LOCUS       sequence-207927-        5677 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 5677)
  TITLE     Rajat Rohatgi TDP43 plasmids
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"
COMMENT     Sequence Label: pHBS1276 TDP43 RRM-GFP W385D
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        31..49
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     31..48
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    104..128
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      147..156
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1656..1680)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1725..1859
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1749..1768)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1803..1822
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1972..1992)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1972..1990)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2011..2027)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2011..2027)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2024..2046)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2035..2051
                     /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(2059..2065,2066..2083,2084..2089))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2104..2125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2242..2259)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2413..3001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2493..2512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3172..3963,3964..4032))
                     /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     3795..3814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4033..4137)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4163..4618)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4300..4321)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4512..4531
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4689..5673
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 attctgcctg gggacgtcgg agcaagcttg atttaggtga cactatagaa tacaagctac
       61 ttgttctttt tgcaggatcc catcgattcg aattcaagga tcaacaagtt tgtacaaaaa
      121 agcaggctgg cgccggtacc accggtgcca ccatgggatc catgtctgaa tatattcggg
      181 taaccgaaga tgagaacgat gagcccattg aaataccatc ggaagacgat gggacggtgc
      241 tgctctccac ggttacagcc cagtttccag gggcgtgtgg gcttcgctac aggaatccag
      301 tgtctcagtg tatgagaggt gtccggctgg tagaaggaat tctgcatgcc ccagatgctg
      361 gctggggaaa tctggtgtat gttgtcaact atccaaaaga taacaaaaga aaaatggatg
      421 agacagatgc ttcatcagca gtgaaagtga aaagagcagt ccagaaaaca tccagtaaag
      481 gagaggaact atttacaggt actgtgccta tcaaagtcga gctggacggc gacgtaaacg
      541 gccacaagtt cagcgtgcgt ggcgagggcg agggcgatgc caccgagggc aagctgaccc
      601 tgaagttcat ctgcaccacc ggcaagctgc cggtgccgtg gccgaccctc gtgaccaccc
      661 tgacctacgg cgtgcagtgc ttcagccgct acccggacca catgaagcgt cacgacttct
      721 tcaagtccgc catgccggaa ggctacgtcc aggagcgcac catcgagttc aaggacgacg
      781 gcacctacaa gacccgcgcg gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
      841 agctgaaggg caacgacttc aaggaggacg gcaacatcct gggtcacaag ctggagtaca
      901 accacaacag ccacaacgtc cgtatcgaag ccgacaagca gaagaacggc atcaaggcta
      961 acttcaagat ccgccacaac gttgaggacg gcagccagca ggaagccgac cacaaacagc
     1021 agaacacccc gatcggcgac ggcccggtgc gcctgccgga caaccactat ctgagcaccc
     1081 agaccacgct gagcaaagac ccgaacgaga agcgcgatca catggtcctg aaggagttcg
     1141 tgacggcagc cggtatcact aaaggtgaag atgaaagtgg atccaatgcc gaacctaagc
     1201 acaatagcaa tagacagtta gaaagaagtg gaagatttgg tggtaatcca ggtggctttg
     1261 ggaatcaggg tggatttggt aatagcagag ggggtggagc tggtttggga aacaatcaag
     1321 gtagtaatat gggtggtggg atgaactttg gtgcgttcag cattaatcca gccatgatgg
     1381 ctgccgccca ggcagcacta cagagcagtt ggggtatgat gggcatgtta gccagccagc
     1441 agaaccagtc aggcccatcg ggtaataacc aaaaccaagg caacatgcag agggagccaa
     1501 accaggcctt cggttctgga aataactctt atagtggctc taattctggt gcagcaattg
     1561 gtgatggatc agcatccaat gcagggtcgg gcagtggttt taatggaggc tttggctcaa
     1621 gcatggattc taagtcttct ggctggggaa tgtagaccca gctttcttgt acaaagtggt
     1681 tgatcctctc gagcctctag aactatagtg agtcgtatta cgtagatcca gacatgataa
     1741 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     1801 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     1861 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     1921 aattcgcggc cgcggcgcca atgcattggg cccggtaccc agcttttgtt ccctttagtg
     1981 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     2041 tccgctcaca attccacaca acatacgagc cgggagcata aagtgtaaag cctggggtgc
     2101 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2161 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2221 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2281 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2341 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2401 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2461 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2521 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2581 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2641 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2701 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2761 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2821 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2881 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2941 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3001 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3061 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3121 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3181 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3241 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3301 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3361 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3421 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3481 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3541 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3601 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3661 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3721 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3781 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3841 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3901 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3961 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4021 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4081 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4141 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
     4201 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     4261 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     4321 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     4381 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     4441 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     4501 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     4561 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     4621 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     4681 ttacgccagt cgaccgccaa ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg
     4741 tggatctgga cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg
     4801 tggatctgga ccccagccaa ttcaatatgg cggacttggc accatgccaa ttcaatatgg
     4861 cggacctggc actgtgccaa ctggggaggg gtctacttgg cacggtgcca agtttgagga
     4921 ggggtcttgg ccctgtgcca agtccgccat attgaattgg catggtgcca ataatggcgg
     4981 ccatattggc tatatgccag gatcaatata taggcaatat ccaatatggc cctatgccaa
     5041 tatggctatt ggccaggttc aatactatgt attggcccta tgccatatag tattccatat
     5101 atgggttttc ctattgacgt agatagcccc tcccaatggg cggtcccata taccatatat
     5161 ggggcttcct aataccgccc atagccactc ccccattgac gtcaatggtc tctatatatg
     5221 gtctttccta ttgacgtcat atgggcggtc ctattgacgt atatggcgcc tcccccattg
     5281 acgtcaatta cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa taggaccacc
     5341 caccattgac gtcaatggga tggctcattg cccattcata tccgttctca cgccccctat
     5401 tgacgtcaat gacggtaaat ggcccacttg gcagtacatc aatatctatt aatagtaact
     5461 tggcaagtac attactattg gaagtacgcc agggtacatt ggcagtactc ccattgacgt
     5521 caatggcggt aaatggcccg cgatggctgc caagtacatc cccattgacg tcaatgggga
     5581 ggggcaatga cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg
     5641 gaggtctata taagcaatgc tcgtttaggg aaccgcc
//
