LOCUS       sequence-235275-        5677 bp    DNA     circular SYN 15-APR-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 Apr 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: pHBS941 TDP43 RRM-GFP VLIMFYW-S
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        43..1027
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1062..1080
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1062..1079
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1135..1159
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1178..1187
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2687..2711)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2756..2890
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2780..2799)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2834..2853
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3003..3023)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3003..3021)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3042..3058)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3042..3058)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3055..3077)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3066..3082
                     /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(3090..3096,3097..3114,3115..3120))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3135..3156
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3273..3290)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3444..4032)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3524..3543)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4203..4994,4995..5063))
                     /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     4826..4845
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5064..5168)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5194..5649)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5331..5352)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5543..5562
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgaccg ccaattcaat
       61 atggcgtata tggactcatg ccaattcaat atggtggatc tggacctgtg ccaattcaat
      121 atggcgtata tggactcgtg ccaattcaat atggtggatc tggaccccag ccaattcaat
      181 atggcggact tggcaccatg ccaattcaat atggcggacc tggcactgtg ccaactgggg
      241 aggggtctac ttggcacggt gccaagtttg aggaggggtc ttggccctgt gccaagtccg
      301 ccatattgaa ttggcatggt gccaataatg gcggccatat tggctatatg ccaggatcaa
      361 tatataggca atatccaata tggccctatg ccaatatggc tattggccag gttcaatact
      421 atgtattgac cctatgccat atagtattcc atatatgggt tttcctattg acgtagatag
      481 cccctcccaa tgggcggtcc catataccat atatggggct tcctaatacc gcccatagcc
      541 actcccccat tgacgtcaat ggtctctata tatggtcttt cctattgacg tcatatgggc
      601 ggtcctattg acgtatatgg cgcctccccc attgacgtca attacggtaa atggcccgcc
      661 tggctcaatg cccattgacg tcaataggac cacccaccat tgacgtcaat gggatggctc
      721 attgcccatt catatccgtt ctcacgcccc ctattgacgt caatgacggt aaatggccca
      781 cttggcagta catcaatatc tattaatagt aacttggcaa gtacattact attggaagta
      841 cgccagggta cattggcagt actcccattg acgtcaatgg cggtaaatgg cccgcgatgg
      901 ctgccaagta catccccatt gacgtcaatg gggaggggca atgacgcaaa tgggcgttcc
      961 attgacgtaa atgggcggta ggcgtgccta atgggaggtc tatataagca atgctcgttt
     1021 agggaaccgc cattctgcct ggggacgtcg gagcaagctt gatttaggtg acactataga
     1081 atacaagcta cttgttcttt ttgcaggatc ccatcgattc gaattcaagg atcaacaagt
     1141 ttgtacaaaa aagcaggctg gcgccggtac caccggtgcc accatgggat ccatgtctga
     1201 atatattcgg gtaaccgaag atgagaacga tgagcccatt gaaataccat cggaagacga
     1261 tgggacggtg ctgctctcca cggttacagc ccagtttcca ggggcgtgtg ggcttcgcta
     1321 caggaatcca gtgtctcagt gtatgagagg tgtccggctg gtagaaggaa ttctgcatgc
     1381 cccagatgct ggctggggaa atctggtgta tgttgtcaac tatccaaaag ataacaaaag
     1441 aaaaatggat gagacagatg cttcatcagc agtgaaagtg aaaagagcag tccagaaaac
     1501 atccagtaaa ggagaggaac tatttacagg tactgtgcct atcaaagtcg agctggacgg
     1561 cgacgtaaac ggccacaagt tcagcgtgcg tggcgagggc gagggcgatg ccaccgaggg
     1621 caagctgacc ctgaagttca tctgcaccac cggcaagctg ccggtgccgt ggccgaccct
     1681 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc tacccggacc acatgaagcg
     1741 tcacgacttc ttcaagtccg ccatgccgga aggctacgtc caggagcgca ccatcgagtt
     1801 caaggacgac ggcacctaca agacccgcgc ggaggtgaag ttcgagggcg acaccctggt
     1861 gaaccgcatc gagctgaagg gcaacgactt caaggaggac ggcaacatcc tgggtcacaa
     1921 gctggagtac aaccacaaca gccacaacgt ccgtatcgaa gccgacaagc agaagaacgg
     1981 catcaaggct aacttcaaga tccgccacaa cgttgaggac ggcagccagc aggaagccga
     2041 ccacaaacag cagaacaccc cgatcggcga cggcccggtg cgcctgccgg acaaccacta
     2101 tctgagcacc cagaccacgc tgagcaaaga cccgaacgag aagcgcgatc acatggtcct
     2161 gaaggagttc gtgacggcag ccggtatcac taaaggtgaa gatgaaagtg gatccaatgc
     2221 cgaacctaaa cacaattcca atcggcagag cgaacgttct ggccggtccg gtgggaatcc
     2281 tggtggtagt ggtaatcaag gaggcagtgg aaattcacgc gggggtggtg ctggcagtgg
     2341 caataatcaa gggagtaaca gcggaggcgg gagtaattcg ggcgcttctt cgagcaatcc
     2401 agcgatgatg gcggctgccc aggcggccct tcaaagttcc tgggggatga tgggtatgct
     2461 tgcatcacaa cagaaccaaa gcgggcctag cggtaataat cagaaccagg gcaatagcca
     2521 gagagagcct aaccaggcta gtggcagtgg gaacaactcg tccagcggta gcaactcggg
     2581 agcagcctct ggttctggca gtgcttctaa cgctggtagc ggatcaggca gtaatggcgg
     2641 ctccggttct tcttctgatt ccaaatcttc cggttcaggt agttagaccc agctttcttg
     2701 tacaaagtgg ttgatcctct cgagcctcta gaactatagt gagtcgtatt acgtagatcc
     2761 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2821 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2881 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     2941 ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt
     3001 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     3061 tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa
     3121 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3181 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3241 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3301 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3361 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3421 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3481 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3541 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3601 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3661 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3721 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3781 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3841 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3901 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3961 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4021 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4081 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4141 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4201 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4261 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4321 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4381 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4441 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4501 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4561 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4621 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4681 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4741 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4801 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4861 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4921 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4981 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5041 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5101 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5161 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     5221 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     5281 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     5341 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     5401 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     5461 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     5521 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     5581 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     5641 agggcgcgtc ccattcgcca ttcaggctgc gcaactg
//
