LOCUS       sequence-203714-        6373 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flow cytometry-compatible reporter for TDP43 function in splicing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6373)
  TITLE     Rajat Rohatgi TDP43 plasmids
REFERENCE   2  (bases 1 to 6373)
  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: pHBS956 IBB-GFP-mCherry3E
FEATURES             Location/Qualifiers
     source          1..6373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    108..132
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      151..160
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1782..1800)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2050..2069
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(2353..2377)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2422..2556
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2446..2465)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2500..2519
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2669..2689)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2669..2687)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2708..2724)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2708..2724)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2721..2743)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2732..2748
                     /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(2756..2762,2763..2780,2781..2786))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2801..2822
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2939..2956)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3110..3698)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3190..3209)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3869..4660,4661..4729))
                     /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     4492..4511
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4730..4834)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4860..5315)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4997..5018)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5209..5228
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5389..6373
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattca aggatcaaca agtttgtaca
      121 aaaaagcagg ctggcgccgg taccaccggt gccaccatgg gatccaccaa cgagaatgct
      181 aatacaccag ctgcccgtct tcacagattc aagaacaagg gaaaagacag tacagaaatg
      241 aggcgtcgca gaatagaggt caatgtggag ctgaggaaag ctaagaagga tgaccagatg
      301 ctgaagagga gaaatgtaag ctcattccct gatgatggta ccagtaaagg agaggaacta
      361 tttacaggta ctgtgcctat caaagtcgag ctggacggcg acgtaaacgg ccacaagttc
      421 agcgtgcgtg gcgagggcga gggcgatgcc accgagggca agctgaccct gaagttcatc
      481 tgcaccaccg gcaagctgcc ggtgccgtgg ccgaccctcg tgaccaccct gacctacggc
      541 gtgcagtgct tcagccgcta cccggaccac atgaagcgtc acgacttctt caagtccgcc
      601 atgccggaag gctacgtcca ggagcgcacc atcgagttca aggacgacgg cacctacaag
      661 acccgcgcgg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
      721 aacgacttca aggaggacgg caacatcctg ggtcacaagc tggagtacaa ccacaacagc
      781 cacaacgtcc gtatcgaagc cgacaagcag aagaacggca tcaaggctaa cttcaagatc
      841 cgccacaacg ttgaggacgg cagccagcag gaagccgacc acaaacagca gaacaccccg
      901 atcggcgacg gcccggtgcg cctgccggac aaccactatc tgagcaccca gaccacgctg
      961 agcaaagacc cgaacgagaa gcgcgatcac atggtcctga aggagttcgt gacggcagcc
     1021 ggtatcacta aaggtgaaga tgaaagtgga ggcaccggaa ccagcaaggg cgaggaggat
     1081 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     1141 ggccacgagt tcgagatcga gggcgagggc gaggtgagta tggtacataa aacaagcatc
     1201 tattgaaaat atctgacaaa ctcatctttt atttttgatg tgtgtgtgtg tgtgtgtgtg
     1261 tgtgtttttt tttaacaggg atttggggaa taatttgaga aagcaaaaca aaacaataac
     1321 aatagaaaaa cttctaatgg tgatgacagc ctcttcttca gtaatttctc acttcttggt
     1381 actcctgtcc tgaaagatat taatttcaag atagaaagag gacagttgtt ggcggttgct
     1441 ggatccactg gagcaggcaa ggtagttctt ttgttcttca ctattaagaa cttaatttgg
     1501 tgtccatgtc tctttttttt tctagtttgt agtgctggaa ggtatttttg gagaaattct
     1561 tacatgagca ttaggagaat gtatgggtgt agtgtcttgt ataatagaaa ttgttccact
     1621 gataatttac tctagttttt tatttcctca tattattttc agtggctttt tcttccacat
     1681 ctttatattt tgcaccacat tcaacactgt atcttgcaca tggcgagcat gccctttcct
     1741 tttttttcct cagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     1801 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
     1861 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
     1921 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     1981 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     2041 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
     2101 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     2161 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     2221 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     2281 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     2341 caagtaaact agcttcagct ttcttgtaca aagtggttga tcctctcgag cctctagaac
     2401 tatagtgagt cgtattacgt agatccagac atgataagat acattgatga gtttggacaa
     2461 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     2521 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     2581 atgtttcagg ttcaggggga ggtgtgggag gttttttaat tcgcggccgc ggcgccaatg
     2641 cattgggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     2701 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     2761 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     2821 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     2881 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     2941 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3001 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3061 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3121 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3181 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3241 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3301 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3361 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3421 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3481 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3541 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3601 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3661 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3721 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3781 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3841 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3901 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3961 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4021 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4081 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4141 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4201 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4261 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4321 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4381 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4441 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4501 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4561 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4621 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4681 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4741 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4801 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     4861 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     4921 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     4981 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     5041 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     5101 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     5161 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     5221 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     5281 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     5341 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagtcga ccatagccaa
     5401 ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa
     5461 ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa
     5521 ttcaatatgg cggacttggc accatgccaa ttcaatatgg cggacttggc actgtgccaa
     5581 ctggggaggg gtctacttgg cacggtgcca agtttgagga ggggtcttgg ccctgtgcca
     5641 agtccgccat attgaattgg catggtgcca ataatggcgg ccatattggc tatatgccag
     5701 gatcaatata taggcaatat ccaatatggc cctatgccaa tatggctatt ggccaggttc
     5761 aatactatgt attggcccta tgccatatag tattccatat atgggttttc ctattgacgt
     5821 agatagcccc tcccaatggg cggtcccata taccatatat ggggcttcct aataccgccc
     5881 atagccactc ccccattgac gtcaatggtc tctatatatg gtctttccta ttgacgtcat
     5941 atgggcggtc ctattgacgt atatggcgcc tcccccattg acgtcaatta cggtaaatgg
     6001 cccgcctggc tcaatgccca ttgacgtcaa taggaccacc caccattgac gtcaatggga
     6061 tggctcattg cccattcata tccgttctca cgccccctat tgacgtcaat gacggtaaat
     6121 ggcccacttg gcagtacatc aatatctatt aatagtaact tggcaagtac attactattg
     6181 gaaggacgcc agggtacatt ggcagtactc ccattgacgt caatggcggt aaatggcccg
     6241 cgatggctgc caagtacatc cccattgacg tcaatgggga ggggcaatga cgcaaatggg
     6301 cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata taagcaatgc
     6361 tcgtttaggg aac
//
