LOCUS       sequence-471946-        8148 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SRSF1 mCherry fusion protein without MCP domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8148)
  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 8148)
  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-SRSF1-V5-mCherry
FEATURES             Location/Qualifiers
     source          1..8148
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             24..623
                     /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(24..43)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     520..540
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(704..721)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    710..934
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(981..997)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(981..997)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(994..1016)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1005..1021
                     /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(1029..1035,1036..1053,1054..1059))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1074..1095
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1154..1349
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1200..1335
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1355..1489
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1557..1574)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1728..2316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1808..1827)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2487..3278,3279..3347))
                     /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     3110..3129
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    3940..4074
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        4088..4467
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          4751..5767
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     5691..5713
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     5775..5794
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             5840..5878
                     /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             5888..5908
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5912..5935
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    5945..5969
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6714..6738)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6746..6787
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6788..7498
                     /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(6933..6951)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7201..7220
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    join(7585..8148,1..11,12..14,15..23)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7752..7769)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7979..7998
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
ORIGIN
        1 acgatgataa tatggccaca accatgaccg agtacaagcc cacggtgcgc ctcgccaccc
       61 gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
      121 cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct
      181 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
      241 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
      301 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
      361 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
      421 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
      481 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
      541 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
      601 tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga
      661 gcgcacgacc ccatgcatcg atgatctaga gctcgctgat cagcctcgac tgtgccttct
      721 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
      781 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
      841 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
      901 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aacctgcagc
      961 ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     1021 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     1081 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     1141 tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     1201 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     1261 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     1321 gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg
     1381 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     1441 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa
     1501 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     1561 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     1621 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     1681 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1741 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1801 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1861 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1921 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1981 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2041 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2101 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2161 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2221 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2281 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2341 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2401 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2461 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2521 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     2581 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     2641 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     2701 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     2761 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     2821 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     2881 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2941 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3001 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3061 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3121 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3181 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3241 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3301 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3361 caatgaccgc tttggaaaaa caaagactgt atttcctgga aattaatgtt tattcaataa
     3421 actgtgtatt cagctatatt cacatagtgg tgaggctgaa atgaggcggg aagaggcggt
     3481 tggggcttaa ttatatcaat ttgggtggcc ccacagcgcc tccaaggcgc cagtcctgtt
     3541 ttgacaagtt gcctctggaa gcctctcttc tttttctcca gagtaagcgg aggccagggg
     3601 cccccggcct ctgcttaata ctaaaaaaaa cagctgttgt catagtaatg attgggtgga
     3661 aacattccag gcctgggtgg agaggctttt tgcttcctct tgcaaaacca cactgacatt
     3721 ccaggcctgg gtggagaggc tttttgcttc ctcttgcaaa accacactgc cctctggagg
     3781 gcagttgcct agcaactaat taaaagagga tgtcgcacgg ccagctgcgg tcagttagtc
     3841 acttcctgct taactgactt gacattttct attttaagag tcgggaggaa aattactgtg
     3901 ttggaggccc tccgccatct tctgaagctg aatcgaatta acttgtttat tgcagcttat
     3961 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4021 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgatat
     4081 catcgtcgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     4141 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     4201 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     4261 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     4321 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     4381 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     4441 tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc
     4501 cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt
     4561 gcagcgatgg gggcgggggg gggggggggg ggggcgcgag gcgggggggg gggggcgggg
     4621 cggggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct
     4681 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
     4741 gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc
     4801 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
     4861 cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg
     4921 gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg
     4981 gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg
     5041 ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag
     5101 cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt
     5161 gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc
     5221 ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg
     5281 tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc
     5341 gggcggggcg gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg
     5401 gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg
     5461 cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg
     5521 ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat
     5581 gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg
     5641 gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt
     5701 ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt
     5761 cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt
     5821 cctcgaggct agcaccatgg cgcacatcgt gatggtggac gcctacaagc cgacgaaggg
     5881 cagcggtgag aacctgtact tccagagcgg cgactacaag gatgacgatg acaagggcgc
     5941 gccaacaagt ttgtacaaaa aagttggcat gtcgggaggt ggtgtgattc gtggccccgc
     6001 agggaacaac gattgccgca tctacgtggg taacttacct ccagacatcc gaaccaagga
     6061 cattgaggac gtgttctaca aatacggcgc tatccgcgac atcgacctca agaatcgccg
     6121 cgggggaccg cccttcgcct tcgttgagtt cgaggacccg cgagacgcgg aagacgcggt
     6181 gtatggtcgc gacggctatg attacgatgg gtaccgtctg cgggtggagt ttcctcgaag
     6241 cggccgtgga acaggccgag gcggcggcgg gggtggaggt ggcggagctc cccgaggtcg
     6301 ctatggcccc ccatccaggc ggtctgaaaa cagagtggtt gtctctggac tgcctccaag
     6361 tggaagttgg caggatttaa aggatcacat gcgtgaagca ggtgatgtat gttatgctga
     6421 tgtttaccga gatggcactg gtgtcgtgga gtttgtacgg aaagaagata tgacctatgc
     6481 agttcgaaaa ctggataaca ctaagtttag atctcatgag ggagaaactg cctacatccg
     6541 ggttaaagtt gatgggccca gaagtccaag ttatggaaga tctcgatctc gaagccgtag
     6601 tcgtagcaga agccgtagca gaagcaacag caggagtcgc agttactccc caaggagaag
     6661 cagaggatca ccacgctatt ctccccgtca tagcagatct cgctctcgta catacccaac
     6721 tttcttgtac aaagtggttg atatcggtaa gcctatccct aaccctctcc tcggtctcga
     6781 ttctacgatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
     6841 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     6901 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     6961 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     7021 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     7081 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     7141 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     7201 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     7261 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     7321 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     7381 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     7441 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtagta
     7501 atgaactagt accggttaag tcgacaatca acgcgttaag tcgacaggcc gctcgataag
     7561 cttgatatcg aattccgccc cccccccctc tccctccccc ccccctaacg ttactggccg
     7621 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     7681 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     7741 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     7801 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     7861 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     7921 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     7981 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     8041 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     8101 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaac
//
