LOCUS       sequence-208175-        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: pHBS935 TDP43 RRM-GFP M-S
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..48
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     30..47
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    103..127
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      146..155
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1655..1679)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1724..1858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1748..1767)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1802..1821
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1971..1991)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1971..1989)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2010..2026)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2010..2026)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2023..2045)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2034..2050
                     /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(2058..2064,2065..2082,2083..2088))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2103..2124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2241..2258)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2412..3000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2492..2511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3171..3962,3963..4031))
                     /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     3794..3813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4032..4136)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4162..4617)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4299..4320)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4511..4530
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4688..5672
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 ttctgcctgg ggacgtcgga gcaagcttga tttaggtgac actatagaat acaagctact
       61 tgttcttttt gcaggatccc atcgattcga attcaaggat caacaagttt gtacaaaaaa
      121 gcaggctggc gccggtacca ccggtgccac catgggatcc atgtctgaat atattcgggt
      181 aaccgaagat gagaacgatg agcccattga aataccatcg gaagacgatg ggacggtgct
      241 gctctccacg gttacagccc agtttccagg ggcgtgtggg cttcgctaca ggaatccagt
      301 gtctcagtgt atgagaggtg tccggctggt agaaggaatt ctgcatgccc cagatgctgg
      361 ctggggaaat ctggtgtatg ttgtcaacta tccaaaagat aacaaaagaa aaatggatga
      421 gacagatgct tcatcagcag tgaaagtgaa aagagcagtc cagaaaacat ccagtaaagg
      481 agaggaacta tttacaggta ctgtgcctat caaagtcgag ctggacggcg acgtaaacgg
      541 ccacaagttc agcgtgcgtg gcgagggcga gggcgatgcc accgagggca agctgaccct
      601 gaagttcatc tgcaccaccg gcaagctgcc ggtgccgtgg ccgaccctcg tgaccaccct
      661 gacctacggc gtgcagtgct tcagccgcta cccggaccac atgaagcgtc acgacttctt
      721 caagtccgcc atgccggaag gctacgtcca ggagcgcacc atcgagttca aggacgacgg
      781 cacctacaag acccgcgcgg aggtgaagtt cgagggcgac accctggtga accgcatcga
      841 gctgaagggc aacgacttca aggaggacgg caacatcctg ggtcacaagc tggagtacaa
      901 ccacaacagc cacaacgtcc gtatcgaagc cgacaagcag aagaacggca tcaaggctaa
      961 cttcaagatc cgccacaacg ttgaggacgg cagccagcag gaagccgacc acaaacagca
     1021 gaacaccccg atcggcgacg gcccggtgcg cctgccggac aaccactatc tgagcaccca
     1081 gaccacgctg agcaaagacc cgaacgagaa gcgcgatcac atggtcctga aggagttcgt
     1141 gacggcagcc ggtatcacta aaggtgaaga tgaaagtgga tccaatgccg aacctaaaca
     1201 taatagcaat cgtcagctcg agcggagcgg gcggtttggt ggcaatcctg gtggtttcgg
     1261 caatcaaggt ggatttggaa attcacgcgg tggaggtgca ggactgggca ataaccaggg
     1321 gtccaactcg ggcgggggta gtaactttgg ggcctttagc attaatccgg cgatgatggc
     1381 ggccgcacaa gccgcgctgc aaagcagttg gggtatgatg ggaatgctgg catctcagca
     1441 gaatcagtcc ggtccctctg ggaataatca gaaccagggt aattctcagc gtgagccaaa
     1501 ccaggctttt ggtagcggaa acaattcgta ttcaggcagc aacagcggtg ctgccattgg
     1561 ctggggatct gcctctaatg ctggctcggg atctggtttt aacgggggct tcggcagttc
     1621 atccgattcc aaatcaagtg gatggggatc atagacccag ctttcttgta caaagtggtt
     1681 gatcctctcg agcctctaga actatagtga gtcgtattac gtagatccag acatgataag
     1741 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     1801 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     1861 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     1921 attcgcggcc gcggcgccaa tgcattgggc ccggtaccca gcttttgttc cctttagtga
     1981 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     2041 ccgctcacaa ttccacacaa catacgagcc gggagcataa agtgtaaagc ctggggtgcc
     2101 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     2161 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     2221 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2281 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2341 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2401 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2461 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2521 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2581 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2641 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2701 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2761 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2821 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2881 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2941 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3001 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3061 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     3781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     3841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     3901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     3961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4141 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     4201 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4261 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     4321 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     4381 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     4441 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4501 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     4561 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     4621 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     4681 tacgccagtc gaccgccaat tcaatatggc gtatatggac tcatgccaat tcaatatggt
     4741 ggatctggac ctgtgccaat tcaatatggc gtatatggac tcgtgccaat tcaatatggt
     4801 ggatctggac cccagccaat tcaatatggc ggacttggca ccatgccaat tcaatatggc
     4861 ggacctggca ctgtgccaac tggggagggg tctacttggc acggtgccaa gtttgaggag
     4921 gggtcttggc cctgtgccaa gtccgccata ttgaattggc atggtgccaa taatggcggc
     4981 catattggct atatgccagg atcaatatat aggcaatatc caatatggcc ctatgccaat
     5041 atggctattg gccaggttca atactatgta ttggccctat gccatatagt attccatata
     5101 tgggttttcc tattgacgta gatagcccct cccaatgggc ggtcccatat accatatatg
     5161 gggcttccta ataccgccca tagccactcc cccattgacg tcaatggtct ctatatatgg
     5221 tctttcctat tgacgtcata tgggcggtcc tattgacgta tatggcgcct cccccattga
     5281 cgtcaattac ggtaaatggc ccgcctggct caatgcccat tgacgtcaat aggaccaccc
     5341 accattgacg tcaatgggat ggctcattgc ccattcatat ccgttctcac gccccctatt
     5401 gacgtcaatg acggtaaatg gcccacttgg cagtacatca atatctatta atagtaactt
     5461 ggcaagtaca ttactattgg aagtacgcca gggtacattg gcagtactcc cattgacgtc
     5521 aatggcggta aatggcccgc gatggctgcc aagtacatcc ccattgacgt caatggggag
     5581 gggcaatgac gcaaatgggc gttccattga cgtaaatggg cggtaggcgt gcctaatggg
     5641 aggtctatat aagcaatgct cgtttaggga accgcca
//
