LOCUS       sequence-470053-        7815 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SRRM1 lacking Nuclear Speckle domain (dNS-SRRM1) mCherry fusion 
            protein without MCP domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7815)
  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 7815)
  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-dNS-SRRM1-V5-mCherry
FEATURES             Location/Qualifiers
     source          1..7815
                     /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(4065..4089)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4097..4138
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4139..4849
                     /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(4284..4302)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4552..4571
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    join(4936..5510,5511..5513,5514..5522)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5103..5120)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5330..5349
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5523..6122
                     /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(5523..5542)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6019..6039
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6203..6220)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6209..6433
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6480..6496)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6480..6496)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6493..6515)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6504..6520
                     /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(6528..6534,6535..6552,6553..6558))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6573..6594
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6653..6848
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6699..6834
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6854..6988
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7056..7073)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7227..7815)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7307..7326)
                     /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 ggggcgcgcc
     2281 rggggggggg gggggggggg gggggggggg sggggsgrgg 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 gccgcckcgg 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 gcaggctcca ccatggacgc
     3661 gggatttttc cgcggaacaa gtgcagaaca ggataatcgg ttcagcaaca aacagaagaa
     3721 actactgaag cagctgaaat ttgcagaatg cctagaaaaa aaggtggaca tgagcaaagt
     3781 aaatttggag gttataaagc cttggataac aaaaagagta acggaaatcc ttgggtttga
     3841 agatgatgtt gtgattgagt ttatattcaa ccagctggaa gtgaagaatc cagactccaa
     3901 aatgatgcaa atcaacctga ctggattttt gaatggaaaa aatgctcgag aatttatggg
     3961 agaactgtgg cccctgctgc taagtgcaca agaaaacatc gcgggaatcc cttctgcttt
     4021 cctagaactg aagaaagaag aaataaaaca aagacagatt gaacaaccag ctttcttgta
     4081 caaagtggtt gatatcggta agcctatccc taaccctctc ctcggtctcg attctacgat
     4141 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     4201 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     4261 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     4321 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     4381 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     4441 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     4501 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     4561 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     4621 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     4681 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     4741 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     4801 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtagt aatgaactag
     4861 taccggttaa gtcgacaatc aacgcgttaa gtcgacaggc cgctcgataa gcttgatatc
     4921 gaattccgcc ccccccccct ctccctcccc cccccctaac gttactggcc gaagccgctt
     4981 ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg
     5041 caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc
     5101 ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga
     5161 agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc
     5221 tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc
     5281 acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc
     5341 aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga
     5401 tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg
     5461 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa
     5521 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     5581 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     5641 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     5701 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     5761 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     5821 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     5881 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     5941 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     6001 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     6061 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     6121 gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcga
     6181 tgatctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     6241 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     6301 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     6361 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     6421 gtgggctcta tggcttctga ggcggaaaga acctgcagcc caagcttggc gtaatcatgg
     6481 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6541 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6601 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
     6661 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     6721 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6781 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     6841 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6901 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6961 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
     7021 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7081 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7141 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7201 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7261 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7321 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7381 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7441 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7501 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7561 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7621 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     7681 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7741 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7801 aaaaaaggat ctcaa
//
