LOCUS       sequence-229819-        5677 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To test the effect of sequence on TDP43 RRM-GFP 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: pHBS1388 TDP43 RRM-GFP K-R
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        19..996
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1031..1049
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1031..1048
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1104..1128
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1147..1156
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2656..2680)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2725..2859
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2749..2768)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2803..2822
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2972..2992)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2972..2990)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3011..3027)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3011..3027)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3024..3046)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3035..3051
                     /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(3059..3065,3066..3083,3084..3089))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3104..3125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3242..3259)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3413..4001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3493..3512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4172..4963,4964..5032))
                     /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     4795..4814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5033..5137)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5163..5618)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5300..5321)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5512..5531
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctattacgcc agtcgaccgc caattcaata tggcgtatat ggactcatgc caattcaata
       61 tggtggatct ggacctgtgc caattcaata tggcgtatat ggactcgtgc caattcaata
      121 tggtggatct ggaccccagc caattcaata tggcggactt ggcaccatgc caattcaata
      181 tggcggacct ggcactgtgc caactgggga ggggtctact tggcacggtg ccaagtttga
      241 ggaggggtct tggccctgtg ccaagtccgc catattgaat tggcatggtg ccaataatgg
      301 cggccatatt ggctatatgc caggatcaat atataggcaa tatccaatat ggccctatgc
      361 caatatggct attggccagg ttcaatacta tgtattggcc ctatgccata tagtattcca
      421 tatatgggtt ttcctattga cgtagatagc ccctcccaat gggcggtccc atataccata
      481 tatggggctt cctaataccg cccatagcca ctcccccatt gacgtcaatg gtctctatat
      541 atggtctttc ctattgacgt catatgggcg gtcctattga cgtatatggc gcctccccca
      601 ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt caataggacc
      661 acccaccatt gacgtcaatg ggatggctca ttgcccattc atatccgttc tcacgccccc
      721 tattgacgtc aatgacggta aatggcccac ttggcagtac atcaatatct attaatagta
      781 acttggcaag tacattacta ttggaagtac gccagggtac attggcagta ctcccattga
      841 cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac atccccattg acgtcaatgg
      901 ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa
      961 tgggaggtct atataagcaa tgctcgttta gggaaccgcc attctgcctg gggacgtcgg
     1021 agcaagcttg atttaggtga cactatagaa tacaagctac ttgttctttt tgcaggatcc
     1081 catcgattcg aattcaagga tcaacaagtt tgtacaaaaa agcaggctgg cgccggtacc
     1141 accggtgcca ccatgggatc catgtctgaa tatattcggg taaccgaaga tgagaacgat
     1201 gagcccattg aaataccatc ggaagacgat gggacggtgc tgctctccac ggttacagcc
     1261 cagtttccag gggcgtgtgg gcttcgctac aggaatccag tgtctcagtg tatgagaggt
     1321 gtccggctgg tagaaggaat tctgcatgcc ccagatgctg gctggggaaa tctggtgtat
     1381 gttgtcaact atccaaaaga taacaaaaga aaaatggatg agacagatgc ttcatcagca
     1441 gtgaaagtga aaagagcagt ccagaaaaca tccagtaaag gagaggaact atttacaggt
     1501 actgtgccta tcaaagtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgcgt
     1561 ggcgagggcg agggcgatgc caccgagggc aagctgaccc tgaagttcat ctgcaccacc
     1621 ggcaagctgc cggtgccgtg gccgaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     1681 ttcagccgct acccggacca catgaagcgt cacgacttct tcaagtccgc catgccggaa
     1741 ggctacgtcc aggagcgcac catcgagttc aaggacgacg gcacctacaa gacccgcgcg
     1801 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg caacgacttc
     1861 aaggaggacg gcaacatcct gggtcacaag ctggagtaca accacaacag ccacaacgtc
     1921 cgtatcgaag ccgacaagca gaagaacggc atcaaggcta acttcaagat ccgccacaac
     1981 gttgaggacg gcagccagca ggaagccgac cacaaacagc agaacacccc gatcggcgac
     2041 ggcccggtgc gcctgccgga caaccactat ctgagcaccc agaccacgct gagcaaagac
     2101 ccgaacgaga agcgcgatca catggtcctg aaggagttcg tgacggcagc cggtatcact
     2161 aaaggtgaag atgaaagtgg atccaatgcc gaacctaggc ataactccaa tcggcaactt
     2221 gagcgcagtg ggaggttcgg cgggaatcca ggcggttttg gtaatcaagg agggtttggt
     2281 aacagtaggg gtggaggagc aggactgggg aataaccagg gtagtaatat gggcggtgga
     2341 atgaatttcg gtgccttttc tattaaccca gccatgatgg cagcagcaca ggcggctctt
     2401 cagtcatcct ggggtatgat gggtatgttg gcaagccaac agaatcaatc aggaccgtcc
     2461 ggtaacaacc aaaatcaggg aaacatgcag cgggaaccga accaagcctt cggtagcggt
     2521 aacaattcct atagcggtag taactcaggc gcggcgatcg gctggggctc agcgtcaaat
     2581 gcgggatctg ggagcgggtt caacggaggt ttcggttcca gcatggacag tcggtcctca
     2641 ggatggggta tgtagaccca gctttcttgt acaaagtggt tgatcctctc gagcctctag
     2701 aactatagtg agtcgtatta cgtagatcca gacatgataa gatacattga tgagtttgga
     2761 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2821 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     2881 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aattcgcggc cgcggcgcca
     2941 atgcattggg cccggtaccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     3001 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3061 acatacgagc cgggagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3121 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3181 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3241 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3301 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3361 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3421 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3481 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3541 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3601 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3661 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3721 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3781 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3841 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3901 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3961 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4021 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4081 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4141 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4201 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4261 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4321 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4381 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4441 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4501 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4561 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4621 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4681 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4741 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4801 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4861 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4921 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4981 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5041 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5101 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5161 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     5221 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     5281 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     5341 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     5401 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     5461 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     5521 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     5581 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
     5641 caactgttgg gaagggcgat cggtgcgggc ctcttcg
//
