LOCUS       sequence-232524-        4368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient transfection; Expresses RBFOX1N-FRB-C; CAGGS promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4368)
  AUTHORS   Du M, Jillette N, Zhu JJ, Li S, Cheng AW
  TITLE     CRISPR artificial splicing factors.
  JOURNAL   Nat Commun. 2020 Jun 12;11(1):2973. doi: 10.1038/s41467-020-16806-4.
  PUBMED    32532987
REFERENCE   2  (bases 1 to 4368)
  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: "Nat 
            Commun."; date: "2020-06-12"; volume: "11(1):2973. doi"; pages: " 
            10.1038/s41467-020-16806-4"
COMMENT     Sequence Label: pAC1808_pmax-RBFOX1N-FRB-C
FEATURES             Location/Qualifiers
     source          1..4368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        140..519
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        520..723
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     674..694
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          815..947
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     protein_bind    962..986
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1388..1657
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQ"
     protein_bind    complement(2399..2423)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2476..2517
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2543..2562)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    2572..2653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2597..2616
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(2851..3439)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2931..2950)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3520..4314)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(3520..4314)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3631..3650)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4241..4260
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tgtcaatatt ggccattagc catattattc attggttata tagcataaat caatattggc
       61 tattggccat tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt
      121 ccaatatgac cgccatgttg gcattgatta ttgactagtt attaatagta atcaattacg
      181 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      241 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      301 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      361 gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat tgacgtcaat
      421 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga ctttcctact
      481 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
      541 accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      601 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaataac
      661 cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      721 ggtcgtttag tgaaccgtca gatcactagt agctttattg cggtagttta tcacagttaa
      781 attgctaacg cagtcagtgc tcgactgatc acaggtaagt atcaaggtta caagacaggt
      841 ttaaggaggc caatagaaac tgggcttgtc gagacagaga agattcttgc gtttctgata
      901 ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggga aaaaacaatt
      961 gacaagtttg tacaaaaaag caggctccga attcaccggt gccgccacca tgaattgcga
     1021 gcgggaacag ctcaggggca atcaagaagc ggcagctgca cctgacacaa tggcacagcc
     1081 gtatgcgagt gctcaatttg cgccacccca gaatggcata cccgcagaat acacagcgcc
     1141 gcatcctcac cccgcccctg aatacactgg gcaaactacc gttcctgaac acacgcttaa
     1201 cctctatcct cctgctcaga ctcactcaga acagagtcca gcagatacaa gtgcgcagac
     1261 agtctctggc actgcgaccc agaccgatga cgcagctcct actgatggcc agccacaaac
     1321 tcaaccctct gaaaacactg aaaataaaag ccaacccaag ggcggtggag gctccgggcg
     1381 cgccatggag atgtggcatg aaggcctgga agaggcatct cgtttgtact ttggggaaag
     1441 gaacgtgaaa ggcatgtttg aggtgctgga gcccttgcat gctatgatgg aacggggccc
     1501 ccagactctg aaggaaacat cctttaatca ggcctatggt cgagatttaa tggaggccca
     1561 agagtggtgc aggaagtaca tgaaatcagg gaatgtcaag gacctcaccc aagcctggga
     1621 cctctattat catgtgttcc gacgaatctc aaagcagcag atctcgtacg cgtcccgggg
     1681 cggtggctca tctggcggag gtggaggcgg cggtagcgga ggaggcgggt ccggcggcgg
     1741 cggtagtggg ccggccaatg ccacggctag ggtcatgaca aacaagaaga cggttaatcc
     1801 gtataccaac gggtggaaac ttaatcccgt tgtcggcgct gtctattctc ccgagttcta
     1861 tgctggtaca gtcttgctgt gccaagccaa tcaagagggg tcttcaatgt acagtgctcc
     1921 gagcagcttg gtttacacaa gcgcgatgcc aggatttcct taccctgcgg caacggccgc
     1981 ggctgcgtat cggggtgccc accttcgcgg gcggggccga acagtttaca acacgttcag
     2041 ggccgctgct ccacctcccc cgattcccgc gtacggaggt gttgtctacc aagacgggtt
     2101 ttatggagcc gacatctacg gtggttacgc cgcatatcgg tatgcacaac caacccccgc
     2161 gaccgcggca gcgtacagcg actcctacgg acgcgtgtac gctgctgatc cttaccacca
     2221 cgccctggcc cccgccccaa cgtatggtgt cggtgctatg aatgcgttcg cgcccctgac
     2281 cgatgcaaaa accaggtcac atgccgacga cgttggattg gtattgagct ctttgcaagc
     2341 atccatttac cgggggggat acaaccgctt cgcgccttat tgattaatta agaattcgac
     2401 ccagctttct tgtacaaagt ggttgatatc cagcacagtg gcggccgctc gagtctagag
     2461 ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc tcggtctcga ttctacgcgt
     2521 accggttagc aattgttttt tcgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2581 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2641 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     2701 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtacttaag agggggagac
     2761 caaagggcga gacgttaagg cctcacgtga catgtgagca aaaggccagc aaaaggccag
     2821 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2881 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2941 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttacggg
     3001 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3061 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3121 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3181 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3241 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3301 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3361 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3421 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3481 gaacgaaaac tcacgttaag ggattttggt catgccgtct cagaagaact cgtcaagaag
     3541 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg
     3601 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg
     3661 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc
     3721 caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg
     3781 catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc
     3841 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctctct cgatgcgatg
     3901 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc
     3961 atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc
     4021 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagtacagc
     4081 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cttgcagttc
     4141 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag
     4201 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag
     4261 cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcataatatt
     4321 attgaagcat ttatcagggt tcgtctcgtc ccggtctcct cccatgca
//
