LOCUS       sequence-207921-        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: pHBS1256 TDP43 RRM-GFP W385G
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        44..62
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     44..61
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    117..141
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      160..169
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1669..1693)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1738..1872
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1762..1781)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1816..1835
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1985..2005)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1985..2003)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2024..2040)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2024..2040)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2037..2059)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2048..2064
                     /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(2072..2078,2079..2096,2097..2102))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2117..2138
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2255..2272)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2426..3014)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2506..2525)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3185..3976,3977..4045))
                     /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     3808..3827
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4046..4150)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4176..4631)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4313..4334)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4525..4544
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(4702..5677,1..9)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 ttagggaacc gccattctgc ctggggacgt cggagcaagc ttgatttagg tgacactata
       61 gaatacaagc tacttgttct ttttgcagga tcccatcgat tcgaattcaa ggatcaacaa
      121 gtttgtacaa aaaagcaggc tggcgccggt accaccggtg ccaccatggg atccatgtct
      181 gaatatattc gggtaaccga agatgagaac gatgagccca ttgaaatacc atcggaagac
      241 gatgggacgg tgctgctctc cacggttaca gcccagtttc caggggcgtg tgggcttcgc
      301 tacaggaatc cagtgtctca gtgtatgaga ggtgtccggc tggtagaagg aattctgcat
      361 gccccagatg ctggctgggg aaatctggtg tatgttgtca actatccaaa agataacaaa
      421 agaaaaatgg atgagacaga tgcttcatca gcagtgaaag tgaaaagagc agtccagaaa
      481 acatccagta aaggagagga actatttaca ggtactgtgc ctatcaaagt cgagctggac
      541 ggcgacgtaa acggccacaa gttcagcgtg cgtggcgagg gcgagggcga tgccaccgag
      601 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgccggtgcc gtggccgacc
      661 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctacccgga ccacatgaag
      721 cgtcacgact tcttcaagtc cgccatgccg gaaggctacg tccaggagcg caccatcgag
      781 ttcaaggacg acggcaccta caagacccgc gcggaggtga agttcgaggg cgacaccctg
      841 gtgaaccgca tcgagctgaa gggcaacgac ttcaaggagg acggcaacat cctgggtcac
      901 aagctggagt acaaccacaa cagccacaac gtccgtatcg aagccgacaa gcagaagaac
      961 ggcatcaagg ctaacttcaa gatccgccac aacgttgagg acggcagcca gcaggaagcc
     1021 gaccacaaac agcagaacac cccgatcggc gacggcccgg tgcgcctgcc ggacaaccac
     1081 tatctgagca cccagaccac gctgagcaaa gacccgaacg agaagcgcga tcacatggtc
     1141 ctgaaggagt tcgtgacggc agccggtatc actaaaggtg aagatgaaag tggatccaat
     1201 gccgaaccta agcacaatag caatagacag ttagaaagaa gtggaagatt tggtggtaat
     1261 ccaggtggct ttgggaatca gggtggattt ggtaatagca gagggggtgg agctggtttg
     1321 ggaaacaatc aaggtagtaa tatgggtggt gggatgaact ttggtgcgtt cagcattaat
     1381 ccagccatga tggctgccgc ccaggcagca ctacagagca gttggggtat gatgggcatg
     1441 ttagccagcc agcagaacca gtcaggccca tcgggtaata accaaaacca aggcaacatg
     1501 cagagggagc caaaccaggc cttcggttct ggaaataact cttatagtgg ctctaattct
     1561 ggtgcagcaa ttggtggggg atcagcatcc aatgcagggt cgggcagtgg ttttaatgga
     1621 ggctttggct caagcatgga ttctaagtct tctggctggg gaatgtagac ccagctttct
     1681 tgtacaaagt ggttgatcct ctcgagcctc tagaactata gtgagtcgta ttacgtagat
     1741 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     1801 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     1861 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     1921 tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt
     1981 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
     2041 tgtgaaattg ttatccgctc acaattccac acaacatacg agccgggagc ataaagtgta
     2101 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2161 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     2221 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2281 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2341 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2401 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2461 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2521 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2581 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2641 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2701 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2761 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2821 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2881 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2941 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3001 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3061 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3121 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3181 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3241 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3301 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3361 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     3421 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     3481 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     3541 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     3601 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     3661 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     3721 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     3781 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     3841 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     3901 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     3961 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4021 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4081 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4141 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     4201 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     4261 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     4321 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     4381 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     4441 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     4501 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     4561 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     4621 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     4681 ggcctcttcg ctattacgcc agtcgaccgc caattcaata tggcgtatat ggactcatgc
     4741 caattcaata tggtggatct ggacctgtgc caattcaata tggcgtatat ggactcgtgc
     4801 caattcaata tggtggatct ggaccccagc caattcaata tggcggactt ggcaccatgc
     4861 caattcaata tggcggacct ggcactgtgc caactgggga ggggtctact tggcacggtg
     4921 ccaagtttga ggaggggtct tggccctgtg ccaagtccgc catattgaat tggcatggtg
     4981 ccaataatgg cggccatatt ggctatatgc caggatcaat atataggcaa tatccaatat
     5041 ggccctatgc caatatggct attggccagg ttcaatacta tgtattggcc ctatgccata
     5101 tagtattcca tatatgggtt ttcctattga cgtagatagc ccctcccaat gggcggtccc
     5161 atataccata tatggggctt cctaataccg cccatagcca ctcccccatt gacgtcaatg
     5221 gtctctatat atggtctttc ctattgacgt catatgggcg gtcctattga cgtatatggc
     5281 gcctccccca ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt
     5341 caataggacc acccaccatt gacgtcaatg ggatggctca ttgcccattc atatccgttc
     5401 tcacgccccc tattgacgtc aatgacggta aatggcccac ttggcagtac atcaatatct
     5461 attaatagta acttggcaag tacattacta ttggaagtac gccagggtac attggcagta
     5521 ctcccattga cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac atccccattg
     5581 acgtcaatgg ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag
     5641 gcgtgcctaa tgggaggtct atataagcaa tgctcgt
//
