LOCUS       sequence-261740-        6235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of TDP43 with FYW-S mutant 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: pHBS1552 H14-SUMO-TDP43 FYW-S-TEV-mCherry
FEATURES             Location/Qualifiers
     source          1..6235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..15,16..21,22..38,39..45)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    21..37
                     /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)."
     protein_bind    53..69
                     /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             88..99
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1723..1743
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1747..2451
                     /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(1886..1904)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2154..2173
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      2471..2565
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2613..2632)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2624..3439
                     /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             2624..3439
                     /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(2697..2716)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3658..3744
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3837..4919)
                     /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     4881..4900
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4920..4997)
                     /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(5081..5103)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5189..5329
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5244..5263
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5344..5361)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5515..6103)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5595..5614)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6194..6212
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taatagattc aattgtgagc
       61 ggataacaat ttcacacaga attcattaaa gaggagaaat taaccatgag caagcatcac
      121 catcattcag gccatcacca taccggacac caccatcatt caggcagtca tcaccattcc
      181 ggatctgctg cgggtggcga agaagataag aaaccggcag gtggcgaagg tggcggtgcc
      241 catatcaacc tgaaagtgaa aggtcaagac ggcaacgaag tctttttccg catcaaacgt
      301 tctacccagc tgaaaaagct gatgaacgca tactgtgacc gtcagtctgt agacatgacc
      361 gcaattgctt tcctgtttga tggtcgtcgc ctgcgtgcgg aacagacccc ggatgaactg
      421 gagatggaag atggcgacga aatcgacgca atgctgcacc agactggtgg cggatccatg
      481 tctgaatata ttcgggtaac cgaagatgag aacgatgagc ccattgaaat accatcggaa
      541 gacgatggga cggtgctgct ctccacggtt acagcccagt ttccaggggc gtgtgggctt
      601 cgctacagga atccagtgtc tcagtgtatg agaggtgtcc ggctggtaga aggaattctg
      661 catgccccag atgctggctg gggaaatctg gtgtatgttg tcaactatcc aaaagataac
      721 aaaagaaaaa tggatgagac agatgcttca tcagcagtga aagtgaaaag agcagtccag
      781 aaaacatccg atttaatagt gttgggtctc ccatggaaaa caaccgaaca ggacctgaaa
      841 gagtatttta gtacctttgg agaagttctt atggtgcagg tcaagaaaga tcttaagact
      901 ggtcattcaa aggggtttgg ctttgttcgt tttacggaat atgaaacaca agtgaaagta
      961 atgtcacagc gacatatgat agatggacga tggtgtgact gcaaacttcc taattctaag
     1021 caaagccaag atgagccttt gagaagcaga aaagtgtttg tggggcgctg tacagaggac
     1081 atgactgagg atgagctgcg ggagttcttc tctcagtacg gggatgtgat ggatgtcttc
     1141 atccccaagc cattcagggc ctttgccttt gttacatttg cagatgatca gattgcgcag
     1201 tctctttgtg gagaggactt gatcattaaa ggaatcagcg ttcatatatc caatgccgaa
     1261 cctaagcata atagcaacag acaactcgaa cgaagcgggc ggagtggggg caaccccggc
     1321 ggatctggta atcaaggcgg aagcggcaat agccggggtg ggggagccgg tctcggcaat
     1381 aatcaaggat ctaacatggg cggcggtatg aactctggcg catccagtat caacccggcc
     1441 atgatggcag cagcacaggc cgccctgcaa agttcatggg gaatgatggg tatgttggca
     1501 agtcagcaga accagtcagg accttccggg aataatcaaa accaagggaa catgcaacgc
     1561 gaacccaatc aagcctcagg atcaggcaat aatagcagtt ccggctcaaa ttccggcgcg
     1621 gcgatagggt ctggctccgc ctcaaatgcg ggaagcggat cagggagtaa cggaggatca
     1681 ggctcatcta tggattcaaa atccagcggc agtggtatgg gcgagaacct gtattttcag
     1741 ggtaccagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
     1801 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
     1861 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
     1921 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
     1981 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
     2041 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
     2101 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
     2161 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
     2221 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
     2281 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
     2341 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
     2401 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta agcttaatta
     2461 gctgagcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
     2521 ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag ctagccatga
     2581 aaataaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     2641 ggaaacgtct tgctctaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     2701 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa
     2761 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     2821 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     2881 ttttatccgt actcctgatg atgcttggtt actcacgact gcgatccccg gcaaaacagc
     2941 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     3001 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     3061 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     3121 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc acaaactttt
     3181 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     3241 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     3301 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     3361 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     3421 gctcgatgag tttttctaag aattaattca tgggcaaata ttatacgcaa ggcgacaagg
     3481 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     3541 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taattttttt
     3601 aaggcagtta ttggtgccct taaacgcctg gggtaatgac tctctagctt gaggcatcaa
     3661 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     3721 aacgctctcc tgagtaggac aaatccgccc tctagattac gtgcagtcga tgataagctg
     3781 tcaaacatga gaattgtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac
     3841 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     3901 cggggagagg cggtttgcgt attgggggcc agggtggttt ttcttttcac cagtgagacg
     3961 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
     4021 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
     4081 gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg
     4141 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
     4201 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
     4261 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
     4321 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
     4381 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
     4441 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
     4501 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
     4561 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
     4621 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
     4681 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
     4741 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
     4801 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
     4861 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
     4921 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
     4981 caccattcga tggtgtcgga atttcgggca gcgttgggtc ctggccacgg gtgcgcatga
     5041 tctagagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     5101 cggcatccgc ttacagacaa gctgtgaccg tctccgggag cagacaagcc cgtcagggcg
     5161 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     5221 gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     5281 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc
     5341 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5401 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5461 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5521 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5581 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5641 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5701 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5761 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5821 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5881 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5941 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6001 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6061 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6121 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgac
     6181 attaacctat aaaaataggc gtatcacgag gccctttcgt cttcacctcg agaaa
//
