LOCUS       sequence-256252-        6236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of TDP43 with F-S mutant CTD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6236)
  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 6236)
  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: pHBS1551 H14-SUMO-TDP43 F-S-TEV-mCherry
FEATURES             Location/Qualifiers
     source          1..6236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..24,25..30,31..47,48..54)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /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    62..78
                     /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             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1732..1752
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1756..2460
                     /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(1895..1913)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2163..2182
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      2480..2574
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2622..2641)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2633..3448
                     /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             2633..3448
                     /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(2706..2725)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3667..3753
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3847..4929)
                     /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     4891..4910
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4930..5007)
                     /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(5091..5113)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5199..5339
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5254..5273
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5354..5371)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5525..6113)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5605..5624)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6204..6222
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aaccatgagc
      121 aagcatcacc atcattcagg ccatcaccat accggacacc accatcattc aggcagtcat
      181 caccattccg gatctgctgc gggtggcgaa gaagataaga aaccggcagg tggcgaaggt
      241 ggcggtgccc atatcaacct gaaagtgaaa ggtcaagacg gcaacgaagt ctttttccgc
      301 atcaaacgtt ctacccagct gaaaaagctg atgaacgcat actgtgaccg tcagtctgta
      361 gacatgaccg caattgcttt cctgtttgat ggtcgtcgcc tgcgtgcgga acagaccccg
      421 gatgaactgg agatggaaga tggcgacgaa atcgacgcaa tgctgcacca gactggtggc
      481 ggatccatgt ctgaatatat tcgggtaacc gaagatgaga acgatgagcc cattgaaata
      541 ccatcggaag acgatgggac ggtgctgctc tccacggtta cagcccagtt tccaggggcg
      601 tgtgggcttc gctacaggaa tccagtgtct cagtgtatga gaggtgtccg gctggtagaa
      661 ggaattctgc atgccccaga tgctggctgg ggaaatctgg tgtatgttgt caactatcca
      721 aaagataaca aaagaaaaat ggatgagaca gatgcttcat cagcagtgaa agtgaaaaga
      781 gcagtccaga aaacatccga tttaatagtg ttgggtctcc catggaaaac aaccgaacag
      841 gacctgaaag agtattttag tacctttgga gaagttctta tggtgcaggt caagaaagat
      901 cttaagactg gtcattcaaa ggggtttggc tttgttcgtt ttacggaata tgaaacacaa
      961 gtgaaagtaa tgtcacagcg acatatgata gatggacgat ggtgtgactg caaacttcct
     1021 aattctaagc aaagccaaga tgagcctttg agaagcagaa aagtgtttgt ggggcgctgt
     1081 acagaggaca tgactgagga tgagctgcgg gagttcttct ctcagtacgg ggatgtgatg
     1141 gatgtcttca tccccaagcc attcagggcc tttgcctttg ttacatttgc agatgatcag
     1201 attgcgcagt ctctttgtgg agaggacttg atcattaaag gaatcagcgt tcatatatcc
     1261 aatgccgaac ctaaacataa ttcaaaccgg cagttggaaa gatcggggcg cagtggtggc
     1321 aacccaggcg gttctggtaa tcagggcggc tcgggaaatt ccagaggcgg gggtgcaggg
     1381 cttggcaaca atcaggggtc caatatgggc ggtggtatga actcaggtgc cagctcgatt
     1441 aatccggcca tgatggcggc agcgcaggcg gcgctgcagt cttcctgggg tatgatgggt
     1501 atgttagcat cccaacagaa ccagagtggc ccgtcgggga ataaccaaaa tcaggggaat
     1561 atgcagcgcg aaccgaatca agcttcaggt agtggtaaca acagttatag tggatcgaac
     1621 agtggcgccg caattggatg gggttccgca tccaatgcag gctcaggcag cggctcgaat
     1681 ggcggctcag gttctagtat ggattcgaag agctctggtt ggggaatggg cgagaacctg
     1741 tattttcagg gtaccagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
     1801 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     1861 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
     1921 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     1981 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     2041 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     2101 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     2161 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc
     2221 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     2281 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     2341 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
     2401 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa
     2461 gcttaattag ctgagcttgg actcctgttg atagatccag taatgacctc agaactccat
     2521 ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc
     2581 tagccatgaa aataaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     2641 tattcaacgg gaaacgtctt gctctaggcc gcgattaaat tccaacatgg atgctgattt
     2701 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt
     2761 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     2821 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     2881 catcaagcat tttatccgta ctcctgatga tgcttggtta ctcacgactg cgatccccgg
     2941 caaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     3001 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     3061 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     3121 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     3181 caaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     3241 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     3301 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     3361 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     3421 tcatttgatg ctcgatgagt ttttctaaga attaattcat gggcaaatat tatacgcaag
     3481 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     3541 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     3601 aattttttta aggcagttat tggtgccctt aaacgcctgg ggtaatgact ctctagcttg
     3661 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     3721 ttgtcggtga acgctctcct gagtaggaca aatccgccgc tctagattac gtgcagtcga
     3781 tgataagctg tcaaacatga gaattgtgcc taatgagtga gctaacttac attaattgcg
     3841 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3901 ggccaacgcg cggggagagg cggtttgcgt attgggggcc agggtggttt ttcttttcac
     3961 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
     4021 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
     4081 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac
     4141 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
     4201 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
     4261 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
     4321 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
     4381 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
     4441 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
     4501 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
     4561 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
     4621 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
     4681 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
     4741 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
     4801 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
     4861 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
     4921 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
     4981 aaaggttttg caccattcga tggtgtcgga atttcgggca gcgttgggtc ctggccacgg
     5041 gtgcgcatga tctagagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     5101 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     5161 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     5221 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
     5281 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     5341 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5401 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5461 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5521 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5581 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5641 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5701 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5761 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5821 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5881 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5941 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6001 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6061 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6121 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6181 tggtcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcac
//
