LOCUS       sequence-433911-        5410 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3-exon minigene, with SRSF1 motif-rich middle exon, hnRNPA1 
            motif-rich region in second intron, both randomly mutated to avoid 
            sequence repeats, and AG>GG mutation in final 3' splice site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5410)
  TITLE     RNA molecules display distinctive organization at nuclear speckles
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5410)
  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            "Unpublished"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: MUT_S1-H1,NR
FEATURES             Location/Qualifiers
     source          1..5410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(19..37,38..60,61..79,80..102,103..121,122..144,
                     145..163,164..186,187..205,206..228,229..247,248..270,
                     271..289)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        322..360
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     357..381
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     polyA_signal    2740..2874
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2764..2783)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2818..2837
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3307..3326)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(3420..3437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3591..4179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3671..3690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4350..5141,5142..5210))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4973..4992
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5211..5315)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5383..5401
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttcactcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac
       61 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
      121 aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt
      181 taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg
      241 atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
      301 cgagctcggt acccgggtcg aggtaggcgt gtacggtggg aggcctatat aagcagagct
      361 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
      421 agacaccggg accgatccag cctccgcgta cggttcgacc gctgaactgc atcgtcgccg
      481 tgtcccagaa taagggcatc ggcaagaacg gagaccttcc ctggccaaag ctcaggtact
      541 ggctggattg ggttagggaa accgaggcgg ttcgctgaat cgggtcgagc acttggcgga
      601 gacgcgcggg ccaactactt agggacagtc atgaggggta ggcccgccgg ctgcagccct
      661 tgcccatgcc cgcggtgatc cccatgctgt gccagccttt gcccagaggc gctctagctg
      721 ggagcaaagt ccggtcactg ggcagcacca ccccccggac ttgcatgggt agccgctgag
      781 atggagcctg agcacacgcg gccgctgtta acgcagtgtt tctctttttt tcaggccata
      841 caaggggaat agcgagcagg aggaaagcgc cgcaagacga gcaccaaaca ggagtagcgc
      901 cagtcaagga gaaggcaaat aaagaggaaa ctcaagcgaa tggaatggca cactaagcga
      961 acgccgagac acagcaacgt ttaactaccg cgacgtcaat caagacgatg ggcagcatcg
     1021 aggcatttca aacaacgtgt acgccagaca agcggatgtc gaagaccgag gaccgcccag
     1081 caacacaggt aagtctatgc gaacagggcc ccacgcagtt agtgtaacaa acaatagggt
     1141 cgggaaatta ttagggactg atgcaattag gctaacaaac aagtagggat cctactaatt
     1201 agggtaccaa tcaatttggg caccatgcaa ttagataccc aacctagtcg ctaatcaaac
     1261 aattatggaa gcacataata agggatccaa acgagtagga aaacaataaa ttaggcaatc
     1321 tcaggataag tgaaccatat gataagggca actaaaaagg cagagaccaa tccgcaagtg
     1381 gagtaaacaa ttaaggatcg actaatttag ggccaccaag agttagggta ccccagcatt
     1441 attaatcccc ggaattgggg aacgagagag ctaggaaaca acataagcta gctggccagt
     1501 gagatccaag aatcttcctg tctctgctga tccactgata ggattacaag tacatgccac
     1561 caagcccagc ttcctcttac catctgctgg ggaccaaact taggccctca ttcctacaca
     1621 gtgaatactt gactttgtta tcacccaacc ctaataaata actcactatc caaacaagtt
     1681 gaaaccctta gaattctgtg ttgctccagc atgatgttgt ggtaaacgtt aatacaataa
     1741 gatgcacagg tcataagtgc acattagcta agtgttgaca aagacttaga cctacataac
     1801 ttaaccctat tagccctcca gaaagttcct cattctccat tccaggcaac tttcatcaca
     1861 ccacatcatg tacaactact attgaagttg ttttccacta tagatacaat gagatgtcac
     1921 atacggcttt gtgttttgat ttgcaagtac caatcgagta tgaaatatgg agtggatatt
     1981 ggacattggc caccatctaa atactttgtg ttaaaagaat tggttttcat aatttgtttt
     2041 gtactgactg ctggctagtc agattacctg actagtatgg acaggatttt gcaataatca
     2101 taattctttt ttcggggaac caccacaagg agctcatttt cttgccaaaa gtctggacga
     2161 agccttaaaa cttattgaac aaccagagtt agcagataaa gtggagctgt catggtttgg
     2221 atagttggag gcagttccgt ttacaaggaa gccatgaatc agccaggcca tctcagactc
     2281 tttgtgacaa ggatcatgca ggaatttgaa agtgacacgt tcttcccaga aattgatttg
     2341 gagaaatata aacttctccc agagtaccca ggggtccttt ctgaagtcca ggaggaaaaa
     2401 ggcatcaagt ataaatttga agtctatgag aagaaaggct aacagaaaga tacttgctga
     2461 ttgacttcaa gttctactgc tttcctccta aaattatgca tttttacaag accatgggac
     2521 ttgtgttggc tttagatcta tgagttattc tttctttaga gagggatagt taggaagatg
     2581 tatttgtttt gtggtaccag agatggaacc tgggatcctg tgcatcctgg gcaactgttg
     2641 tactctaagc cactccccaa agtcatgccc cagcccctgt ataattctaa acaattagaa
     2701 ttattttcat tttcattagt ctaaccaggt tatctagagg atccagacat gataagatac
     2761 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2821 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2881 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     2941 aagtaaaacc tctacaaatg tggtatggct gattatgatc ctgcaagcct cgtcgtcctg
     3001 gccggaccac gctatctgtg caaggtcccc ggccccggac gcgcgctcca tgagcagagc
     3061 gcccgccgcc gaggcgaaga ctcgggcggc gccctgcccg tcccaccagg tcaacaggcg
     3121 gtaaccggcc tcttcatcgg gaatgcgcgc cttcagcatt gccggcatgt ccccctggcg
     3181 gacgggaagt atccagctcg accaagcttg gcgagatttt caggagctaa ggaagctaaa
     3241 atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa
     3301 cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt tcagctgcat
     3361 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3421 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3481 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3541 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3601 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3661 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3721 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3781 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3841 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3901 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3961 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4021 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4081 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4141 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4201 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4261 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4321 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4381 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4441 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4501 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4561 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4621 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4681 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4741 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4801 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4861 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4921 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc
     4981 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5041 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5101 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5161 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5221 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5281 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5341 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5401 ccctttcgtc
//
