LOCUS       sequence-229822-        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: pHBS1386 TDP43 RRM-GFP VLIM-F
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        45..1022
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1057..1075
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1057..1074
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1130..1154
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1173..1182
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2682..2706)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2751..2885
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2775..2794)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2829..2848
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2998..3018)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2998..3016)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3037..3053)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3037..3053)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3050..3072)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3061..3077
                     /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(3085..3091,3092..3109,3110..3115))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3130..3151
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3268..3285)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3439..4027)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3519..3538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4198..4989,4990..5058))
                     /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     4821..4840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5059..5163)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5189..5644)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5326..5347)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5538..5557
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 aagggcgatc ggtgcgggcc tcttcgctat tacgccagtc gaccgccaat tcaatatggc
       61 gtatatggac tcatgccaat tcaatatggt ggatctggac ctgtgccaat tcaatatggc
      121 gtatatggac tcgtgccaat tcaatatggt ggatctggac cccagccaat tcaatatggc
      181 ggacttggca ccatgccaat tcaatatggc ggacctggca ctgtgccaac tggggagggg
      241 tctacttggc acggtgccaa gtttgaggag gggtcttggc cctgtgccaa gtccgccata
      301 ttgaattggc atggtgccaa taatggcggc catattggct atatgccagg atcaatatat
      361 aggcaatatc caatatggcc ctatgccaat atggctattg gccaggttca atactatgta
      421 ttggccctat gccatatagt attccatata tgggttttcc tattgacgta gatagcccct
      481 cccaatgggc ggtcccatat accatatatg gggcttccta ataccgccca tagccactcc
      541 cccattgacg tcaatggtct ctatatatgg tctttcctat tgacgtcata tgggcggtcc
      601 tattgacgta tatggcgcct cccccattga cgtcaattac ggtaaatggc ccgcctggct
      661 caatgcccat tgacgtcaat aggaccaccc accattgacg tcaatgggat ggctcattgc
      721 ccattcatat ccgttctcac gccccctatt gacgtcaatg acggtaaatg gcccacttgg
      781 cagtacatca atatctatta atagtaactt ggcaagtaca ttactattgg aagtacgcca
      841 gggtacattg gcagtactcc cattgacgtc aatggcggta aatggcccgc gatggctgcc
      901 aagtacatcc ccattgacgt caatggggag gggcaatgac gcaaatgggc gttccattga
      961 cgtaaatggg cggtaggcgt gcctaatggg aggtctatat aagcaatgct cgtttaggga
     1021 accgccattc tgcctgggga cgtcggagca agcttgattt aggtgacact atagaataca
     1081 agctacttgt tctttttgca ggatcccatc gattcgaatt caaggatcaa caagtttgta
     1141 caaaaaagca ggctggcgcc ggtaccaccg gtgccaccat gggatccatg tctgaatata
     1201 ttcgggtaac cgaagatgag aacgatgagc ccattgaaat accatcggaa gacgatggga
     1261 cggtgctgct ctccacggtt acagcccagt ttccaggggc gtgtgggctt cgctacagga
     1321 atccagtgtc tcagtgtatg agaggtgtcc ggctggtaga aggaattctg catgccccag
     1381 atgctggctg gggaaatctg gtgtatgttg tcaactatcc aaaagataac aaaagaaaaa
     1441 tggatgagac agatgcttca tcagcagtga aagtgaaaag agcagtccag aaaacatcca
     1501 gtaaaggaga ggaactattt acaggtactg tgcctatcaa agtcgagctg gacggcgacg
     1561 taaacggcca caagttcagc gtgcgtggcg agggcgaggg cgatgccacc gagggcaagc
     1621 tgaccctgaa gttcatctgc accaccggca agctgccggt gccgtggccg accctcgtga
     1681 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc ggaccacatg aagcgtcacg
     1741 acttcttcaa gtccgccatg ccggaaggct acgtccagga gcgcaccatc gagttcaagg
     1801 acgacggcac ctacaagacc cgcgcggagg tgaagttcga gggcgacacc ctggtgaacc
     1861 gcatcgagct gaagggcaac gacttcaagg aggacggcaa catcctgggt cacaagctgg
     1921 agtacaacca caacagccac aacgtccgta tcgaagccga caagcagaag aacggcatca
     1981 aggctaactt caagatccgc cacaacgttg aggacggcag ccagcaggaa gccgaccaca
     2041 aacagcagaa caccccgatc ggcgacggcc cggtgcgcct gccggacaac cactatctga
     2101 gcacccagac cacgctgagc aaagacccga acgagaagcg cgatcacatg gtcctgaagg
     2161 agttcgtgac ggcagccggt atcactaaag gtgaagatga aagtggatcc aatgccgaac
     2221 ctaagcataa cagtaatcgg caatttgagc gatccggcag atttggaggg aaccctggag
     2281 gattcggcaa ccaggggggc tttggaaatt caagaggagg gggtgcggga ttcggtaaca
     2341 accaggggtc caatttcggt ggggggttca actttggggc gttttctttc aatcccgcta
     2401 tgatggctgc ggctcaagcg gctctgcaaa gtagctgggg catgatgggt atgctcgcgt
     2461 cacaacagaa tcagagtgga ccaagtggaa ataatcagaa tcaagggaat tttcaacgag
     2521 aacctaacca agcattcgga tctggtaata atagctactc cgggagtaac tccggagcag
     2581 ctttcgggtg gggctccgcg tcaaacgctg gcagcggctc tggattcaac gggggattcg
     2641 gaagcagttt cgattcaaaa agcagtggct gggggttcta gacccagctt tcttgtacaa
     2701 agtggttgat cctctcgagc ctctagaact atagtgagtc gtattacgta gatccagaca
     2761 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     2821 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     2881 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     2941 ttttttaatt cgcggccgcg gcgccaatgc attgggcccg gtacccagct tttgttccct
     3001 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3061 ttgttatccg ctcacaattc cacacaacat acgagccggg agcataaagt gtaaagcctg
     3121 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3181 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3241 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3301 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3361 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3421 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3481 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3541 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3601 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3661 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3721 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3781 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3841 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3901 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3961 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4021 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4081 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4141 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4201 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4261 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4321 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4381 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4441 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4501 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4561 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4621 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4681 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4741 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4801 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4861 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4921 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4981 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5041 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5101 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5161 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     5221 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     5281 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     5341 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     5401 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     5461 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     5521 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     5581 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     5641 gcgtcccatt cgccattcag gctgcgcaac tgttggg
//
