LOCUS       sequence-422663-        8115 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of SRSF2 mutant in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8115)
  AUTHORS   Kim E, Ilagan JO, Liang Y, Daubner GM, Lee SC, Ramakrishnan A, Li Y,
            Chung YR, Micol JB, Murphy ME, Cho H, Kim MK, Zebari AS, Aumann S, 
            Park CY, Buonamici S, Smith PG, Deeg HJ, Lobry C, Aifantis I, Modis 
            Y, Allain FH, Halene S, Bradley RK, Abdel-Wahab O
  TITLE     SRSF2 Mutations Contribute to Myelodysplasia by Mutant-Specific 
            Effects on Exon Recognition.
  JOURNAL   Cancer Cell. 2015 May 11;27(5):617-30. doi: 
            10.1016/j.ccell.2015.04.006.
  PUBMED    25965569
REFERENCE   2  (bases 1 to 8115)
  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: 
            "10.1016/j.ccell.2015.04"; journalName: "Cancer Cell"; date: 
            "2015-05-11- 11"; volume: "27"; issue: "5"; pages: "617-30"
COMMENT     Sequence Label: pRRL_SRSF2_P95H_mCherry
FEATURES             Location/Qualifiers
     source          1..8115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..234
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             235..415
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    462..587
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1080..1313
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1498..1542
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1809..1926
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1972..2482
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2401..2419
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3163..3186
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3187..3240
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3241..3951
                     /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(3386..3404)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3654..3673
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3966..4554
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4019..4039)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4437..4448)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4641..4874
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4946..5067
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4983..5002)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5037..5056
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5107..5242
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5262..5281)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5263..5281)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5291..5308)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5291..5307)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5300..5322)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5449..5904
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5536..5555)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5746..5767
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5930..6034
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6035..6103,6104..6895)
                     /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(6253..6272)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7066..7654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7555..7574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7808..7825
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(7978..7983,7984..8001,8002..8008)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8016..8032
                     /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)."
     primer_bind     8021..8043
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8040..8056
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8040..8056
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8075..8095
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8077..8095
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
       61 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
      121 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
      181 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
      241 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      301 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      361 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
      421 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
      481 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
      541 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
      601 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
      661 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
      721 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
      781 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
      841 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
      901 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
      961 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1021 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1081 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1141 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1201 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1261 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1321 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1381 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     1441 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     1501 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     1561 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     1621 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     1681 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     1741 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat
     1801 cggttaactt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     1861 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     1921 aattttatcg atcacgagac tagcctcgag aagcttgata tcgaattcca cggggttggg
     1981 gttgcgcctt ttccaaggca gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg
     2041 ttccgggaaa cgcagcggcg ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag
     2101 cgtcacccgg atcttcgccg ctacccttgt gggccccccg gcgacgcttc ctgctccgcc
     2161 cctaagtcgg gaaggttcct tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc
     2221 acgtctcact agtaccctcg cagacggaca gcgccaggga gcaatggcag cgcgccgacc
     2281 gcgatgggct gtggccaata gcggctgctc agcagggcgc gccgagagca gcggccggga
     2341 aggggcggtg cgggaggcgg ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg
     2401 gtgttccgca ttctgcaagc ctccggagcg cacgtcggca gtcggctccc tcgttgaccg
     2461 aatcaccgac ctctctcccc agggggatcc gctatgagct acggccgccc ccctcccgat
     2521 gtggagggta tgacctccct caaggtggac aacctgacct accgcacctc gcccgacacg
     2581 ctgaggcgcg tcttcgagaa gtacgggcgc gtcggcgacg tgtacatccc gcgggatcgc
     2641 tacaccaagg agtcccgcgg cttcgccttc gttcgctttc acgacaagcg cgacgctgag
     2701 gacgctatgg atgccatgga cggggccgtg ctggacggcc gcgagctgcg ggtgcaaatg
     2761 gcgcgctacg gccgccaccc ggactcacac cacagccgcc ggggaccgcc accccgcagg
     2821 tacgggggcg gtggctacgg acgccggagc cgcagcccta ggcggcgtcg ccgcagccga
     2881 tcccggagtc ggagccgttc caggtctcgc agccgatctc gctacagccg ctcgaagtct
     2941 cggtcccgca ctcgttctcg atctcggtcg acctccaagt ccagatccgc acgaaggtcc
     3001 aagtccaagt cctcgtcggt ctccagatct cgttcgcggt ccaggtcccg gtctcggtcc
     3061 aggagtcctc ccccagtgtc caagagggaa tccaaatcca ggtcgcgatc gaagagtccc
     3121 cccaagtctc ctgaagagga aggagcggtg tcctctggcg gcgactacaa agacgatgac
     3181 gacaaggagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct
     3241 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
     3301 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
     3361 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
     3421 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
     3481 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
     3541 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
     3601 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
     3661 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
     3721 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
     3781 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
     3841 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
     3901 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta agcggccgcc
     3961 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     4021 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     4081 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     4141 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     4201 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     4261 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     4321 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     4381 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     4441 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     4501 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     4561 ttcgagctcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     4621 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct
     4681 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     4741 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     4801 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     4861 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     4921 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     4981 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5041 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     5101 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5161 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     5221 tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt
     5281 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     5341 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     5401 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     5461 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5521 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5581 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5641 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     5701 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5761 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     5821 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5881 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5941 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     6001 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     6061 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6121 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     6181 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     6241 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     6301 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     6361 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6421 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     6481 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     6541 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     6601 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     6661 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6721 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6781 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     6841 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     6901 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6961 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7021 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     7081 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     7141 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     7201 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     7261 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     7321 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     7381 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     7441 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     7501 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     7561 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     7621 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     7681 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     7741 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     7801 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     7861 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     7921 gggcagtgag cgcaacgcaa ttaaggtgag ttagctcact cattaggcac cccaggcttt
     7981 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     8041 aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca
     8101 aaagctggag ctgca
//
