LOCUS       sequence-208207-        6374 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 6374)
  TITLE     Rajat Rohatgi TDP43 plasmids
REFERENCE   2  (bases 1 to 6374)
  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..6374
                     /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"
     primer_bind     complement(1781..1799)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2049..2068
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2425..2559
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2449..2468)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2503..2522
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2672..2692)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2672..2690)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2711..2727)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2711..2727)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2724..2746)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2735..2751
                     /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(2759..2765,2766..2783,2784..2789))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2804..2825
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2942..2959)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3113..3701)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3193..3212)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3872..4663,4664..4732))
                     /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     4495..4514
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4733..4837)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4863..5318)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5000..5021)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5212..5231
                     /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 gccattctgc ctggggacgt cggagcaagc ttgatttagg tgacactata gaatacaagc
       61 tacttgttct ttttgcagga tcccatcgat tcgaattcaa ggatcaacaa gtttgtacaa
      121 aaaagcaggc tggcgccggt accaccggtg ccaccatggg atccaccaac gagaatgcta
      181 atacaccagc tgcccgtctt cacagattca agaacaaggg aaaagacagt acagaaatga
      241 ggcgtcgcag aatagaggtc aatgtggagc tgaggaaagc taagaaggat gaccagatgc
      301 tgaagaggag aaatgtaagc tcattccctg atgatggtac cagtaaagga gaggaactat
      361 ttacaggtac tgtgcctatc aaagtcgagc tggacggcga cgtaaacggc cacaagttca
      421 gcgtgcgtgg cgagggcgag ggcgatgcca ccgagggcaa gctgaccctg aagttcatct
      481 gcaccaccgg caagctgccg gtgccgtggc cgaccctcgt gaccaccctg acctacggcg
      541 tgcagtgctt cagccgctac ccggaccaca tgaagcgtca cgacttcttc aagtccgcca
      601 tgccggaagg ctacgtccag gagcgcacca tcgagttcaa ggacgacggc acctacaaga
      661 cccgcgcgga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
      721 acgacttcaa ggaggacggc aacatcctgg gtcacaagct ggagtacaac cacaacagcc
      781 acaacgtccg tatcgaagcc gacaagcaga agaacggcat caaggctaac ttcaagatcc
      841 gccacaacgt tgaggacggc agccagcagg aagccgacca caaacagcag aacaccccga
      901 tcggcgacgg cccggtgcgc ctgccggaca accactatct gagcacccag accacgctga
      961 gcaaagaccc gaacgagaag cgcgatcaca tggtcctgaa ggagttcgtg acggcagccg
     1021 gtatcactaa aggtgaagat gaaagtggag gcaccggaac cagcaagggc gaggaggata
     1081 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     1141 gccacgagtt cgagatcgag ggcgagggcg aggtgagtat ggtacataaa acaagcatct
     1201 attgaaaata tctgacaaac tcatctttta tttttgatgt gtgtgtgtgt gtgtgtgtgt
     1261 gtgttttttt ttaacaggga tttggggaat aatttgagaa agcaaaacaa aacaataaca
     1321 atagaaaaac ttctaatggt gatgacagcc tcttcttcag taatttctca cttcttggta
     1381 ctcctgtcct gaaagatatt aatttcaaga tagaaagagg acagttgttg gcggttgctg
     1441 gatccactgg agcaggcaag gtagttcttt tgttcttcac tattaagaac ttaatttggt
     1501 gtccatgtct cttttttttt ctagtttgta gtgctggaag gtatttttgg agaaattctt
     1561 acatgagcat taggagaatg tatgggtgta gtgtcttgta taatagaaat tgttccactg
     1621 ataatttact ctagtttttt atttcctcat attattttca gtggcttttt cttccacatc
     1681 tttatatttt gcaccacatt caacactgta tcttgcacat ggcgagcatg ccctttcctt
     1741 ttttttcctc agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     1801 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
     1861 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc
     1921 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
     1981 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc
     2041 tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg
     2101 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc
     2161 ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     2221 ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc
     2281 gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac
     2341 aagtaagcta tctagaccca gctttcttgt acaaagtgat tgatcctctc gagcctctag
     2401 aactatagtg agtcgtatta cgtagatcca gacatgataa gatacattga tgagtttgga
     2461 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2521 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     2581 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aattcgcggc cgcggcgcca
     2641 atgcattggg cccggtaccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     2701 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     2761 acatacgagc cgggagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     2821 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     2881 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2941 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3001 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3061 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3121 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3181 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3241 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3301 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3361 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3421 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3481 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3541 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3601 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3661 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3721 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3781 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3841 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3901 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3961 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4021 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4081 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4141 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4201 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4261 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4321 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4381 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4441 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4501 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4561 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4621 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4681 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4741 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4801 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4861 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     4921 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     4981 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     5041 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     5101 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     5161 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     5221 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     5281 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
     5341 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt cgaccgccaa
     5401 ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa
     5461 ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa
     5521 ttcaatatgg cggacttggc accatgccaa ttcaatatgg cggacctggc 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 gaagtacgcc agggtacatt ggcagtactc ccattgacgt caatggcggt aaatggcccg
     6241 cgatggctgc caagtacatc cccattgacg tcaatgggga ggggcaatga cgcaaatggg
     6301 cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata taagcaatgc
     6361 tcgtttaggg aacc
//
