LOCUS       sequence-235269-        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: pHBS1387 TDP43 RRM-GFP KRED-S
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        52..1036
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1071..1089
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1071..1088
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1144..1168
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1187..1196
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2696..2720)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2765..2899
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2789..2808)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2843..2862
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3012..3032)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3012..3030)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3051..3067)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3051..3067)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3064..3086)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3075..3091
                     /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(3099..3105,3106..3123,3124..3129))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3144..3165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3282..3299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3453..4041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3533..3552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4212..5003,5004..5072))
                     /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     4835..4854
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5073..5177)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5203..5658)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5340..5361)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5552..5571
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agtcgaccgc
       61 caattcaata tggcgtatat ggactcatgc caattcaata tggtggatct ggacctgtgc
      121 caattcaata tggcgtatat ggactcgtgc caattcaata tggtggatct ggaccccagc
      181 caattcaata tggcggactt ggcaccatgc caattcaata tggcggacct ggcactgtgc
      241 caactgggga ggggtctact tggcacggtg ccaagtttga ggaggggtct tggccctgtg
      301 ccaagtccgc catattgaat tggcatggtg ccaataatgg cggccatatt ggctatatgc
      361 caggatcaat atataggcaa tatccaatat ggccctatgc caatatggct attggccagg
      421 ttcaatacta tgtattggcc ctatgccata tagtattcca tatatgggtt ttcctattga
      481 cgtagatagc ccctcccaat gggcggtccc atataccata tatggggctt cctaataccg
      541 cccatagcca ctcccccatt gacgtcaatg gtctctatat atggtctttc ctattgacgt
      601 catatgggcg gtcctattga cgtatatggc gcctccccca ttgacgtcaa ttacggtaaa
      661 tggcccgcct ggctcaatgc ccattgacgt caataggacc acccaccatt gacgtcaatg
      721 ggatggctca ttgcccattc atatccgttc tcacgccccc tattgacgtc aatgacggta
      781 aatggcccac ttggcagtac atcaatatct attaatagta acttggcaag tacattacta
      841 ttggaagtac gccagggtac attggcagta ctcccattga cgtcaatggc ggtaaatggc
      901 ccgcgatggc tgccaagtac atccccattg acgtcaatgg ggaggggcaa tgacgcaaat
      961 gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa tgggaggtct atataagcaa
     1021 tgctcgttta gggaaccgcc attctgcctg gggacgtcgg agcaagcttg atttaggtga
     1081 cactatagaa tacaagctac ttgttctttt tgcaggatcc catcgattcg aattcaagga
     1141 tcaacaagtt tgtacaaaaa agcaggctgg cgccggtacc accggtgcca ccatgggatc
     1201 catgtctgaa tatattcggg taaccgaaga tgagaacgat gagcccattg aaataccatc
     1261 ggaagacgat gggacggtgc tgctctccac ggttacagcc cagtttccag gggcgtgtgg
     1321 gcttcgctac aggaatccag tgtctcagtg tatgagaggt gtccggctgg tagaaggaat
     1381 tctgcatgcc ccagatgctg gctggggaaa tctggtgtat gttgtcaact atccaaaaga
     1441 taacaaaaga aaaatggatg agacagatgc ttcatcagca gtgaaagtga aaagagcagt
     1501 ccagaaaaca tccagtaaag gagaggaact atttacaggt actgtgccta tcaaagtcga
     1561 gctggacggc gacgtaaacg gccacaagtt cagcgtgcgt ggcgagggcg agggcgatgc
     1621 caccgagggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc cggtgccgtg
     1681 gccgaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct acccggacca
     1741 catgaagcgt cacgacttct tcaagtccgc catgccggaa ggctacgtcc aggagcgcac
     1801 catcgagttc aaggacgacg gcacctacaa gacccgcgcg gaggtgaagt tcgagggcga
     1861 caccctggtg aaccgcatcg agctgaaggg caacgacttc aaggaggacg gcaacatcct
     1921 gggtcacaag ctggagtaca accacaacag ccacaacgtc cgtatcgaag ccgacaagca
     1981 gaagaacggc atcaaggcta acttcaagat ccgccacaac gttgaggacg gcagccagca
     2041 ggaagccgac cacaaacagc agaacacccc gatcggcgac ggcccggtgc gcctgccgga
     2101 caaccactat ctgagcaccc agaccacgct gagcaaagac ccgaacgaga agcgcgatca
     2161 catggtcctg aaggagttcg tgacggcagc cggtatcact aaaggtgaag atgaaagtgg
     2221 atccaatgcc gaacctagcc ataactctaa ttctcaactc agttccagtg gatctttcgg
     2281 cggaaatcct ggaggttttg gaaaccaagg tggttttggg aatagttctg gaggaggtgc
     2341 tggcctgggt aataatcaag ggagtaatat gggaggtgga atgaattttg gtgctttctc
     2401 aataaatccc gctatgatgg cggctgcgca agcagccttg caaagttcct gggggatgat
     2461 gggcatgttg gcttctcagc agaaccagag cgggccgtcc ggcaataatc aaaaccaagg
     2521 gaatatgcag tcaagcccta accaagcatt cggcagcgga aataattcct atagtggttc
     2581 aaattctggt gcggccattg ggtggggctc cgcttccaat gctggcagtg gaagtggatt
     2641 caatgggggg ttcggctctt caatgtcatc ttctagttct ggttggggaa tgtagaccca
     2701 gctttcttgt acaaagtggt tgatcctctc gagcctctag aactatagtg agtcgtatta
     2761 cgtagatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     2821 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     2881 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     2941 ggaggtgtgg gaggtttttt aattcgcggc cgcggcgcca atgcattggg cccggtaccc
     3001 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     3061 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cgggagcata
     3121 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3181 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3241 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3301 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3361 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3421 aggaaccgta aaaaggccgc gttgctggcc tttttccata ggctccgccc ccctgacgag
     3481 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3541 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3601 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3661 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3721 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3781 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3841 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3901 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3961 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4021 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4081 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4141 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4201 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4261 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4321 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4381 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4441 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4501 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4561 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4621 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4681 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4741 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4801 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4861 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4921 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4981 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5041 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5101 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5161 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     5221 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     5281 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     5341 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     5401 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     5461 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     5521 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     5581 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     5641 cgccgctaca gggcgcgtcc cattcgccat tcaggct
//
