LOCUS       sequence-207916-        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: pHBS1037 TDP43 RRM-GFP 6x-Phi
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..43
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     25..42
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    98..122
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      141..150
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1650..1674)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1719..1853
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1743..1762)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1797..1816
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1966..1986)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1966..1984)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2005..2021)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2005..2021)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2018..2040)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2029..2045
                     /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(2053..2059,2060..2077,2078..2083))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2098..2119
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2236..2253)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2407..2995)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2487..2506)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3166..3957,3958..4026))
                     /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     3789..3808
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4027..4131)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4157..4612)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4294..4315)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4506..4525
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4683..5667
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag ctacttgttc
       61 tttttgcagg atcccatcga ttcgaattca aggatcaaca agtttgtaca aaaaagcagg
      121 ctggcgccgg taccaccggt gccaccatgg gatccatgtc tgaatatatt cgggtaaccg
      181 aagatgagaa cgatgagccc attgaaatac catcggaaga cgatgggacg gtgctgctct
      241 ccacggttac agcccagttt ccaggggcgt gtgggcttcg ctacaggaat ccagtgtctc
      301 agtgtatgag aggtgtccgg ctggtagaag gaattctgca tgccccagat gctggctggg
      361 gaaatctggt gtatgttgtc aactatccaa aagataacaa aagaaaaatg gatgagacag
      421 atgcttcatc agcagtgaaa gtgaaaagag cagtccagaa aacatccagt aaaggagagg
      481 aactatttac aggtactgtg cctatcaaag tcgagctgga cggcgacgta aacggccaca
      541 agttcagcgt gcgtggcgag ggcgagggcg atgccaccga gggcaagctg accctgaagt
      601 tcatctgcac caccggcaag ctgccggtgc cgtggccgac cctcgtgacc accctgacct
      661 acggcgtgca gtgcttcagc cgctacccgg accacatgaa gcgtcacgac ttcttcaagt
      721 ccgccatgcc ggaaggctac gtccaggagc gcaccatcga gttcaaggac gacggcacct
      781 acaagacccg cgcggaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
      841 agggcaacga cttcaaggag gacggcaaca tcctgggtca caagctggag tacaaccaca
      901 acagccacaa cgtccgtatc gaagccgaca agcagaagaa cggcatcaag gctaacttca
      961 agatccgcca caacgttgag gacggcagcc agcaggaagc cgaccacaaa cagcagaaca
     1021 ccccgatcgg cgacggcccg gtgcgcctgc cggacaacca ctatctgagc acccagacca
     1081 cgctgagcaa agacccgaac gagaagcgcg atcacatggt cctgaaggag ttcgtgacgg
     1141 cagccggtat cactaaaggt gaagatgaaa gtggatccaa tgccgaacct aagcataaca
     1201 gtaataggca gctagagcgc tccgggcgcg gcaatccgtt tggcggcggg ttcggaaacc
     1261 agaattcaag gggtggcggc gccggagggg gatttggact gggtaataat cagggttcta
     1321 atatgggtgg cggtatgaac gcgtttgggt tcagcatcaa ccctgctatg atggcggctg
     1381 cacaggcggc tctgcagtca tcatggggta tgatgggcat gctggcgtca cagcagaacc
     1441 agtccggccc gagcggaaac aatcagaatc aaggtaatat gcagcgtgag ccaaaccagg
     1501 cgtttggata ttccagtggg aataactctg gaagtaatag cggcgcagcc atcgggtggg
     1561 gctctgccag caatgcggga tcagggagta atggctttgg cggttttggt gggtggggga
     1621 tgtcatcgat ggatagtaag tcaagctaga cccagctttc ttgtacaaag tggttgatcc
     1681 tctcgagcct ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca
     1741 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     1801 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     1861 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg
     1921 cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt
     1981 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2041 cacaattcca cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg
     2101 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2161 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2221 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2281 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2341 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2401 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2461 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2521 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2581 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2641 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2701 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2761 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2821 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2881 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2941 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3001 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3061 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3121 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3181 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3241 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3301 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3361 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3421 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3481 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3541 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3601 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3661 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3721 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3781 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3841 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3901 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3961 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4021 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4081 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4141 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     4201 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     4261 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     4321 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     4381 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     4441 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     4501 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     4561 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     4621 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     4681 cagtcgaccg ccaattcaat atggcgtata tggactcatg ccaattcaat atggtggatc
     4741 tggacctgtg ccaattcaat atggcgtata tggactcgtg ccaattcaat atggtggatc
     4801 tggaccccag ccaattcaat atggcggact tggcaccatg ccaattcaat atggcggacc
     4861 tggcactgtg ccaactgggg aggggtctac ttggcacggt gccaagtttg aggaggggtc
     4921 ttggccctgt gccaagtccg ccatattgaa ttggcatggt gccaataatg gcggccatat
     4981 tggctatatg ccaggatcaa tatataggca atatccaata tggccctatg ccaatatggc
     5041 tattggccag gttcaatact atgtattggc cctatgccat atagtattcc atatatgggt
     5101 tttcctattg acgtagatag cccctcccaa tgggcggtcc catataccat atatggggct
     5161 tcctaatacc gcccatagcc actcccccat tgacgtcaat ggtctctata tatggtcttt
     5221 cctattgacg tcatatgggc ggtcctattg acgtatatgg cgcctccccc attgacgtca
     5281 attacggtaa atggcccgcc tggctcaatg cccattgacg tcaataggac cacccaccat
     5341 tgacgtcaat gggatggctc attgcccatt catatccgtt ctcacgcccc ctattgacgt
     5401 caatgacggt aaatggccca cttggcagta catcaatatc tattaatagt aacttggcaa
     5461 gtacattact attggaagta cgccagggta cattggcagt actcccattg acgtcaatgg
     5521 cggtaaatgg cccgcgatgg ctgccaagta catccccatt gacgtcaatg gggaggggca
     5581 atgacgcaaa tgggcgttcc attgacgtaa atgggcggta ggcgtgccta atgggaggtc
     5641 tatataagca atgctcgttt agggaaccgc cattctg
//
