LOCUS       sequence-229668-        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: pHBS1383 TDP43 RRM-GFP FYW-S
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        42..1019
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1054..1072
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1054..1071
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1127..1151
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1170..1179
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2679..2703)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2748..2882
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2772..2791)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2826..2845
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2995..3015)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2995..3013)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3034..3050)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3034..3050)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3047..3069)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3058..3074
                     /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(3082..3088,3089..3106,3107..3112))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3127..3148
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3265..3282)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3436..4024)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3516..3535)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4195..4986,4987..5055))
                     /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     4818..4837
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5056..5160)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5186..5641)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5323..5344)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5535..5554
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggcgatcggt gcgggcctct tcgctattac gccagtcgac cgccaattca atatggcgta
       61 tatggactca tgccaattca atatggtgga tctggacctg tgccaattca atatggcgta
      121 tatggactcg tgccaattca atatggtgga tctggacccc agccaattca atatggcgga
      181 cttggcacca tgccaattca atatggcgga cctggcactg tgccaactgg ggaggggtct
      241 acttggcacg gtgccaagtt tgaggagggg tcttggccct gtgccaagtc cgccatattg
      301 aattggcatg gtgccaataa tggcggccat attggctata tgccaggatc aatatatagg
      361 caatatccaa tatggcccta tgccaatatg gctattggcc aggttcaata ctatgtattg
      421 gccctatgcc atatagtatt ccatatatgg gttttcctat tgacgtagat agcccctccc
      481 aatgggcggt cccatatacc atatatgggg cttcctaata ccgcccatag ccactccccc
      541 attgacgtca atggtctcta tatatggtct ttcctattga cgtcatatgg gcggtcctat
      601 tgacgtatat ggcgcctccc ccattgacgt caattacggt aaatggcccg cctggctcaa
      661 tgcccattga cgtcaatagg accacccacc attgacgtca atgggatggc tcattgccca
      721 ttcatatccg ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc cacttggcag
      781 tacatcaata tctattaata gtaacttggc aagtacatta ctattggaag tacgccaggg
      841 tacattggca gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag
      901 tacatcccca ttgacgtcaa tggggagggg caatgacgca aatgggcgtt ccattgacgt
      961 aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt ttagggaacc
     1021 gccattctgc ctggggacgt cggagcaagc ttgatttagg tgacactata gaatacaagc
     1081 tacttgttct ttttgcagga tcccatcgat tcgaattcaa ggatcaacaa gtttgtacaa
     1141 aaaagcaggc tggcgccggt accaccggtg ccaccatggg atccatgtct gaatatattc
     1201 gggtaaccga agatgagaac gatgagccca ttgaaatacc atcggaagac gatgggacgg
     1261 tgctgctctc cacggttaca gcccagtttc caggggcgtg tgggcttcgc tacaggaatc
     1321 cagtgtctca gtgtatgaga ggtgtccggc tggtagaagg aattctgcat gccccagatg
     1381 ctggctgggg aaatctggtg tatgttgtca actatccaaa agataacaaa agaaaaatgg
     1441 atgagacaga tgcttcatca gcagtgaaag tgaaaagagc agtccagaaa acatccagta
     1501 aaggagagga actatttaca ggtactgtgc ctatcaaagt cgagctggac ggcgacgtaa
     1561 acggccacaa gttcagcgtg cgtggcgagg gcgagggcga tgccaccgag ggcaagctga
     1621 ccctgaagtt catctgcacc accggcaagc tgccggtgcc gtggccgacc ctcgtgacca
     1681 ccctgaccta cggcgtgcag tgcttcagcc gctacccgga ccacatgaag cgtcacgact
     1741 tcttcaagtc cgccatgccg gaaggctacg tccaggagcg caccatcgag ttcaaggacg
     1801 acggcaccta caagacccgc gcggaggtga agttcgaggg cgacaccctg gtgaaccgca
     1861 tcgagctgaa gggcaacgac ttcaaggagg acggcaacat cctgggtcac aagctggagt
     1921 acaaccacaa cagccacaac gtccgtatcg aagccgacaa gcagaagaac ggcatcaagg
     1981 ctaacttcaa gatccgccac aacgttgagg acggcagcca gcaggaagcc gaccacaaac
     2041 agcagaacac cccgatcggc gacggcccgg tgcgcctgcc ggacaaccac tatctgagca
     2101 cccagaccac gctgagcaaa gacccgaacg agaagcgcga tcacatggtc ctgaaggagt
     2161 tcgtgacggc agccggtatc actaaaggtg aagatgaaag tggatccaat gccgaaccta
     2221 agcataatag caacagacaa ctcgaacgaa gcgggcggag tgggggcaac cccggcggat
     2281 ctggtaatca aggcggaagc ggcaatagcc ggggtggggg agccggtctc ggcaataatc
     2341 aaggatctaa catgggcggc ggtatgaact ctggcgcatc cagtatcaac ccggccatga
     2401 tggcagcagc acaggccgcc ctgcaaagtt catggggaat gatgggtatg ttggcaagtc
     2461 agcagaacca gtcaggacct tccgggaata atcaaaacca agggaacatg caacgcgaac
     2521 ccaatcaagc ctcaggatca ggcaataata gcagttccgg ctcaaattcc ggcgcggcga
     2581 tagggtctgg ctccgcctca aatgcgggaa gcggatcagg gagtaacgga ggatcaggct
     2641 catctatgga ttcaaaatcc agcggcagtg gtatgtagac ccagctttct tgtacaaagt
     2701 ggttgatcct ctcgagcctc tagaactata gtgagtcgta ttacgtagat ccagacatga
     2761 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     2821 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2881 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     2941 tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta
     3001 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3061 ttatccgctc acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg
     3121 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3181 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3241 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3301 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3361 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3421 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3481 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3541 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3601 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3661 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3721 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3781 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3841 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3901 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3961 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4021 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4081 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4141 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4201 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4261 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4321 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4381 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4441 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4501 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4561 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4621 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4681 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4741 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4801 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4861 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4921 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4981 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5041 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5101 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5161 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     5221 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     5281 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     5341 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     5401 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     5461 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     5521 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     5581 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     5641 tcccattcgc cattcaggct gcgcaactgt tgggaag
//
