LOCUS       sequence-210060-        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: pHBS959 TDP43 RRM-GFP G295F
FEATURES             Location/Qualifiers
     source          1..5682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        34..52
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     34..51
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    107..131
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      150..159
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1664..1688)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1733..1867
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1757..1776)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1811..1830
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1980..2000)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1980..1998)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2019..2035)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2019..2035)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2032..2054)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2043..2059
                     /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(2067..2073,2074..2091,2092..2097))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2112..2133
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2250..2267)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2421..3009)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2501..2520)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3180..3971,3972..4040))
                     /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     3803..3822
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4041..4145)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4171..4626)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4308..4329)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4520..4539
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4697..5681
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 gccattctgc ctggggacgt cggagcaagc ttgatttagg tgacactata gaatacaagc
       61 tacttgttct ttttgcagga tcccatcgat tcgaattcaa ggatcaacaa gtttgtacaa
      121 aaaagcaggc tggcgccggt accaccggtg ccaccatggg atccatgtct gaatatattc
      181 gggtaaccga agatgagaac gatgagccca ttgaaatacc atcggaagac gatgggacgg
      241 tgctgctctc cacggttaca gcccagtttc caggggcgtg tgggcttcgc tacaggaatc
      301 cagtgtctca gtgtatgaga ggtgtccggc tggtagaagg aattctgcat gccccagatg
      361 ctggctgggg aaatctggtg tatgttgtca actatccaaa agataacaaa agaaaaatgg
      421 atgagacaga tgcttcatca gcagtgaaag tgaaaagagc agtccagaaa acatccagta
      481 aaggagagga actatttaca ggtactgtgc ctatcaaagt cgagctggac ggcgacgtaa
      541 acggccacaa gttcagcgtg cgtggcgagg gcgagggcga tgccaccgag ggcaagctga
      601 ccctgaagtt catctgcacc accggcaagc tgccggtgcc gtggccgacc ctcgtgacca
      661 ccctgaccta cggcgtgcag tgcttcagcc gctacccgga ccacatgaag cgtcacgact
      721 tcttcaagtc cgccatgccg gaaggctacg tccaggagcg caccatcgag ttcaaggacg
      781 acggcaccta caagacccgc gcggaggtga agttcgaggg cgacaccctg gtgaaccgca
      841 tcgagctgaa gggcaacgac ttcaaggagg acggcaacat cctgggtcac aagctggagt
      901 acaaccacaa cagccacaac gtccgtatcg aagccgacaa gcagaagaac ggcatcaagg
      961 ctaacttcaa gatccgccac aacgttgagg acggcagcca gcaggaagcc gaccacaaac
     1021 agcagaacac cccgatcggc gacggcccgg tgcgcctgcc ggacaaccac tatctgagca
     1081 cccagaccac gctgagcaaa gacccgaacg agaagcgcga tcacatggtc ctgaaggagt
     1141 tcgtgacggc agccggtatc actaaaggtg aagatgaaag tggatccaat gccgaaccta
     1201 agcacaatag caatagacag ttagaaagaa gtggaagatt tggtggtaat ccaggtggct
     1261 ttgggaatca gggtggattt ggtaatagca gagggtttgg agctggtttg ggaaacaatc
     1321 aaggtagtaa tatgggtggt gggatgaact ttggtgcgtt cagcattaat ccagccatga
     1381 tggctgccgc ccaggcagca ctacagagca gttggggtat gatgggcatg ttagccagcc
     1441 agcagaacca gtcaggccca tcgggtaata accaaaacca aggcaacatg cagagggagc
     1501 caaaccaggc cttcggttct ggaaataact cttatagtgg ctctaattct ggtgcagcaa
     1561 ttggttgggg atcagcatcc aatgcagggt cgggcagtgg ttttaatgga ggctttggct
     1621 caagcatgga ttctaagtct tctggctggg gaatgtagac tagacccagc tttcttgtac
     1681 aaagtggttg atcctctcga gcctctagaa ctatagtgag tcgtattacg tagatccaga
     1741 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     1801 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     1861 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     1921 ggttttttaa ttcgcggccg cggcgccaat gcattgggcc cggtacccag cttttgttcc
     1981 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     2041 aattgttatc cgctcacaat tccacacaac atacgagccg ggagcataaa gtgtaaagcc
     2101 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2161 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2221 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2281 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2341 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2401 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2461 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2521 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2581 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2641 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2701 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2761 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2821 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2881 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2941 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3001 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3061 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3121 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3181 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3241 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3301 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3361 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3421 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3481 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3541 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3601 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3661 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3721 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3781 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3841 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3901 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     3961 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4021 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4081 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4141 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     4201 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     4261 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     4321 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     4381 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     4441 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     4501 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4561 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4621 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     4681 cttcgctatt acgccagtcg accgccaatt caatatggcg tatatggact catgccaatt
     4741 caatatggtg gatctggacc tgtgccaatt caatatggcg tatatggact cgtgccaatt
     4801 caatatggtg gatctggacc ccagccaatt caatatggcg gacttggcac catgccaatt
     4861 caatatggcg gacctggcac tgtgccaact ggggaggggt ctacttggca cggtgccaag
     4921 tttgaggagg ggtcttggcc ctgtgccaag tccgccatat tgaattggca tggtgccaat
     4981 aatggcggcc atattggcta tatgccagga tcaatatata ggcaatatcc aatatggccc
     5041 tatgccaata tggctattgg ccaggttcaa tactatgtat tggccctatg ccatatagta
     5101 ttccatatat gggttttcct attgacgtag atagcccctc ccaatgggcg gtcccatata
     5161 ccatatatgg ggcttcctaa taccgcccat agccactccc ccattgacgt caatggtctc
     5221 tatatatggt ctttcctatt gacgtcatat gggcggtcct attgacgtat atggcgcctc
     5281 ccccattgac gtcaattacg gtaaatggcc cgcctggctc aatgcccatt gacgtcaata
     5341 ggaccaccca ccattgacgt caatgggatg gctcattgcc cattcatatc cgttctcacg
     5401 ccccctattg acgtcaatga cggtaaatgg cccacttggc agtacatcaa tatctattaa
     5461 tagtaacttg gcaagtacat tactattgga agtacgccag ggtacattgg cagtactccc
     5521 attgacgtca atggcggtaa atggcccgcg atggctgcca agtacatccc cattgacgtc
     5581 aatggggagg ggcaatgacg caaatgggcg ttccattgac gtaaatgggc ggtaggcgtg
     5641 cctaatggga ggtctatata agcaatgctc gtttagggaa cc
//
