LOCUS       sequence-207933-        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: pHBS961 TDP43 RRM-GFP G295S
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..41
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     23..40
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    96..120
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      139..148
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1648..1672)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1717..1851
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1741..1760)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1795..1814
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1964..1984)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1964..1982)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2003..2019)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2003..2019)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2016..2038)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2027..2043
                     /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(2051..2057,2058..2075,2076..2081))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2096..2117
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2234..2251)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2405..2993)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2485..2504)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3164..3955,3956..4024))
                     /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     3787..3806
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4025..4129)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4155..4610)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4292..4313)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4504..4523
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4681..5665
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 tggggacgtc ggagcaagct tgatttaggt gacactatag aatacaagct acttgttctt
       61 tttgcaggat cccatcgatt cgaattcaag gatcaacaag tttgtacaaa aaagcaggct
      121 ggcgccggta ccaccggtgc caccatggga tccatgtctg aatatattcg ggtaaccgaa
      181 gatgagaacg atgagcccat tgaaatacca tcggaagacg atgggacggt gctgctctcc
      241 acggttacag cccagtttcc aggggcgtgt gggcttcgct acaggaatcc agtgtctcag
      301 tgtatgagag gtgtccggct ggtagaagga attctgcatg ccccagatgc tggctgggga
      361 aatctggtgt atgttgtcaa ctatccaaaa gataacaaaa gaaaaatgga tgagacagat
      421 gcttcatcag cagtgaaagt gaaaagagca gtccagaaaa catccagtaa aggagaggaa
      481 ctatttacag gtactgtgcc tatcaaagtc gagctggacg gcgacgtaaa cggccacaag
      541 ttcagcgtgc gtggcgaggg cgagggcgat gccaccgagg gcaagctgac cctgaagttc
      601 atctgcacca ccggcaagct gccggtgccg tggccgaccc tcgtgaccac cctgacctac
      661 ggcgtgcagt gcttcagccg ctacccggac cacatgaagc gtcacgactt cttcaagtcc
      721 gccatgccgg aaggctacgt ccaggagcgc accatcgagt tcaaggacga cggcacctac
      781 aagacccgcg cggaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
      841 ggcaacgact tcaaggagga cggcaacatc ctgggtcaca agctggagta caaccacaac
      901 agccacaacg tccgtatcga agccgacaag cagaagaacg gcatcaaggc taacttcaag
      961 atccgccaca acgttgagga cggcagccag caggaagccg accacaaaca gcagaacacc
     1021 ccgatcggcg acggcccggt gcgcctgccg gacaaccact atctgagcac ccagaccacg
     1081 ctgagcaaag acccgaacga gaagcgcgat cacatggtcc tgaaggagtt cgtgacggca
     1141 gccggtatca ctaaaggtga agatgaaagt ggatccaatg ccgaacctaa gcacaatagc
     1201 aatagacagt tagaaagaag tggaagattt ggtggtaatc caggtggctt tgggaatcag
     1261 ggtggatttg gtaatagcag agggtctgga gctggtttgg gaaacaatca aggtagtaat
     1321 atgggtggtg ggatgaactt tggtgcgttc agcattaatc cagccatgat ggctgccgcc
     1381 caggcagcac tacagagcag ttggggtatg atgggcatgt tagccagcca gcagaaccag
     1441 tcaggcccat cgggtaataa ccaaaaccaa ggcaacatgc agagggagcc aaaccaggcc
     1501 ttcggttctg gaaataactc ttatagtggc tctaattctg gtgcagcaat tggttgggga
     1561 tcagcatcca atgcagggtc gggcagtggt tttaatggag gctttggctc aagcatggat
     1621 tctaagtctt ctggctgggg aatgtagacc cagctttctt gtacaaagtg gttgatcctc
     1681 tcgagcctct agaactatag tgagtcgtat tacgtagatc cagacatgat aagatacatt
     1741 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     1801 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     1861 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg
     1921 gccgcggcgc caatgcattg ggcccggtac ccagcttttg ttccctttag tgagggttaa
     1981 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     2041 caattccaca caacatacga gccgggagca taaagtgtaa agcctggggt gcctaatgag
     2101 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     2161 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2221 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2281 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2341 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2401 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2461 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2521 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2581 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2641 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2701 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2761 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2821 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2881 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2941 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3001 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3061 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3121 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3181 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3241 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3301 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3361 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3421 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3481 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3541 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3601 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3661 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3721 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3781 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3841 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     3901 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     3961 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4021 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4081 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4141 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     4201 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     4261 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     4321 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     4381 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     4441 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     4501 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     4561 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
     4621 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     4681 gtcgaccgcc aattcaatat ggcgtatatg gactcatgcc aattcaatat ggtggatctg
     4741 gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc aattcaatat ggtggatctg
     4801 gaccccagcc aattcaatat ggcggacttg gcaccatgcc aattcaatat ggcggacctg
     4861 gcactgtgcc aactggggag gggtctactt ggcacggtgc caagtttgag gaggggtctt
     4921 ggccctgtgc caagtccgcc atattgaatt ggcatggtgc caataatggc ggccatattg
     4981 gctatatgcc aggatcaata tataggcaat atccaatatg gccctatgcc aatatggcta
     5041 ttggccaggt tcaatactat gtattggccc tatgccatat agtattccat atatgggttt
     5101 tcctattgac gtagatagcc cctcccaatg ggcggtccca tataccatat atggggcttc
     5161 ctaataccgc ccatagccac tcccccattg acgtcaatgg tctctatata tggtctttcc
     5221 tattgacgtc atatgggcgg tcctattgac gtatatggcg cctcccccat tgacgtcaat
     5281 tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc aataggacca cccaccattg
     5341 acgtcaatgg gatggctcat tgcccattca tatccgttct cacgccccct attgacgtca
     5401 atgacggtaa atggcccact tggcagtaca tcaatatcta ttaatagtaa cttggcaagt
     5461 acattactat tggaagtacg ccagggtaca ttggcagtac tcccattgac gtcaatggcg
     5521 gtaaatggcc cgcgatggct gccaagtaca tccccattga cgtcaatggg gaggggcaat
     5581 gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg cgtgcctaat gggaggtcta
     5641 tataagcaat gctcgtttag ggaaccgcca ttctgcc
//
