LOCUS       sequence-208171-        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: pHBS931 TDP43 RRM-GFP F-S
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..42
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     24..41
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    97..121
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      140..149
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1649..1673)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1718..1852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1742..1761)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1796..1815
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1965..1985)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1965..1983)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2004..2020)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2004..2020)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2017..2039)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2028..2044
                     /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(2052..2058,2059..2076,2077..2082))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2097..2118
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2235..2252)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2406..2994)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2486..2505)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3165..3956,3957..4025))
                     /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     3788..3807
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4026..4130)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4156..4611)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4293..4314)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4505..4524
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4682..5666
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 ctggggacgt cggagcaagc ttgatttagg tgacactata gaatacaagc tacttgttct
       61 ttttgcagga tcccatcgat tcgaattcaa ggatcaacaa gtttgtacaa aaaagcaggc
      121 tggcgccggt accaccggtg ccaccatggg atccatgtct gaatatattc gggtaaccga
      181 agatgagaac gatgagccca ttgaaatacc atcggaagac gatgggacgg tgctgctctc
      241 cacggttaca gcccagtttc caggggcgtg tgggcttcgc tacaggaatc cagtgtctca
      301 gtgtatgaga ggtgtccggc tggtagaagg aattctgcat gccccagatg ctggctgggg
      361 aaatctggtg tatgttgtca actatccaaa agataacaaa agaaaaatgg atgagacaga
      421 tgcttcatca gcagtgaaag tgaaaagagc agtccagaaa acatccagta aaggagagga
      481 actatttaca ggtactgtgc ctatcaaagt cgagctggac ggcgacgtaa acggccacaa
      541 gttcagcgtg cgtggcgagg gcgagggcga tgccaccgag ggcaagctga ccctgaagtt
      601 catctgcacc accggcaagc tgccggtgcc gtggccgacc ctcgtgacca ccctgaccta
      661 cggcgtgcag tgcttcagcc gctacccgga ccacatgaag cgtcacgact tcttcaagtc
      721 cgccatgccg gaaggctacg tccaggagcg caccatcgag ttcaaggacg acggcaccta
      781 caagacccgc gcggaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
      841 gggcaacgac ttcaaggagg acggcaacat cctgggtcac aagctggagt acaaccacaa
      901 cagccacaac gtccgtatcg aagccgacaa gcagaagaac ggcatcaagg ctaacttcaa
      961 gatccgccac aacgttgagg acggcagcca gcaggaagcc gaccacaaac agcagaacac
     1021 cccgatcggc gacggcccgg tgcgcctgcc ggacaaccac tatctgagca cccagaccac
     1081 gctgagcaaa gacccgaacg agaagcgcga tcacatggtc ctgaaggagt tcgtgacggc
     1141 agccggtatc actaaaggtg aagatgaaag tggatccaat gccgaaccta aacataattc
     1201 aaaccggcag ttggaaagat cggggcgcag tggtggcaac ccaggcggtt ctggtaatca
     1261 gggcggctcg ggaaattcca gaggcggggg tgcagggctt ggcaacaatc aggggtccaa
     1321 tatgggcggt ggtatgaact caggtgccag ctcgattaat ccggccatga tggcggcagc
     1381 gcaggcggcg ctgcagtctt cctggggtat gatgggtatg ttagcatccc aacagaacca
     1441 gagtggcccg tcggggaata accaaaatca ggggaatatg cagcgcgaac cgaatcaagc
     1501 ttcaggtagt ggtaacaaca gttatagtgg atcgaacagt ggcgccgcaa ttggatgggg
     1561 ttccgcatcc aatgcaggct caggcagcgg ctcgaatggc ggctcaggtt ctagtatgga
     1621 ttcgaagagc tctggttggg gaatgtagac ccagctttct tgtacaaagt ggttgatcct
     1681 ctcgagcctc tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat
     1741 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     1801 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     1861 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc
     1921 ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta
     1981 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     2041 acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga
     2101 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2161 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2221 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2281 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2341 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2401 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2461 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2521 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2581 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2641 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2701 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2761 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2821 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2881 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2941 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3001 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3061 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3121 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3181 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3241 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3301 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3361 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3421 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3481 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3541 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3601 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3661 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3721 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3781 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3841 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3901 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3961 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4021 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4081 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4141 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     4201 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4261 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4321 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4381 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4441 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4501 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     4561 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     4621 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     4681 agtcgaccgc caattcaata tggcgtatat ggactcatgc caattcaata tggtggatct
     4741 ggacctgtgc caattcaata tggcgtatat ggactcgtgc caattcaata tggtggatct
     4801 ggaccccagc caattcaata tggcggactt ggcaccatgc caattcaata tggcggacct
     4861 ggcactgtgc caactgggga ggggtctact tggcacggtg ccaagtttga ggaggggtct
     4921 tggccctgtg ccaagtccgc catattgaat tggcatggtg ccaataatgg cggccatatt
     4981 ggctatatgc caggatcaat atataggcaa tatccaatat ggccctatgc caatatggct
     5041 attggccagg ttcaatacta tgtattggcc ctatgccata tagtattcca tatatgggtt
     5101 ttcctattga cgtagatagc ccctcccaat gggcggtccc atataccata tatggggctt
     5161 cctaataccg cccatagcca ctcccccatt gacgtcaatg gtctctatat atggtctttc
     5221 ctattgacgt catatgggcg gtcctattga cgtatatggc gcctccccca ttgacgtcaa
     5281 ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt caataggacc acccaccatt
     5341 gacgtcaatg ggatggctca ttgcccattc atatccgttc tcacgccccc tattgacgtc
     5401 aatgacggta aatggcccac ttggcagtac atcaatatct attaatagta acttggcaag
     5461 tacattacta ttggaagtac gccagggtac attggcagta ctcccattga cgtcaatggc
     5521 ggtaaatggc ccgcgatggc tgccaagtac atccccattg acgtcaatgg ggaggggcaa
     5581 tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa tgggaggtct
     5641 atataagcaa tgctcgttta gggaaccgcc attctgc
//
