LOCUS       sequence-229720-        6374 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Fluorescence-based TDP43-dependent splicing reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6374)
  AUTHORS   Schmidt HB, Barreau A, Rohatgi R
  TITLE     Phase separation-deficient TDP43 remains functional in splicing.
  JOURNAL   Nat Commun. 2019 Oct 25;10(1):4890. doi: 10.1038/s41467-019-12740-2.
  PUBMED    31653829
REFERENCE   2  (bases 1 to 6374)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2019-10-25"; volume: "10(1):4890. doi"; pages: " 
            10.1038/s41467-019-12740-2"
COMMENT     Sequence Label: pHBS1389 IBB-GFP-mCherry3E
FEATURES             Location/Qualifiers
     source          1..6374
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        49..1033
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1068..1086
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1068..1085
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1141..1165
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1184..1193
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2815..2833)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3083..3102
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3459..3593
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3483..3502)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3537..3556
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3706..3726)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3706..3724)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3745..3761)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3745..3761)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3758..3780)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3769..3785
                     /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(3793..3799,3800..3817,3818..3823))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3838..3859
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3976..3993)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4147..4735)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4227..4246)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4906..5697,5698..5766))
                     /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     5529..5548
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5767..5871)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5897..6352)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6034..6055)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6246..6265
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt cgaccgccaa
       61 ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa
      121 ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa
      181 ttcaatatgg cggacttggc accatgccaa ttcaatatgg cggacctggc actgtgccaa
      241 ctggggaggg gtctacttgg cacggtgcca agtttgagga ggggtcttgg ccctgtgcca
      301 agtccgccat attgaattgg catggtgcca ataatggcgg ccatattggc tatatgccag
      361 gatcaatata taggcaatat ccaatatggc cctatgccaa tatggctatt ggccaggttc
      421 aatactatgt attggcccta tgccatatag tattccatat atgggttttc ctattgacgt
      481 agatagcccc tcccaatggg cggtcccata taccatatat ggggcttcct aataccgccc
      541 atagccactc ccccattgac gtcaatggtc tctatatatg gtctttccta ttgacgtcat
      601 atgggcggtc ctattgacgt atatggcgcc tcccccattg acgtcaatta cggtaaatgg
      661 cccgcctggc tcaatgccca ttgacgtcaa taggaccacc caccattgac gtcaatggga
      721 tggctcattg cccattcata tccgttctca cgccccctat tgacgtcaat gacggtaaat
      781 ggcccacttg gcagtacatc aatatctatt aatagtaact tggcaagtac attactattg
      841 gaagtacgcc agggtacatt ggcagtactc ccattgacgt caatggcggt aaatggcccg
      901 cgatggctgc caagtacatc cccattgacg tcaatgggga ggggcaatga cgcaaatggg
      961 cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata taagcaatgc
     1021 tcgtttaggg aaccgccatt ctgcctgggg acgtcggagc aagcttgatt taggtgacac
     1081 tatagaatac aagctacttg ttctttttgc aggatcccat cgattcgaat tcaaggatca
     1141 acaagtttgt acaaaaaagc aggctggcgc aggtaccacc ggtgccacca tgggatccac
     1201 caacgagaat gctaatacac cagctgcccg tcttcacaga ttcaagaaca agggaaaaga
     1261 cagtacagaa atgaggcgtc gcagaataga ggtcaatgtg gagctgagga aagctaagaa
     1321 ggatgaccag atgctgaaga ggagaaatgt aagctcattc cctgatgatg gtaccagtaa
     1381 aggagaggaa ctatttacag gtactgtgcc tatcaaagtc gagctggacg gcgacgtaaa
     1441 cggccacaag ttcagcgtgc gtggcgaggg cgagggcgat gccaccgagg gcaagctgac
     1501 cctgaagttc atctgcacca ccggcaagct gccggtgccg tggccgaccc tcgtgaccac
     1561 cctgacctac ggcgtgcagt gcttcagccg ctacccggac cacatgaagc gtcacgactt
     1621 cttcaagtcc gccatgccgg aaggctacgt ccaggagcgc accatcgagt tcaaggacga
     1681 cggcacctac aagacccgcg cggaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1741 cgagctgaag ggcaacgact tcaaggagga cggcaacatc ctgggtcaca agctggagta
     1801 caaccacaac agccacaacg tccgtatcga agccgacaag cagaagaacg gcatcaaggc
     1861 taacttcaag atccgccaca acgttgagga cggcagccag caggaagccg accacaaaca
     1921 gcagaacacc ccgatcggcg acggcccggt gcgcctgccg gacaaccact atctgagcac
     1981 ccagaccacg ctgagcaaag acccgaacga gaagcgcgat cacatggtcc tgaaggagtt
     2041 cgtgacggca gccggtatca ctaaaggtga agatgaaagt ggaggcaccg gaaccagcaa
     2101 gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga
     2161 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgaggtga gtatggtaca
     2221 taaaacaagc atctattgaa aatatctgac aaactcatct tttatttttg atgtgtgtgt
     2281 gtgtgtgtgt gtgtgtgttt ttttttaaca gggatttggg gaattatttg agaaagcaaa
     2341 acaaaacaat aacaatagaa aaacttctaa tggtgatgac agcctcttct tcagtaattt
     2401 ctcacttctt ggtactcctg tcctgaaaga tattaatttc aagatagaaa gaggacagtt
     2461 gttggcggtt gctggatcca ctggagcagg caaggtagtt cttttgttct tcactattaa
     2521 gaacttaatt tggtgtccat gtctcttttt ttttctagtt tgtagtgctg gaaggtattt
     2581 ttggagaaat tcttacatga gcattaggag aatgtatggg tgtagtgtct tgtataatag
     2641 aaattgttcc actgataatt tactctagtt ttttatttcc tcatattatt ttcagtggct
     2701 ttttcttcca catctttata ttttgcacca cattcaacac tgtatcttgc acatggcgag
     2761 catgcccttt cctttttttt cctcagggcc gcccctacga gggcacccag accgccaagc
     2821 tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca
     2881 tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt
     2941 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga
     3001 ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg
     3061 gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct
     3121 cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga
     3181 agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc
     3241 ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg
     3301 aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca
     3361 tggacgagct gtacaagtaa gctatctaga cccagctttc ttgtacaaag tgattgatcc
     3421 tctcgagcct ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca
     3481 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     3541 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     3601 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg
     3661 cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt
     3721 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3781 cacaattcca cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg
     3841 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3901 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3961 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4021 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4081 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4141 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4201 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4261 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4321 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4381 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4441 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4501 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4561 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4621 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4681 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4741 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4801 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4861 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4921 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4981 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5041 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5101 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5161 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5221 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5281 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5341 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5401 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5461 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5521 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5581 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5641 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5701 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5761 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5821 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5881 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     5941 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     6001 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     6061 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     6121 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     6181 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     6241 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     6301 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     6361 ccattcaggc tgcg
//
