LOCUS       sequence-208176-        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: pHBS934 TDP43 RRM-GFP M-V
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..45
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     27..44
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    100..124
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      143..152
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1652..1676)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1721..1855
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1745..1764)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1799..1818
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1968..1988)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1968..1986)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2007..2023)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2007..2023)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2020..2042)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2031..2047
                     /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(2055..2061,2062..2079,2080..2085))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2100..2121
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2238..2255)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2409..2997)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2489..2508)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3168..3959,3960..4028))
                     /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     3791..3810
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4029..4133)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4159..4614)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4296..4317)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4508..4527
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4685..5669
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 tgcctgggga cgtcggagca agcttgattt aggtgacact atagaataca agctacttgt
       61 tctttttgca ggatcccatc gattcgaatt caaggatcaa caagtttgta caaaaaagca
      121 ggctggcgcc ggtaccaccg gtgccaccat gggatccatg tctgaatata ttcgggtaac
      181 cgaagatgag aacgatgagc ccattgaaat accatcggaa gacgatggga cggtgctgct
      241 ctccacggtt acagcccagt ttccaggggc gtgtgggctt cgctacagga atccagtgtc
      301 tcagtgtatg agaggtgtcc ggctggtaga aggaattctg catgccccag atgctggctg
      361 gggaaatctg gtgtatgttg tcaactatcc aaaagataac aaaagaaaaa tggatgagac
      421 agatgcttca tcagcagtga aagtgaaaag agcagtccag aaaacatcca gtaaaggaga
      481 ggaactattt acaggtactg tgcctatcaa agtcgagctg gacggcgacg taaacggcca
      541 caagttcagc gtgcgtggcg agggcgaggg cgatgccacc gagggcaagc tgaccctgaa
      601 gttcatctgc accaccggca agctgccggt gccgtggccg accctcgtga ccaccctgac
      661 ctacggcgtg cagtgcttca gccgctaccc ggaccacatg aagcgtcacg acttcttcaa
      721 gtccgccatg ccggaaggct acgtccagga gcgcaccatc gagttcaagg acgacggcac
      781 ctacaagacc cgcgcggagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
      841 gaagggcaac gacttcaagg aggacggcaa catcctgggt cacaagctgg agtacaacca
      901 caacagccac aacgtccgta tcgaagccga caagcagaag aacggcatca aggctaactt
      961 caagatccgc cacaacgttg aggacggcag ccagcaggaa gccgaccaca aacagcagaa
     1021 caccccgatc ggcgacggcc cggtgcgcct gccggacaac cactatctga gcacccagac
     1081 cacgctgagc aaagacccga acgagaagcg cgatcacatg gtcctgaagg agttcgtgac
     1141 ggcagccggt atcactaaag gtgaagatga aagtggatcc aatgccgaac ctaagcataa
     1201 tagcaatcgg cagctggagc gcagtggccg cttcggcgga aatcctggcg gttttggcaa
     1261 tcaaggggga tttggaaact cgaggggcgg aggggccggc ttaggcaaca accaaggatc
     1321 aaacgtaggg gggggcgtca actttggggc tttttctatt aatcccgcta tgatggcagc
     1381 agcgcaagca gcgttacagt cgtcgtgggg gatgatgggg atgttagctt cgcagcagaa
     1441 ccagtctggg ccttccggca acaatcagaa tcagggaaat gtccaacgtg agccgaacca
     1501 ggcatttggg tccggaaata acagctattc aggatctaat agcggcgctg caattggttg
     1561 gggctccgcc tcaaatgcgg gctcgggttc tggttttaac ggtggattcg ggtcaagcgt
     1621 ggacagtaag tcttcgggtt ggggcgttta gacccagctt tcttgtacaa agtggttgat
     1681 cctctcgagc ctctagaact atagtgagtc gtattacgta gatccagaca tgataagata
     1741 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     1801 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     1861 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaatt
     1921 cgcggccgcg gcgccaatgc attgggcccg gtacccagct tttgttccct ttagtgaggg
     1981 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     2041 ctcacaattc cacacaacat acgagccggg agcataaagt gtaaagcctg gggtgcctaa
     2101 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2161 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2221 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2281 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2341 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2401 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2461 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2521 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2581 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2641 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2701 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2761 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2821 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2881 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2941 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3001 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3061 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3121 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3181 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3241 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3301 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3361 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3421 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3481 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3541 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3601 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3661 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3721 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3781 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3841 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3901 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3961 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4021 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4081 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4141 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4201 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     4261 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4321 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     4381 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4441 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4501 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4561 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
     4621 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     4681 gccagtcgac cgccaattca atatggcgta tatggactca tgccaattca atatggtgga
     4741 tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga
     4801 tctggacccc agccaattca atatggcgga cttggcacca tgccaattca atatggcgga
     4861 cctggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg
     4921 tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat
     4981 attggctata tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg
     5041 gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg
     5101 gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg
     5161 cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct
     5221 ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt
     5281 caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc
     5341 attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac
     5401 gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc
     5461 aagtacatta ctattggaag tacgccaggg tacattggca gtactcccat tgacgtcaat
     5521 ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg
     5581 caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg
     5641 tctatataag caatgctcgt ttagggaacc gccattc
//
