LOCUS       sequence-232478-        7020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient transfection; Expresses RBM38-dCasRx; CAGGS promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7020)
  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 7020)
  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: pAC1803_pmax-RBM38-dCasRx
FEATURES             Location/Qualifiers
     source          1..7020
                     /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..1927
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1937..4834
                     /codon_start=1
                     /product="RNA-targeting Type VI CRISPR protein from 
                     Ruminococcus flavefaciens strain XPD3002 (Konermann et al.,
                     2018)"
                     /label=RfxCas13d
                     /translation="IEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDSI
                     RSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLEK
                     RYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFSTVY
                     TYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYIIN
                     YGNECYDILALLSGLAHWVVANNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNE
                     LTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKD
                     MSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVI
                     NLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPAG
                     RDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAFF
                     KDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDEN
                     GNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQK
                     KQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGREN
                     AEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLEE
                     KGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKKA
                     EEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFANKAVALEVARYVHA
                     YINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVP
                     FGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS"
     CDS             4844..4864
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4874..4900
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4967..5032
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(5051..5075)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5128..5169
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(5195..5214)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    5224..5305
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     5249..5268
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5503..6091)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5583..5602)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6172..6966)
                     /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(6172..6966)
                     /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(6283..6302)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6893..6912
                     /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 gccagcccca agaagaagag
     1921 aaaggtggag gccagcatcg aaaaaaaaaa gtccttcgcc aagggcatgg gcgtgaagtc
     1981 cacactcgtg tccggctcca aagtgtacat gacaaccttc gccgaaggca gcgacgccag
     2041 gctggaaaag atcgtggagg gcgacagcat caggagcgtg aatgagggcg aggccttcag
     2101 cgctgaaatg gccgataaaa acgccggcta taagatcggc aacgccaaat tcagccatcc
     2161 taagggctac gccgtggtgg ctaacaaccc tctgtataca ggacccgtcc agcaggatat
     2221 gctcggcctg aaggaaactc tggaaaagag gtacttcggc gagagcgctg atggcaatga
     2281 caatatttgt atccaggtga tccataacat cctggacatt gaaaaaatcc tcgccgaata
     2341 cattaccaac gccgcctacg ccgtcaacaa tatctccggc ctggataagg acattattgg
     2401 attcggcaag ttctccacag tgtataccta cgacgaattc aaagaccccg agcaccatag
     2461 ggccgctttc aacaataacg ataagctcat caacgccatc aaggcccagt atgacgagtt
     2521 cgacaacttc ctcgataacc ccagactcgg ctatttcggc caggcctttt tcagcaagga
     2581 gggcagaaat tacatcatca attacggcaa cgaatgctat gacattctgg ccctcctgag
     2641 cggactggcg cactgggtgg tcgctaacaa cgaagaagag tccaggatct ccaggacctg
     2701 gctctacaac ctcgataaga acctcgacaa cgaatacatc tccaccctca actacctcta
     2761 cgacaggatc accaatgagc tgaccaactc cttctccaag aactccgccg ccaacgtgaa
     2821 ctatattgcc gaaactctgg gaatcaaccc tgccgaattc gccgaacaat atttcagatt
     2881 cagcattatg aaagagcaga aaaacctcgg attcaatatc accaagctca gggaagtgat
     2941 gctggacagg aaggatatgt ccgagatcag gaaaaatcat aaggtgttcg actccatcag
     3001 gaccaaggtc tacaccatga tggactttgt gatttatagg tattacatcg aagaggatgc
     3061 caaggtggct gccgccaata agtccctccc cgataatgag aagtccctga gcgagaagga
     3121 tatctttgtg attaacctga ggggctcctt caacgacgac cagaaggatg ccctctacta
     3181 cgatgaagct aatagaattt ggagaaagct cgaaaatatc atgcacaaca tcaaggaatt
     3241 taggggaaac aagacaagag agtataagaa gaaggacgcc cctagactgc ccagaatcct
     3301 gcccgctggc cgtgatgttt ccgccttcag caaactcatg tatgccctga ccatgttcct
     3361 ggatggcaag gagatcaacg acctcctgac caccctgatt aataaattcg ataacatcca
     3421 gagcttcctg aaggtgatgc ctctcatcgg agtcaacgct aagttcgtgg aggaatacgc
     3481 ctttttcaaa gactccgcca agatcgccga tgagctgagg ctgatcaagt ccttcgctag
     3541 aatgggagaa cctattgccg atgccaggag ggccatgtat atcgacgcca tccgtatttt
     3601 aggaaccaac ctgtcctatg atgagctcaa ggccctcgcc gacacctttt ccctggacga
     3661 gaacggaaac aagctcaaga aaggcaagca cggcatgaga aatttcatta ttaataacgt
     3721 gatcagcaat aaaaggttcc actacctgat cagatacggt gatcctgccc acctccatga
     3781 gatcgccaaa aacgaggccg tggtgaagtt cgtgctcggc aggatcgctg acatccagaa
     3841 aaaacagggc cagaacggca agaaccagat cgacaggtac tacgaaactt gtatcggaaa
     3901 ggataagggc aagagcgtga gcgaaaaggt ggacgctctc acaaagatca tcaccggaat
     3961 gaactacgac caattcgaca agaaaaggag cgtcattgag gacaccggca gggaaaacgc
     4021 cgagagggag aagtttaaaa agatcatcag cctgtacctc accgtgatct accacatcct
     4081 caagaatatt gtcaatatca acgccaggta cgtcatcgga ttccattgcg tcgagcgtga
     4141 tgctcaactg tacaaggaga aaggctacga catcaatctc aagaaactgg aagagaaggg
     4201 attcagctcc gtcaccaagc tctgcgctgg cattgatgaa actgcccccg ataagagaaa
     4261 ggacgtggaa aaggagatgg ctgaaagagc caaggagagc attgacagcc tcgagagcgc
     4321 caaccccaag ctgtatgcca attacatcaa atacagcgac gagaagaaag ccgaggagtt
     4381 caccaggcag attaacaggg agaaggccaa aaccgccctg aacgcctacc tgaggaacac
     4441 caagtggaat gtgatcatca gggaggacct cctgagaatt gacaacaaga catgtaccct
     4501 gttcgcaaac aaggccgtcg ccctggaagt ggccaggtat gtccacgcct atatcaacga
     4561 cattgccgag gtcaattcct acttccaact gtaccattac atcatgcaga gaattatcat
     4621 gaatgagagg tacgagaaaa gcagcggaaa ggtgtccgag tacttcgacg ctgtgaatga
     4681 cgagaagaag tacaacgata ggctcctgaa actgctgtgt gtgcctttcg gctactgtat
     4741 ccccaggttt aagaacctga gcatcgaggc cctgttcgat aggaacgagg ccgccaagtt
     4801 cgacaaggag aaaaagaagg tgtccggcaa ttccggatcc ggacctaaga aaaagaggaa
     4861 ggtggcggcc gcttacccat acgatgttcc agattacgct ggcggccgcg ggggaggcgg
     4921 ttccggtggc ggcggaagcg gaggtggagg atcagggccg gccatggact acaaggatca
     4981 cgacggtgac tataaggatc atgacatcga ctataaggac gatgacgata agtgattaat
     5041 taagaattcg acccagcttt cttgtacaaa gtggttgata tccagcacag tggcggccgc
     5101 tcgagtctag agggcccgcg gttcgaaggt aagcctatcc ctaaccctct cctcggtctc
     5161 gattctacgc gtaccggtta gcaattgttt tttcgatgag tttggacaaa ccacaactag
     5221 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     5281 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     5341 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtactta
     5401 agagggggag accaaagggc gagacgttaa ggcctcacgt gacatgtgag caaaaggcca
     5461 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5521 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5581 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5641 gccgcttacg ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5701 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5761 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5821 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5881 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5941 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6001 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6061 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6121 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgccgt ctcagaagaa
     6181 ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag
     6241 cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa
     6301 cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa
     6361 gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc
     6421 ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg
     6481 atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctct
     6541 ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag
     6601 ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag
     6661 gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac
     6721 gtcgagtaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc
     6781 gtcttgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc
     6841 ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc
     6901 atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc
     6961 aatcataata ttattgaagc atttatcagg gttcgtctcg tcccggtctc ctcccatgca
//
