LOCUS       sequence-338317-        5252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of FLAG-tagged, codon-optimized, truncated SRSF3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5252)
  AUTHORS   Campbell AE, Dyle MC, Albanese R, Matheny T, Sudheendran K, Cortazar
            MA, Forman T, Fu R, Gillen AE, Caruthers MH, Floor SN, Calviello L, 
            Jagannathan S
  TITLE     Compromised nonsense-mediated RNA decay results in truncated 
            RNA-binding protein production upon DUX4 expression.
  JOURNAL   Cell Rep. 2023 Jun 13;42(6):112642. doi: 
            10.1016/j.celrep.2023.112642.
  PUBMED    37314931
REFERENCE   2  (bases 1 to 5252)
  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"; journalName: "Cell Rep.";
            date: "2023-06-13"; volume: "42(6):112642. doi"; pages: " 
            10.1016/j.celrep.2023.112642"
COMMENT     Sequence Label: pTwist-FLAG-SRSF3_Truncated_Codon.Optimized
FEATURES             Location/Qualifiers
     source          1..5252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      170..306
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    458..529
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(534..580)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    598..646
                     /note="synthetic polyadenylation signal"
     promoter        683..1861
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          914..1852
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1809..1829
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1961..1984
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2425..2546
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2436..2455)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2490..2509
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2661..2707
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2737..2873
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2918..2989
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3215..3803)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3295..3314)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4549..4617,4618..5252,1..157)
                     /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     complement(4767..4786)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
       61 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
      121 cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg cattggcgca
      181 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
      241 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
      301 ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga
      361 gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc
      421 gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca
      481 ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca attcaaaaaa
      541 cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac
      601 acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat
      661 cgagtgtagc cagatctcgt gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc
      721 cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg
      781 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
      841 gggagaaccg tatataagtg cactagtcgc cgtgaacgtt ctttttcgca acgggtttgc
      901 cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta
      961 tggcccttgc gtgccttgaa ttacttccac ctggctgcag tacgtgattc ttgatcccga
     1021 gcttcgggtt ggaagtgggt gggagagttc gtggccttgc gcttaaggag ccccttcgcc
     1081 tcgtgcttga gttgtggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca
     1141 ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc
     1201 tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag atcagcacac
     1261 tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg
     1321 ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc
     1381 tgcccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc
     1441 aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc
     1501 tgcagggagc acaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac
     1561 acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg
     1621 ggcgccgtcc aggcacctcg attagttctc cagcttttgg agtacgtcgt ctttaggttg
     1681 gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag
     1741 gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg
     1801 gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg
     1861 aaaactaccc ctaaaagcca aaagatcttt gtcgatccta ccatccactc gacacacccg
     1921 ccagcggccg ctgccaagct tccgagctct cgaattcatg gactacaaag atgacgacga
     1981 taaaatgcac cgagactcct gcccgctgga ttgcaaggta tatgtaggga atctcggcaa
     2041 taatggcaac aagacggaac ttgaaagagc atttggctat tatggccctc tgcggagtgt
     2101 atgggttgcc agaaacccgc caggtttcgc gttcgtcgaa tttgaagacc cacgagacgc
     2161 cgctgacgcg gttagggagc tggacgggcg cacactctgt ggctgcaggg tgagggtgga
     2221 actcagcaat ggcgagaaga ggtcccgaaa tcgcggccct ccaccatcct gggggcgccg
     2281 accaagggac gattaccgac gccgctctcc tccccctcga agacgagtca ctataatgag
     2341 ccttcttaca acgctgtgag gatccgcagg cctctgctag cttgactgac tgagatacag
     2401 cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa accacaacta
     2461 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     2521 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     2581 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg
     2641 gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg
     2701 ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct
     2761 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt
     2821 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc
     2881 agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg
     2941 gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt
     3001 taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt
     3061 gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt
     3121 gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg
     3181 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3241 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3301 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3361 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3421 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3481 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3541 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3601 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3661 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3721 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3781 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3841 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3901 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3961 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4021 ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc
     4081 ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct
     4141 cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca
     4201 atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg
     4261 atctatccga gatccgagga atatcgaaat cggggcgcgc ctggtgtacc gagaacgatc
     4321 ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca
     4381 gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt
     4441 accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta
     4501 gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa
     4561 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4621 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac
     4681 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt
     4741 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4801 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca
     4861 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4921 ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag
     4981 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5041 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5101 gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag
     5161 ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5221 gctggctggt ttattgctga taaatctgga gc
//
