LOCUS       sequence-471949-        7890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SRSF3 mCherry fusion protein without MCP domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7890)
  AUTHORS   Bhat P, Chow A, Emert B, Ettlin O, Quinodoz SA, Strehle M, Takei Y, 
            Burr A, Goronzy IN, Chen AW, Huang W, Ferrer JLM, Soehalim E, Goh 
            ST, Chari T, Sullivan DK, Blanco MR, Guttman M
  TITLE     Genome organization around nuclear speckles drives mRNA splicing 
            efficiency.
  JOURNAL   Nature. 2024 May;629(8014):1165-1173. doi: 
            10.1038/s41586-024-07429-6. Epub 2024 May 8.
  PUBMED    38720076
REFERENCE   2  (bases 1 to 7890)
  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"; doi: 
            "10.1038/s41586-024-07429-6"; journalName: "Nature"; date: 
            "2024-05"; volume: "629"; issue: "8014"; pages: "1165-1173"
COMMENT     Sequence Label: pCAG-SpyTag-Flag-SRSF3-V5-mCherry
FEATURES             Location/Qualifiers
     source          1..7890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1624..1758
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        1772..2151
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          2429..3445
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3369..3391
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3453..3472
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3518..3556
                     /codon_start=1
                     /product="peptide derived from the CnaB2 adhesin domain of 
                     Streptococcus pyogenes FbaB"
                     /label=SpyTag
                     /note="forms a covalent isopeptide bond with the SpyCatcher
                     protein (Zakeri et al., 2012)"
                     /translation="AHIVMVDAYKPTK"
     CDS             3566..3586
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3590..3613
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    3623..3647
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4140..4164)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4172..4213
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4214..4924
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4359..4377)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4627..4646
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    join(5011..5585,5586..5588,5589..5597)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5178..5195)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5405..5424
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5598..6197
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5598..5617)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6094..6114
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6278..6295)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6284..6508
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6555..6571)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6555..6571)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6568..6590)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6579..6595
                     /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(6603..6609,6610..6627,6628..6633))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6648..6669
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6728..6923
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6774..6909
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6929..7063
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7131..7148)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7302..7890)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7382..7401)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatga ccgctttgga aaaacaaaga ctgtatttcc
     1081 tggaaattaa tgtttattca ataaactgtg tattcagcta tattcacata gtggtgaggc
     1141 tgaaatgagg cgggaagagg cggttggggc ttaattatat caatttgggt ggccccacag
     1201 cgcctccaag gcgccagtcc tgttttgaca agttgcctct ggaagcctct cttctttttc
     1261 tccagagtaa gcggaggcca ggggcccccg gcctctgctt aatactaaaa aaaacagctg
     1321 ttgtcatagt aatgattggg tggaaacatt ccaggcctgg gtggagaggc tttttgcttc
     1381 ctcttgcaaa accacactga cattccaggc ctgggtggag aggctttttg cttcctcttg
     1441 caaaaccaca ctgccctctg gagggcagtt gcctagcaac taattaaaag aggatgtcgc
     1501 acggccagct gcggtcagtt agtcacttcc tgcttaactg acttgacatt ttctatttta
     1561 agagtcggga ggaaaattac tgtgttggag gccctccgcc atcttctgaa gctgaatcga
     1621 attaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     1681 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     1741 tcttatcatg tctggatctg atatcatcgt cgacattgat tattgactag ttattaatag
     1801 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     1861 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     1921 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     1981 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     2041 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     2101 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg
     2161 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat
     2221 ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggsgcgcgcc
     2281 rggggggggg gggggggggg gggggggggg gggggsgrgg sggrgaggkg cggcggcagc
     2341 caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc
     2401 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc
     2461 ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca
     2521 ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg
     2581 gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg
     2641 cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg
     2701 ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg
     2761 cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga
     2821 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc
     2881 gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc
     2941 ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg
     3001 gcggcaggkg ggggtgccgg gcggggcggg gccgcctcgg gccggggrgg gctcgggggr
     3061 ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
     3121 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga
     3181 gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg
     3241 cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct
     3301 tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg
     3361 cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt
     3421 tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca
     3481 tcattttggc aaagaattcc tcgaggctag caccatggcg cacatcgtga tggtggacgc
     3541 ctacaagccg acgaagggca gcggtgagaa cctgtacttc cagagcggcg actacaagga
     3601 tgacgatgac aagggcgcgc caacaagttt gtacaaaaaa gttggcatgc atcgtgattc
     3661 ctgtccattg gactgtaagg tttatgtagg caatcttgga aacaatggca acaagacgga
     3721 attggaacgg gcttttggct actatggacc actccgaagt gtgtgggttg ctagaaaccc
     3781 acccggcttt gcttttgttg aatttgaaga tccccgagat gcagctgatg cagtccgaga
     3841 gctagatgga agaacactat gtggctgccg tgtaagagtg gaactgtcga atggtgaaaa
     3901 aagaagtaga aatcgtggcc cacctccctc ttggggtcgt cgccctcgag atgattatcg
     3961 taggaggagt cctccacctc gtcgcagatc tccaagaagg agaagcttct ctcgcagccg
     4021 gagcaggtcc ctttctagag ataggagaag agagagatcg ctgtctcggg agagaaatca
     4081 caagccgtcc cgatccttct ctaggtctcg tagtcgatct aggtcaaatg aaaggaaata
     4141 cccaactttc ttgtacaaag tggttgatat cggtaagcct atccctaacc ctctcctcgg
     4201 tctcgattct acgatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     4261 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     4321 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     4381 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     4441 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     4501 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     4561 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     4621 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     4681 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     4741 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     4801 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     4861 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     4921 gtagtaatga actagtaccg gttaagtcga caatcaacgc gttaagtcga caggccgctc
     4981 gataagcttg atatcgaatt ccgccccccc cccctctccc tccccccccc ctaacgttac
     5041 tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat
     5101 attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat
     5161 tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga
     5221 agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca
     5281 gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac
     5341 acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt
     5401 caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca
     5461 ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt
     5521 aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg
     5581 ataatatggc cacaaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     5641 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     5701 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     5761 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     5821 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     5881 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     5941 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     6001 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     6061 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     6121 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     6181 gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac
     6241 gaccccatgc atcgatgatc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     6301 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     6361 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     6421 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     6481 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaacctg cagcccaagc
     6541 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6601 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6661 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6721 cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     6781 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     6841 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     6901 ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata atggttacaa
     6961 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     7021 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca ttaatgaatc
     7081 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     7141 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     7201 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     7261 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     7321 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     7381 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     7441 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     7501 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     7561 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     7621 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7681 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7741 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7801 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7861 cagattacgc gcagaaaaaa aggatctcaa
//
