LOCUS       sequence-207950-        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: pHBS1251 TDP43 RRM-GFP G309C
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        40..58
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     40..57
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    113..137
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      156..165
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1665..1689)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1734..1868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1758..1777)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1812..1831
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1981..2001)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1981..1999)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2020..2036)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2020..2036)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2033..2055)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2044..2060
                     /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(2068..2074,2075..2092,2093..2098))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2113..2134
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2251..2268)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2422..3010)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2502..2521)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3181..3972,3973..4041))
                     /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     3804..3823
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4042..4146)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4172..4627)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4309..4330)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4521..4540
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(4698..5677,1..5)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 ggaaccgcca ttctgcctgg ggacgtcgga gcaagcttga tttaggtgac actatagaat
       61 acaagctact tgttcttttt gcaggatccc atcgattcga attcaaggat caacaagttt
      121 gtacaaaaaa gcaggctggc gccggtacca ccggtgccac catgggatcc atgtctgaat
      181 atattcgggt aaccgaagat gagaacgatg agcccattga aataccatcg gaagacgatg
      241 ggacggtgct gctctccacg gttacagccc agtttccagg ggcgtgtggg cttcgctaca
      301 ggaatccagt gtctcagtgt atgagaggtg tccggctggt agaaggaatt ctgcatgccc
      361 cagatgctgg ctggggaaat ctggtgtatg ttgtcaacta tccaaaagat aacaaaagaa
      421 aaatggatga gacagatgct tcatcagcag tgaaagtgaa aagagcagtc cagaaaacat
      481 ccagtaaagg agaggaacta tttacaggta ctgtgcctat caaagtcgag ctggacggcg
      541 acgtaaacgg ccacaagttc agcgtgcgtg gcgagggcga gggcgatgcc accgagggca
      601 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc ggtgccgtgg ccgaccctcg
      661 tgaccaccct gacctacggc gtgcagtgct tcagccgcta cccggaccac atgaagcgtc
      721 acgacttctt caagtccgcc atgccggaag gctacgtcca ggagcgcacc atcgagttca
      781 aggacgacgg cacctacaag acccgcgcgg aggtgaagtt cgagggcgac accctggtga
      841 accgcatcga gctgaagggc aacgacttca aggaggacgg caacatcctg ggtcacaagc
      901 tggagtacaa ccacaacagc cacaacgtcc gtatcgaagc cgacaagcag aagaacggca
      961 tcaaggctaa cttcaagatc cgccacaacg ttgaggacgg cagccagcag gaagccgacc
     1021 acaaacagca gaacaccccg atcggcgacg gcccggtgcg cctgccggac aaccactatc
     1081 tgagcaccca gaccacgctg agcaaagacc cgaacgagaa gcgcgatcac atggtcctga
     1141 aggagttcgt gacggcagcc ggtatcacta aaggtgaaga tgaaagtgga tccaatgccg
     1201 aacctaagca caatagcaat agacagttag aaagaagtgg aagatttggt ggtaatccag
     1261 gtggctttgg gaatcagggt ggatttggta atagcagagg gggtggagct ggtttgggaa
     1321 acaatcaagg tagtaatatg ggttgtggga tgaactttgg tgcgttcagc attaatccag
     1381 ccatgatggc tgccgcccag gcagcactac agagcagttg gggtatgatg ggcatgttag
     1441 ccagccagca gaaccagtca ggcccatcgg gtaataacca aaaccaaggc aacatgcaga
     1501 gggagccaaa ccaggccttc ggttctggaa ataactctta tagtggctct aattctggtg
     1561 cagcaattgg ttggggatca gcatccaatg cagggtcggg cagtggtttt aatggaggct
     1621 ttggctcaag catggattct aagtcttctg gctggggaat gtagacccag ctttcttgta
     1681 caaagtggtt gatcctctcg agcctctaga actatagtga gtcgtattac gtagatccag
     1741 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     1801 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     1861 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     1921 aggtttttta attcgcggcc gcggcgccaa tgcattgggc ccggtaccca gcttttgttc
     1981 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     2041 aaattgttat ccgctcacaa ttccacacaa catacgagcc gggagcataa agtgtaaagc
     2101 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     2161 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     2221 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2281 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2341 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2401 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2461 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2521 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2581 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2641 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2701 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2761 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2821 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2881 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2941 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3001 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3061 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3121 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3181 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3241 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3301 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3361 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3421 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3481 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3541 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3601 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3661 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3721 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3781 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3841 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3901 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3961 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4021 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4081 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4141 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     4201 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     4261 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     4321 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     4381 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     4441 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     4501 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     4561 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     4621 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     4681 tcttcgctat tacgccagtc gaccgccaat tcaatatggc gtatatggac tcatgccaat
     4741 tcaatatggt ggatctggac ctgtgccaat tcaatatggc gtatatggac tcgtgccaat
     4801 tcaatatggt ggatctggac cccagccaat tcaatatggc ggacttggca ccatgccaat
     4861 tcaatatggc ggacctggca ctgtgccaac tggggagggg tctacttggc acggtgccaa
     4921 gtttgaggag gggtcttggc cctgtgccaa gtccgccata ttgaattggc atggtgccaa
     4981 taatggcggc catattggct atatgccagg atcaatatat aggcaatatc caatatggcc
     5041 ctatgccaat atggctattg gccaggttca atactatgta ttggccctat gccatatagt
     5101 attccatata tgggttttcc tattgacgta gatagcccct cccaatgggc ggtcccatat
     5161 accatatatg gggcttccta ataccgccca tagccactcc cccattgacg tcaatggtct
     5221 ctatatatgg tctttcctat tgacgtcata tgggcggtcc tattgacgta tatggcgcct
     5281 cccccattga cgtcaattac ggtaaatggc ccgcctggct caatgcccat tgacgtcaat
     5341 aggaccaccc accattgacg tcaatgggat ggctcattgc ccattcatat ccgttctcac
     5401 gccccctatt gacgtcaatg acggtaaatg gcccacttgg cagtacatca atatctatta
     5461 atagtaactt ggcaagtaca ttactattgg aagtacgcca gggtacattg gcagtactcc
     5521 cattgacgtc aatggcggta aatggcccgc gatggctgcc aagtacatcc ccattgacgt
     5581 caatggggag gggcaatgac gcaaatgggc gttccattga cgtaaatggg cggtaggcgt
     5641 gcctaatggg aggtctatat aagcaatgct cgtttag
//
