LOCUS       sequence-207964-        5682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To test the effect of sequence on TDP43 droplet properties.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5682)
  TITLE     Rajat Rohatgi TDP43 plasmids
REFERENCE   2  (bases 1 to 5682)
  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"
COMMENT     Sequence Label: pHBS975 TDP43 RRM-GFP N345D
FEATURES             Location/Qualifiers
     source          1..5682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..45
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     27..44
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    100..124
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      143..152
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1657..1681)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1726..1860
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1750..1769)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1804..1823
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1973..1993)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1973..1991)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2012..2028)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2012..2028)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2025..2047)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2036..2052
                     /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(2060..2066,2067..2084,2085..2090))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2105..2126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2243..2260)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2414..3002)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2494..2513)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3173..3964,3965..4033))
                     /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     3796..3815
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4034..4138)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4164..4619)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4301..4322)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4513..4532
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4690..5674
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 tgcctgggga cgtcggagca agcttgattt aggtgacact atagaataca agctacttgt
       61 tctttttgca ggatcccatc gattcgaatt caaggatcaa caagtttgta caaaaaagca
      121 ggctggcgcc ggtaccaccg gtgccaccat gggatccatg tctgaatata ttcgggtaac
      181 cgaagatgag aacgatgagc ccattgaaat accatcggaa gacgatggga cggtgctgct
      241 ctccacggtt acagcccagt ttccaggggc gtgtgggctt cgctacagga atccagtgtc
      301 tcagtgtatg agaggtgtcc ggctggtaga aggaattctg catgccccag atgctggctg
      361 gggaaatctg gtgtatgttg tcaactatcc aaaagataac aaaagaaaaa tggatgagac
      421 agatgcttca tcagcagtga aagtgaaaag agcagtccag aaaacatcca gtaaaggaga
      481 ggaactattt acaggtactg tgcctatcaa agtcgagctg gacggcgacg taaacggcca
      541 caagttcagc gtgcgtggcg agggcgaggg cgatgccacc gagggcaagc tgaccctgaa
      601 gttcatctgc accaccggca agctgccggt gccgtggccg accctcgtga ccaccctgac
      661 ctacggcgtg cagtgcttca gccgctaccc ggaccacatg aagcgtcacg acttcttcaa
      721 gtccgccatg ccggaaggct acgtccagga gcgcaccatc gagttcaagg acgacggcac
      781 ctacaagacc cgcgcggagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
      841 gaagggcaac gacttcaagg aggacggcaa catcctgggt cacaagctgg agtacaacca
      901 caacagccac aacgtccgta tcgaagccga caagcagaag aacggcatca aggctaactt
      961 caagatccgc cacaacgttg aggacggcag ccagcaggaa gccgaccaca aacagcagaa
     1021 caccccgatc ggcgacggcc cggtgcgcct gccggacaac cactatctga gcacccagac
     1081 cacgctgagc aaagacccga acgagaagcg cgatcacatg gtcctgaagg agttcgtgac
     1141 ggcagccggt atcactaaag gtgaagatga aagtggatcc aatgccgaac ctaagcacaa
     1201 tagcaataga cagttagaaa gaagtggaag atttggtggt aatccaggtg gctttgggaa
     1261 tcagggtgga tttggtaata gcagaggggg tggagctggt ttgggaaaca atcaaggtag
     1321 taatatgggt ggtgggatga actttggtgc gttcagcatt aatccagcca tgatggctgc
     1381 cgcccaggca gcactacaga gcagttgggg tatgatgggc atgttagcca gccagcagga
     1441 ccagtcaggc ccatcgggta ataaccaaaa ccaaggcaac atgcagaggg agccaaacca
     1501 ggccttcggt tctggaaata actcttatag tggctctaat tctggtgcag caattggttg
     1561 gggatcagca tccaatgcag ggtcgggcag tggttttaat ggaggctttg gctcaagcat
     1621 ggattctaag tcttctggct ggggaatgta gactagaccc agctttcttg tacaaagtgg
     1681 ttgatcctct cgagcctcta gaactatagt gagtcgtatt acgtagatcc agacatgata
     1741 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     1801 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     1861 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     1921 taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt tccctttagt
     1981 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2041 atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa gcctggggtg
     2101 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2161 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2221 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2281 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2341 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2401 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2461 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2521 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2581 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2641 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2701 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2761 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2821 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2881 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2941 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3001 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3061 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3121 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3181 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3241 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3301 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3361 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3421 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3481 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3541 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3601 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3661 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3721 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3781 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3841 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3901 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3961 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4021 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4081 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4141 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
     4201 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     4261 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     4321 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     4381 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     4441 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     4501 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     4561 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     4621 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     4681 attacgccag tcgaccgcca attcaatatg gcgtatatgg actcatgcca attcaatatg
     4741 gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg
     4801 gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg
     4861 gcggacctgg cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg
     4921 aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg
     4981 gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca
     5041 atatggctat tggccaggtt caatactatg tattggccct atgccatata gtattccata
     5101 tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata
     5161 tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt ctctatatat
     5221 ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt
     5281 gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac
     5341 ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta
     5401 ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac
     5461 ttggcaagta cattactatt ggaagtacgc cagggtacat tggcagtact cccattgacg
     5521 tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg
     5581 aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg
     5641 ggaggtctat ataagcaatg ctcgtttagg gaaccgccat tc
//
