LOCUS       sequence-207949-        5682 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 5682)
  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 5682)
  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: pHBS968 TDP43 RRM-GFP G309S
FEATURES             Location/Qualifiers
     source          1..5682
                     /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(1654..1678)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1723..1857
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1747..1766)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1801..1820
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1970..1990)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1970..1988)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2009..2025)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2009..2025)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2022..2044)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2033..2049
                     /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(2057..2063,2064..2081,2082..2087))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2102..2123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2240..2257)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2411..2999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2491..2510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3170..3961,3962..4030))
                     /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     3793..3812
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4031..4135)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4161..4616)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4298..4319)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4510..4529
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4687..5671
                     /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 agcacaatag
     1201 caatagacag ttagaaagaa gtggaagatt tggtggtaat ccaggtggct ttgggaatca
     1261 gggtggattt ggtaatagca gagggggtgg agctggtttg ggaaacaatc aaggtagtaa
     1321 tatgggtagt gggatgaact ttggtgcgtt cagcattaat ccagccatga tggctgccgc
     1381 ccaggcagca ctacagagca gttggggtat gatgggcatg ttagccagcc agcagaacca
     1441 gtcaggccca tcgggtaata accaaaacca aggcaacatg cagagggagc caaaccaggc
     1501 cttcggttct ggaaataact cttatagtgg ctctaattct ggtgcagcaa ttggttgggg
     1561 atcagcatcc aatgcagggt cgggcagtgg ttttaatgga ggctttggct caagcatgga
     1621 ttctaagtct tctggctggg gaatgtagac tagacccagc tttcttgtac aaagtggttg
     1681 atcctctcga gcctctagaa ctatagtgag tcgtattacg tagatccaga catgataaga
     1741 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     1801 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     1861 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     1921 ttcgcggccg cggcgccaat gcattgggcc cggtacccag cttttgttcc ctttagtgag
     1981 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2041 cgctcacaat tccacacaac atacgagccg ggagcataaa gtgtaaagcc tggggtgcct
     2101 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2161 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2221 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2281 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2341 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2401 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2461 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2521 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2581 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2641 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2701 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2761 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2821 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2881 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2941 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3001 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3061 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3121 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3181 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3241 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3301 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3361 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3421 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3481 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3541 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3601 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3661 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3721 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3781 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3841 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3901 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3961 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4021 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4081 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4141 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     4201 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     4261 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     4321 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     4381 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     4441 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     4501 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     4561 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
     4621 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     4681 acgccagtcg accgccaatt caatatggcg tatatggact catgccaatt caatatggtg
     4741 gatctggacc tgtgccaatt caatatggcg tatatggact cgtgccaatt caatatggtg
     4801 gatctggacc ccagccaatt caatatggcg gacttggcac catgccaatt caatatggcg
     4861 gacctggcac tgtgccaact ggggaggggt ctacttggca cggtgccaag tttgaggagg
     4921 ggtcttggcc ctgtgccaag tccgccatat tgaattggca tggtgccaat aatggcggcc
     4981 atattggcta tatgccagga tcaatatata ggcaatatcc aatatggccc tatgccaata
     5041 tggctattgg ccaggttcaa tactatgtat tggccctatg ccatatagta ttccatatat
     5101 gggttttcct attgacgtag atagcccctc ccaatgggcg gtcccatata ccatatatgg
     5161 ggcttcctaa taccgcccat agccactccc ccattgacgt caatggtctc tatatatggt
     5221 ctttcctatt gacgtcatat gggcggtcct attgacgtat atggcgcctc ccccattgac
     5281 gtcaattacg gtaaatggcc cgcctggctc aatgcccatt gacgtcaata ggaccaccca
     5341 ccattgacgt caatgggatg gctcattgcc cattcatatc cgttctcacg ccccctattg
     5401 acgtcaatga cggtaaatgg cccacttggc agtacatcaa tatctattaa tagtaacttg
     5461 gcaagtacat tactattgga agtacgccag ggtacattgg cagtactccc attgacgtca
     5521 atggcggtaa atggcccgcg atggctgcca agtacatccc cattgacgtc aatggggagg
     5581 ggcaatgacg caaatgggcg ttccattgac gtaaatgggc ggtaggcgtg cctaatggga
     5641 ggtctatata agcaatgctc gtttagggaa ccgccattct gc
//
