LOCUS       sequence-229687-        5677 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To test the effect of sequence on TDP43 RRM-GFP 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: pHBS1385 TDP43 RRM-GFP VLIM-S
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        70..1054
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1089..1107
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1089..1106
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1162..1186
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1205..1214
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2714..2738)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2783..2917
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2807..2826)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2861..2880
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3030..3050)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3030..3048)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3069..3085)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3069..3085)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3082..3104)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3093..3109
                     /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(3117..3123,3124..3141,3142..3147))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3162..3183
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3300..3317)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3471..4059)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3551..3570)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4230..5021,5022..5090))
                     /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     4853..4872
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5091..5195)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5221..5676)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5358..5379)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5570..5589
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag tcgaccgcca attcaatatg gcgtatatgg actcatgcca attcaatatg
      121 gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg
      181 gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg
      241 gcggacctgg cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg
      301 aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg
      361 gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca
      421 atatggctat tggccaggtt caatactatg tattggccct atgccatata gtattccata
      481 tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata
      541 tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt ctctatatat
      601 ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt
      661 gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac
      721 ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta
      781 ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac
      841 ttggcaagta cattactatt ggaagtacgc cagggtacat tggcagtact cccattgacg
      901 tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg
      961 aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg
     1021 ggaggtctat ataagcaatg ctcgtttagg gaaccgccat tctgcctggg gacgtcggag
     1081 caagcttgat ttaggtgaca ctatagaata caagctactt gttctttttg caggatccca
     1141 tcgattcgaa ttcaaggatc aacaagtttg tacaaaaaag caggctggcg ccggtaccac
     1201 cggtgccacc atgggatcca tgtctgaata tattcgggta accgaagatg agaacgatga
     1261 gcccattgaa ataccatcgg aagacgatgg gacggtgctg ctctccacgg ttacagccca
     1321 gtttccaggg gcgtgtgggc ttcgctacag gaatccagtg tctcagtgta tgagaggtgt
     1381 ccggctggta gaaggaattc tgcatgcccc agatgctggc tggggaaatc tggtgtatgt
     1441 tgtcaactat ccaaaagata acaaaagaaa aatggatgag acagatgctt catcagcagt
     1501 gaaagtgaaa agagcagtcc agaaaacatc cagtaaagga gaggaactat ttacaggtac
     1561 tgtgcctatc aaagtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgcgtgg
     1621 cgagggcgag ggcgatgcca ccgagggcaa gctgaccctg aagttcatct gcaccaccgg
     1681 caagctgccg gtgccgtggc cgaccctcgt gaccaccctg acctacggcg tgcagtgctt
     1741 cagccgctac ccggaccaca tgaagcgtca cgacttcttc aagtccgcca tgccggaagg
     1801 ctacgtccag gagcgcacca tcgagttcaa ggacgacggc acctacaaga cccgcgcgga
     1861 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca acgacttcaa
     1921 ggaggacggc aacatcctgg gtcacaagct ggagtacaac cacaacagcc acaacgtccg
     1981 tatcgaagcc gacaagcaga agaacggcat caaggctaac ttcaagatcc gccacaacgt
     2041 tgaggacggc agccagcagg aagccgacca caaacagcag aacaccccga tcggcgacgg
     2101 cccggtgcgc ctgccggaca accactatct gagcacccag accacgctga gcaaagaccc
     2161 gaacgagaag cgcgatcaca tggtcctgaa ggagttcgtg acggcagccg gtatcactaa
     2221 aggtgaagat gaaagtggat ccaatgccga acctaagcac aacagtaatc gacagtccga
     2281 gagaagcggg cggttcggtg gtaatccagg ggggttcgga aaccaaggag gttttggtaa
     2341 ctctagggga ggaggcgctg gttccggtaa taaccaaggc agcaacagtg gggggggcag
     2401 taatttcggt gctttttcaa gtaacccagc aatgatggcc gccgcacaag cagctttgca
     2461 aagttcttgg ggcatgatgg gcatgctggc gtcacagcaa aatcaaagcg gaccctcagg
     2521 aaataatcaa aaccaaggca atagccagag ggagcctaat caggccttcg gatcaggcaa
     2581 taattcatac agtggttcaa actccggggc agcctctggg tggggcagcg caagtaatgc
     2641 cggaagcggt agtggattca acggagggtt cgggtcaagt agcgatagta aaagttccgg
     2701 ttggggctct tagacccagc tttcttgtac aaagtggttg atcctctcga gcctctagaa
     2761 ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg agtttggaca
     2821 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2881 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2941 tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcgcggccg cggcgccaat
     3001 gcattgggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     3061 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3121 atacgagccg ggagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3181 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3241 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3301 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3361 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3421 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3481 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3541 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3601 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3661 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3721 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3781 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccaccgg taacaggatt
     3841 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3901 tacactagaa gaacagtatt tggtatctgc gctttgctga agccagttac cttcggaaaa
     3961 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4021 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4081 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4141 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4201 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4261 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4321 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4381 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4441 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4501 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4561 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4621 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4681 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4741 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4801 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4861 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4921 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4981 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5041 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5101 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5161 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5221 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     5281 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     5341 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     5401 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     5461 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     5521 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     5581 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     5641 cacccgccgc gcttaatgcg ccgctacagg gcgcgtc
//
