctg gaaataactc ttatagtggc
     1561 tctaattctg gtgcagcaat tggttgggga tcagcatcca atgcagggtc gggcagtggt
     1621 tttaatggag gctttggctc aagcatggat tctaagtctt ctggctgggg aatgggcgag
     1681 aacctgtatt ttcagggtac cagcaagggc gaggaggata acatggccat catcaaggag
     1741 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag
     1801 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     1861 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
     1921 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc
     1981 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
     2041 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc
     2101 tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg
     2161 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc
     2221 ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     2281 ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc
     2341 gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac
     2401 aagtaagctt aattagctga gcttggactc ctgttgatag atccagtaat gacctcagaa
     2461 ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat tggtgagaat
     2521 ccaagctagc catgaaaata aactgtctgc ttacataaac agtaatacaa ggggtgttat
     2581 gagccatatt caacgggaaa cgtcttgctc taggccgcga ttaaattcca acatggatgc
     2641 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     2701 tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     2761 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     2821 tccgaccatc aagcatttta tccgtactcc tgatgatgct tggttactca cgactgcgat
     2881 ccccggcaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     2941 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     3001 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     3061 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     3121 aatgcacaaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     3181 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     3241 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     3301 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     3361 gcagtttcat ttgatgctcg atgagttttt ctaagaatta attcatgggc aaatattata
     3421 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
     3481 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
     3541 gggcgtaatt tttttaaggc agttattggt gcccttaaac gcctggggta atgactctct
     3601 agcttgaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     3661 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccctctag attacgtgca
     3721 gtcgatgata agctgtcaaa catgagaatt gtgcctaatg agtgagctaa cttacattaa
     3781 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     3841 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gggccagggt ggtttttctt
     3901 ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc
     3961 agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac
     4021 ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca
     4081 ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg
     4141 gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa
     4201 ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg
     4261 agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct
     4321 aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg
     4381 tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac
     4441 gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag
     4501 ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct
     4561 tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga
     4621 gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg
     4681 ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc
     4741 agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg
     4801 ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac
     4861 gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata
     4921 ccgcgaaagg ttttgcacca ttcgatggtg tcggaatttc gggcagcgtt gggtcctggc
     4981 cacgggtgcg catgatctag agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     5041 gacacatgca gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagcagac
     5101 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     5161 cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact
     5221 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     5281 caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5341 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5401 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5461 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5521 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5581 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5641 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5701 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5761 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5821 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5881 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     5941 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6001 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6061 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6121 gattttggtc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     6181 cctcgagaaa tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taata
//
