LOCUS       sequence-472774-        4616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for constitutive mammalian expression of Flag-tagged 
            full-length SRSF3 mutated to be resistant to SRSF3 siRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4616)
  TITLE     A Scalable Exon Deletion Platform Coupled To Single-Cell 
            Transcriptomics Reveals Exons Shaping Gene Expression Dynamics
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4616)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCS2+ Flag-SRSF3_WobbleMut
FEATURES             Location/Qualifiers
     source          1..4616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     157..176
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(395..499)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(525..980)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(662..683)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     874..893
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1051..2035
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        2070..2088
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2070..2087
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      2120..2129
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2129..2152
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2703..2837
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2727..2746)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2781..2800
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2950..2970)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2950..2968)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2989..3005)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2989..3005)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3002..3024)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3013..3029
                     /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(3037..3043,3044..3061,3062..3067))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3082..3103
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3220..3237)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3391..3979)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3471..3490)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4150..4616,1..325,326..394))
                     /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"
ORIGIN
        1 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
       61 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
      121 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
      181 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
      241 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
      301 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
      361 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
      421 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
      481 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
      541 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
      601 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
      661 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
      721 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
      781 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
      841 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
      901 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
      961 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
     1021 atcggtgcgg gcctcttcgc tattacgcca gtcgaccgcc aattcaatat ggcgtatatg
     1081 gactcatgcc aattcaatat ggtggatctg gacctgtgcc aattcaatat ggcgtatatg
     1141 gactcgtgcc aattcaatat ggtggatctg gaccccagcc aattcaatat ggcggacttg
     1201 gcaccatgcc aattcaatat ggcggacctg gcactgtgcc aactggggag gggtctactt
     1261 ggcacggtgc caagtttgag gaggggtctt ggccctgtgc caagtccgcc atattgaatt
     1321 ggcatggtgc caataatggc ggccatattg gctatatgcc aggatcaata tataggcaat
     1381 atccaatatg gccctatgcc aatatggcta ttggccaggt tcaatactat gtattggccc
     1441 tatgccatat agtattccat atatgggttt tcctattgac gtagatagcc cctcccaatg
     1501 ggcggtccca tataccatat atggggcttc ctaataccgc ccatagccac tcccccattg
     1561 acgtcaatgg tctctatata tggtctttcc tattgacgtc atatgggcgg tcctattgac
     1621 gtatatggcg cctcccccat tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc
     1681 cattgacgtc aataggacca cccaccattg acgtcaatgg gatggctcat tgcccattca
     1741 tatccgttct cacgccccct attgacgtca atgacggtaa atggcccact tggcagtaca
     1801 tcaatatcta ttaatagtaa cttggcaagt acattactat tggaagtacg ccagggtaca
     1861 ttggcagtac tcccattgac gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca
     1921 tccccattga cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat
     1981 gggcggtagg cgtgcctaat gggaggtcta tataagcaat gctcgtttag ggaaccgcca
     2041 ttctgcctgg ggacgtcgga gcaagcttga tttaggtgac actatagaat acaagctact
     2101 tgttcttttt gcaggatcag ccaccatgga ttacaaggac gacgacgata agggatccca
     2161 tcgattcgaa ttccatcgtg attcctgtcc attggactgt aaggtttatg taggcaatct
     2221 tggaaacaat ggaaataaaa ccgagctgga gcgggctttt ggctactatg gaccactccg
     2281 aagtgtgtgg gttgctagaa acccacccgg ctttgctttt gttgaatttg aagatccccg
     2341 agatgcagct gatgcagtcc gagagctaga tggaagaaca ctatgtggct gccgtgtaag
     2401 agtggaactg tcgaatggtg aaaaaagaag tagaaatcgt ggcccacctc cctcttgggg
     2461 tcgtcgccct cgagatgatt atcgtaggag gagtcctcca cctcgtcgca gatctccaag
     2521 aaggagaagc ttctctcgca gccggagcag gtccctttct agagatagga gaagagagag
     2581 atcgctgtct cgggagagaa atcacaagcc gtcccgatcc ttctctaggt ctcgtagtcg
     2641 atctaggtca aatgaaagga aataactcga gcctctagaa ctatagtgag tcgtattacg
     2701 tagatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2761 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2821 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2881 aggtgtggga ggttttttaa ttcgcggccg cggcgccaat gcattgggcc cggtacccag
     2941 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     3001 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg ggagcataaa
     3061 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3121 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3181 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3241 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3301 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3361 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3421 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3481 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3541 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3601 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3661 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3721 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3781 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3841 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3901 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3961 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4021 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4081 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4141 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4201 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4261 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4321 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4381 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4441 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4501 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4561 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccg
//
