LOCUS       sequence-207994-        5677 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To test the effect of sequence on TDP43 droplet properties.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5677)
  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 5677)
  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: pHBS1052 TDP43 RRM-GFP R-K
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        42..60
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     42..59
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    115..139
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      158..167
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1667..1691)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1736..1870
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1760..1779)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1814..1833
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1983..2003)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1983..2001)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2022..2038)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2022..2038)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2035..2057)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2046..2062
                     /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)."
     promoter        complement(join(2070..2076,2077..2094,2095..2100))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2115..2136
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2253..2270)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2424..3012)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2504..2523)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3183..3974,3975..4043))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3806..3825
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4044..4148)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4174..4629)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4311..4332)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4523..4542
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(4700..5677,1..7)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 agggaaccgc cattctgcct ggggacgtcg gagcaagctt gatttaggtg acactataga
       61 atacaagcta cttgttcttt ttgcaggatc ccatcgattc gaattcaagg atcaacaagt
      121 ttgtacaaaa aagcaggctg gcgccggtac caccggtgcc accatgggat ccatgtctga
      181 atatattcgg gtaaccgaag atgagaacga tgagcccatt gaaataccat cggaagacga
      241 tgggacggtg ctgctctcca cggttacagc ccagtttcca ggggcgtgtg ggcttcgcta
      301 caggaatcca gtgtctcagt gtatgagagg tgtccggctg gtagaaggaa ttctgcatgc
      361 cccagatgct ggctggggaa atctggtgta tgttgtcaac tatccaaaag ataacaaaag
      421 aaaaatggat gagacagatg cttcatcagc agtgaaagtg aaaagagcag tccagaaaac
      481 atccagtaaa ggagaggaac tatttacagg tactgtgcct atcaaagtcg agctggacgg
      541 cgacgtaaac ggccacaagt tcagcgtgcg tggcgagggc gagggcgatg ccaccgaggg
      601 caagctgacc ctgaagttca tctgcaccac cggcaagctg ccggtgccgt ggccgaccct
      661 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc tacccggacc acatgaagcg
      721 tcacgacttc ttcaagtccg ccatgccgga aggctacgtc caggagcgca ccatcgagtt
      781 caaggacgac ggcacctaca agacccgcgc ggaggtgaag ttcgagggcg acaccctggt
      841 gaaccgcatc gagctgaagg gcaacgactt caaggaggac ggcaacatcc tgggtcacaa
      901 gctggagtac aaccacaaca gccacaacgt ccgtatcgaa gccgacaagc agaagaacgg
      961 catcaaggct aacttcaaga tccgccacaa cgttgaggac ggcagccagc aggaagccga
     1021 ccacaaacag cagaacaccc cgatcggcga cggcccggtg cgcctgccgg acaaccacta
     1081 tctgagcacc cagaccacgc tgagcaaaga cccgaacgag aagcgcgatc acatggtcct
     1141 gaaggagttc gtgacggcag ccggtatcac taaaggtgaa gatgaaagtg gatccaatgc
     1201 cgaacctaag cacaattcaa acaagcaact agagaaaagt gggaagttcg gcggcaaccc
     1261 tggtggattc ggtaaccaag gcgggttcgg caacagtaag gggggcggcg ccggacttgg
     1321 aaacaaccag ggttcaaata tgggcggcgg catgaatttt ggagctttca gcatcaatcc
     1381 agctatgatg gcggctgccc aggctgctct gcaatcttct tgggggatga tgggtatgct
     1441 tgcaagtcaa cagaatcagt ccggtccatc cggaaataac cagaaccagg gtaatatgca
     1501 gaaagagcct aatcaagcgt ttggcagtgg taacaattcg tattctgggt ccaatagcgg
     1561 agctgctata ggttggggct cagcaagcaa tgccggtagt ggtagcggct ttaatggagg
     1621 ctttggtagt tctatggact caaagtcttc cgggtggggc atgtagaccc agctttcttg
     1681 tacaaagtgg ttgatcctct cgagcctcta gaactatagt gagtcgtatt acgtagatcc
     1741 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     1801 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     1861 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     1921 ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt
     1981 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     2041 tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa
     2101 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     2161 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     2221 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     2281 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     2341 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2401 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2461 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2521 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2581 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2641 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2701 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2761 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2821 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2881 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2941 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3001 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3061 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3121 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3181 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3241 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3301 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3361 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3421 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3481 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3541 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3601 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3661 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3721 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3781 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3841 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3901 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3961 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4021 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4081 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4141 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     4201 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     4261 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     4321 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     4381 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     4441 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     4501 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     4561 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     4621 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     4681 cctcttcgct attacgccag tcgaccgcca attcaatatg gcgtatatgg actcatgcca
     4741 attcaatatg gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca
     4801 attcaatatg gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca
     4861 attcaatatg gcggacctgg cactgtgcca actggggagg ggtctacttg gcacggtgcc
     4921 aagtttgagg aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc
     4981 aataatggcg gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg
     5041 ccctatgcca atatggctat tggccaggtt caatactatg tattggccct atgccatata
     5101 gtattccata tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat
     5161 ataccatata tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt
     5221 ctctatatat ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc
     5281 ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca
     5341 ataggaccac ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc
     5401 acgcccccta ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat
     5461 taatagtaac ttggcaagta cattactatt ggaagtacgc cagggtacat tggcagtact
     5521 cccattgacg tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac
     5581 gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc
     5641 gtgcctaatg ggaggtctat ataagcaatg ctcgttt
//
