LOCUS       sequence-207993-        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: pHBS1292 TDP43 RRM-GFP G368W+W385G
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        34..52
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     34..51
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    107..131
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      150..159
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1659..1683)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1728..1862
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1752..1771)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1806..1825
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1975..1995)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1975..1993)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2014..2030)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2014..2030)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2027..2049)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2038..2054
                     /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(2062..2068,2069..2086,2087..2092))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2107..2128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2245..2262)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2416..3004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2496..2515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3175..3966,3967..4035))
                     /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     3798..3817
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4036..4140)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4166..4621)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4303..4324)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4515..4534
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4692..5676
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 gccattctgc ctggggacgt cggagcaagc ttgatttagg tgacactata gaatacaagc
       61 tacttgttct ttttgcagga tcccatcgat tcgaattcaa ggatcaacaa gtttgtacaa
      121 aaaagcaggc tggcgccggt accaccggtg ccaccatggg atccatgtct gaatatattc
      181 gggtaaccga agatgagaac gatgagccca ttgaaatacc atcggaagac gatgggacgg
      241 tgctgctctc cacggttaca gcccagtttc caggggcgtg tgggcttcgc tacaggaatc
      301 cagtgtctca gtgtatgaga ggtgtccggc tggtagaagg aattctgcat gccccagatg
      361 ctggctgggg aaatctggtg tatgttgtca actatccaaa agataacaaa agaaaaatgg
      421 atgagacaga tgcttcatca gcagtgaaag tgaaaagagc agtccagaaa acatccagta
      481 aaggagagga actatttaca ggtactgtgc ctatcaaagt cgagctggac ggcgacgtaa
      541 acggccacaa gttcagcgtg cgtggcgagg gcgagggcga tgccaccgag ggcaagctga
      601 ccctgaagtt catctgcacc accggcaagc tgccggtgcc gtggccgacc ctcgtgacca
      661 ccctgaccta cggcgtgcag tgcttcagcc gctacccgga ccacatgaag cgtcacgact
      721 tcttcaagtc cgccatgccg gaaggctacg tccaggagcg caccatcgag ttcaaggacg
      781 acggcaccta caagacccgc gcggaggtga agttcgaggg cgacaccctg gtgaaccgca
      841 tcgagctgaa gggcaacgac ttcaaggagg acggcaacat cctgggtcac aagctggagt
      901 acaaccacaa cagccacaac gtccgtatcg aagccgacaa gcagaagaac ggcatcaagg
      961 ctaacttcaa gatccgccac aacgttgagg acggcagcca gcaggaagcc gaccacaaac
     1021 agcagaacac cccgatcggc gacggcccgg tgcgcctgcc ggacaaccac tatctgagca
     1081 cccagaccac gctgagcaaa gacccgaacg agaagcgcga tcacatggtc ctgaaggagt
     1141 tcgtgacggc agccggtatc actaaaggtg aagatgaaag tggatccaat gccgaaccta
     1201 agcacaatag caatagacag ttagaaagaa gtggaagatt tggtggtaat ccaggtggct
     1261 ttgggaatca gggtggattt ggtaatagca gagggggtgg agctggtttg ggaaacaatc
     1321 aaggtagtaa tatgggtggt gggatgaact ttggtgcgtt cagcattaat ccagccatga
     1381 tggctgccgc ccaggcagca ctacagagca gttggggtat gatgggcatg ttagccagcc
     1441 agcagaacca gtcaggccca tcgggtaata accaaaacca aggcaacatg cagagggagc
     1501 caaaccaggc cttctggtct ggaaataact cttatagtgg ctctaattct ggtgcagcaa
     1561 ttggtggggg atcagcatcc aatgcagggt cgggcagtgg ttttaatgga ggctttggct
     1621 caagcatgga ttctaagtct tctggctggg gaatgtagac ccagctttct tgtacaaagt
     1681 ggttgatcct ctcgagcctc tagaactata gtgagtcgta ttacgtagat ccagacatga
     1741 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     1801 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     1861 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     1921 tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta
     1981 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     2041 ttatccgctc acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg
     2101 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2161 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2221 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2281 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2341 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2401 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2461 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2521 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2581 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2641 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2701 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2761 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2821 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2881 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2941 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3001 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3061 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3121 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3181 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3241 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3301 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3361 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3421 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3481 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3541 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3601 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3661 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3721 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3781 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3841 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3901 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3961 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4021 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4081 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4141 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     4201 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     4261 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     4321 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     4381 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     4441 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4501 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     4561 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     4621 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     4681 ctattacgcc agtcgaccgc caattcaata tggcgtatat ggactcatgc caattcaata
     4741 tggtggatct ggacctgtgc caattcaata tggcgtatat ggactcgtgc caattcaata
     4801 tggtggatct ggaccccagc caattcaata tggcggactt ggcaccatgc caattcaata
     4861 tggcggacct ggcactgtgc caactgggga ggggtctact tggcacggtg ccaagtttga
     4921 ggaggggtct tggccctgtg ccaagtccgc catattgaat tggcatggtg ccaataatgg
     4981 cggccatatt ggctatatgc caggatcaat atataggcaa tatccaatat ggccctatgc
     5041 caatatggct attggccagg ttcaatacta tgtattggcc ctatgccata tagtattcca
     5101 tatatgggtt ttcctattga cgtagatagc ccctcccaat gggcggtccc atataccata
     5161 tatggggctt cctaataccg cccatagcca ctcccccatt gacgtcaatg gtctctatat
     5221 atggtctttc ctattgacgt catatgggcg gtcctattga cgtatatggc gcctccccca
     5281 ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt caataggacc
     5341 acccaccatt gacgtcaatg ggatggctca ttgcccattc atatccgttc tcacgccccc
     5401 tattgacgtc aatgacggta aatggcccac ttggcagtac atcaatatct attaatagta
     5461 acttggcaag tacattacta ttggaagtac gccagggtac attggcagta ctcccattga
     5521 cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac atccccattg acgtcaatgg
     5581 ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa
     5641 tgggaggtct atataagcaa tgctcgttta gggaacc
//
