LOCUS       sequence-232521-        4333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient transfection; Expresses RBM38-FRB; CAGGS promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4333)
  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 4333)
  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: pAC1809_pmax-RBM38-FRB
FEATURES             Location/Qualifiers
     source          1..4333
                     /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             1757..1777
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1781..1801
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1805..1825
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1907..2176
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQ"
     CDS             2279..2344
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(2363..2387)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2440..2481
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2507..2526)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    2536..2617
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2561..2580
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(2815..3403)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2895..2914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3484..4278)
                     /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(3484..4278)
                     /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(3595..3614)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4205..4224
                     /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 tgcttctgca
     1021 accggcgcca tgtgcacctt cagcaggttt tccaagaccg cttgctgcgc caggcgctat
     1081 gcacggcagc caaaaagaca ccacgtttac aaagatattc gtcggtggac tcccttatca
     1141 tactactgac gcgtccctta gaaagtactt cgagggtttc ggggatattg aagaagccgt
     1201 cgtcataacg gatcggcaaa ctggaaagag ccggggctac ggttttgtca cgatggctga
     1261 tagggcagcg gcagagaggg cttgtaaaga cccaaaccct attatcgatg ggcgcaaggc
     1321 aaacgtaaac ctggcatacc ttggggcaaa gccgcgctca ttgcagactg gcttcgctat
     1381 cggcgtacaa caactccacc caacattgat tcaaagaact tatggactca caccacatta
     1441 catttatccc ccagcaatag tacaaccatc agtcgtcatt cctgcagctc ccgttccatc
     1501 actctccagt ccttatatag agtatacacc agcctctcca gcttatgccc aatacccccc
     1561 tgccacttat gaccaatacc catacgccgc gagcccggcc accgcagcca gctttgtagg
     1621 ttatagttac ccggctgccg ttccccaagc tctcagcgct gccgctcctg ctggtactac
     1681 cttcgtacaa taccaggctc ctcagttgca gcccgacaga atgcaaaacg ttatcgatgg
     1741 cggaggcgga tctgatccaa aaaagaagag aaaggtagat ccaaaaaaga agagaaaggt
     1801 agatccaaaa aagaagagaa aggtaggatc taccggatct agaaacgatg gtggtggtgg
     1861 aagcgggggt gggggcagcg gtggaggggg aagcgggcgc gccatggaga tgtggcatga
     1921 aggcctggaa gaggcatctc gtttgtactt tggggaaagg aacgtgaaag gcatgtttga
     1981 ggtgctggag cccttgcatg ctatgatgga acggggcccc cagactctga aggaaacatc
     2041 ctttaatcag gcctatggtc gagatttaat ggaggcccaa gagtggtgca ggaagtacat
     2101 gaaatcaggg aatgtcaagg acctcaccca agcctgggac ctctattatc atgtgttccg
     2161 acgaatctca aagcagcaga tctcgtacgc gtcccggggc ggtggctcat ctggcggagg
     2221 tggaggcggc ggtagcggag gaggcgggtc cggcggcggc ggtagtgggc cggccatgga
     2281 ctacaaggat cacgacggtg actataagga tcatgacatc gactataagg acgatgacga
     2341 taagtgatta attaagaatt cgacccagct ttcttgtaca aagtggttga tatccagcac
     2401 agtggcggcc gctcgagtct agagggcccg cggttcgaag gtaagcctat ccctaaccct
     2461 ctcctcggtc tcgattctac gcgtaccggt tagcaattgt tttttcgatg agtttggaca
     2521 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2581 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2641 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     2701 atgtggtact taagaggggg agaccaaagg gcgagacgtt aaggcctcac gtgacatgtg
     2761 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2821 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2881 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2941 tgttccgacc ctgccgctta cgggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3001 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3061 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3121 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3181 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3241 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3301 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3361 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3421 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgcc
     3481 gtctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc
     3541 gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc
     3601 acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat
     3661 gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt
     3721 cacgacgaga tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca gttcggctgg
     3781 cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg
     3841 agtacgtgct ctctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc
     3901 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
     3961 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
     4021 ttcagtgaca acgtcgagta cagctgcgca aggaacgccc gtcgtggcca gccacgatag
     4081 ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
     4141 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
     4201 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
     4261 tccatcttgt tcaatcataa tattattgaa gcatttatca gggttcgtct cgtcccggtc
     4321 tcctcccatg cav
//
