LOCUS       sequence-232445-        7014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient transfection; Expresses dCasRx-RBM38; CAGGS promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7014)
  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 7014)
  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: pAC1804_pmax-dCasRx-RBM38
FEATURES             Location/Qualifiers
     source          1..7014
                     /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"
     regulatory      1004..1013
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1013..1078
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1103..1123
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1127..1147
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1151..1171
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1253..1273
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1283..4180
                     /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             4190..4210
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4220..4246
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(5045..5069)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5122..5163
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(5189..5208)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    5218..5299
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     5243..5262
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5497..6085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5577..5596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6166..6960)
                     /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(6166..6960)
                     /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(6277..6296)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6887..6906
                     /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 tggactacaa
     1021 ggatcacgac ggtgactata aggatcatga catcgactat aaggacgatg acgataagat
     1081 cgatggcgga ggcggatctg atccaaaaaa gaagagaaag gtagatccaa aaaagaagag
     1141 aaaggtagat ccaaaaaaga agagaaaggt aggatctacc ggatctagaa acgatggtgg
     1201 tggtggaagc gggggtgggg gcagcggtgg agggggaagc gggcgcgcca gccccaagaa
     1261 gaagagaaag gtggaggcca gcatcgaaaa aaaaaagtcc ttcgccaagg gcatgggcgt
     1321 gaagtccaca ctcgtgtccg gctccaaagt gtacatgaca accttcgccg aaggcagcga
     1381 cgccaggctg gaaaagatcg tggagggcga cagcatcagg agcgtgaatg agggcgaggc
     1441 cttcagcgct gaaatggccg ataaaaacgc cggctataag atcggcaacg ccaaattcag
     1501 ccatcctaag ggctacgccg tggtggctaa caaccctctg tatacaggac ccgtccagca
     1561 ggatatgctc ggcctgaagg aaactctgga aaagaggtac ttcggcgaga gcgctgatgg
     1621 caatgacaat atttgtatcc aggtgatcca taacatcctg gacattgaaa aaatcctcgc
     1681 cgaatacatt accaacgccg cctacgccgt caacaatatc tccggcctgg ataaggacat
     1741 tattggattc ggcaagttct ccacagtgta tacctacgac gaattcaaag accccgagca
     1801 ccatagggcc gctttcaaca ataacgataa gctcatcaac gccatcaagg cccagtatga
     1861 cgagttcgac aacttcctcg ataaccccag actcggctat ttcggccagg cctttttcag
     1921 caaggagggc agaaattaca tcatcaatta cggcaacgaa tgctatgaca ttctggccct
     1981 cctgagcgga ctggcgcact gggtggtcgc taacaacgaa gaagagtcca ggatctccag
     2041 gacctggctc tacaacctcg ataagaacct cgacaacgaa tacatctcca ccctcaacta
     2101 cctctacgac aggatcacca atgagctgac caactccttc tccaagaact ccgccgccaa
     2161 cgtgaactat attgccgaaa ctctgggaat caaccctgcc gaattcgccg aacaatattt
     2221 cagattcagc attatgaaag agcagaaaaa cctcggattc aatatcacca agctcaggga
     2281 agtgatgctg gacaggaagg atatgtccga gatcaggaaa aatcataagg tgttcgactc
     2341 catcaggacc aaggtctaca ccatgatgga ctttgtgatt tataggtatt acatcgaaga
     2401 ggatgccaag gtggctgccg ccaataagtc cctccccgat aatgagaagt ccctgagcga
     2461 gaaggatatc tttgtgatta acctgagggg ctccttcaac gacgaccaga aggatgccct
     2521 ctactacgat gaagctaata gaatttggag aaagctcgaa aatatcatgc acaacatcaa
     2581 ggaatttagg ggaaacaaga caagagagta taagaagaag gacgccccta gactgcccag
     2641 aatcctgccc gctggccgtg atgtttccgc cttcagcaaa ctcatgtatg ccctgaccat
     2701 gttcctggat ggcaaggaga tcaacgacct cctgaccacc ctgattaata aattcgataa
     2761 catccagagc ttcctgaagg tgatgcctct catcggagtc aacgctaagt tcgtggagga
     2821 atacgccttt ttcaaagact ccgccaagat cgccgatgag ctgaggctga tcaagtcctt
     2881 cgctagaatg ggagaaccta ttgccgatgc caggagggcc atgtatatcg acgccatccg
     2941 tattttagga accaacctgt cctatgatga gctcaaggcc ctcgccgaca ccttttccct
     3001 ggacgagaac ggaaacaagc tcaagaaagg caagcacggc atgagaaatt tcattattaa
     3061 taacgtgatc agcaataaaa ggttccacta cctgatcaga tacggtgatc ctgcccacct
     3121 ccatgagatc gccaaaaacg aggccgtggt gaagttcgtg ctcggcagga tcgctgacat
     3181 ccagaaaaaa cagggccaga acggcaagaa ccagatcgac aggtactacg aaacttgtat
     3241 cggaaaggat aagggcaaga gcgtgagcga aaaggtggac gctctcacaa agatcatcac
     3301 cggaatgaac tacgaccaat tcgacaagaa aaggagcgtc attgaggaca ccggcaggga
     3361 aaacgccgag agggagaagt ttaaaaagat catcagcctg tacctcaccg tgatctacca
     3421 catcctcaag aatattgtca atatcaacgc caggtacgtc atcggattcc attgcgtcga
     3481 gcgtgatgct caactgtaca aggagaaagg ctacgacatc aatctcaaga aactggaaga
     3541 gaagggattc agctccgtca ccaagctctg cgctggcatt gatgaaactg cccccgataa
     3601 gagaaaggac gtggaaaagg agatggctga aagagccaag gagagcattg acagcctcga
     3661 gagcgccaac cccaagctgt atgccaatta catcaaatac agcgacgaga agaaagccga
     3721 ggagttcacc aggcagatta acagggagaa ggccaaaacc gccctgaacg cctacctgag
     3781 gaacaccaag tggaatgtga tcatcaggga ggacctcctg agaattgaca acaagacatg
     3841 taccctgttc gcaaacaagg ccgtcgccct ggaagtggcc aggtatgtcc acgcctatat
     3901 caacgacatt gccgaggtca attcctactt ccaactgtac cattacatca tgcagagaat
     3961 tatcatgaat gagaggtacg agaaaagcag cggaaaggtg tccgagtact tcgacgctgt
     4021 gaatgacgag aagaagtaca acgataggct cctgaaactg ctgtgtgtgc ctttcggcta
     4081 ctgtatcccc aggtttaaga acctgagcat cgaggccctg ttcgatagga acgaggccgc
     4141 caagttcgac aaggagaaaa agaaggtgtc cggcaattcc ggatccggac ctaagaaaaa
     4201 gaggaaggtg gcggccgctt acccatacga tgttccagat tacgctggcg gccgcggggg
     4261 aggcggttcc ggtggcggcg gaagcggagg tggaggatca gggccggcca tgcttctgca
     4321 accggcgcca tgtgcacctt cagcaggttt tccaagaccg cttgctgcgc caggcgctat
     4381 gcacggcagc caaaaagaca ccacgtttac aaagatattc gtcggtggac tcccttatca
     4441 tactactgac gcgtccctta gaaagtactt cgagggtttc ggggatattg aagaagccgt
     4501 cgtcataacg gatcggcaaa ctggaaagag ccggggctac ggttttgtca cgatggctga
     4561 tagggcagcg gcagagaggg cttgtaaaga cccaaaccct attatcgatg ggcgcaaggc
     4621 aaacgtaaac ctggcatacc ttggggcaaa gccgcgctca ttgcagactg gcttcgctat
     4681 cggcgtacaa caactccacc caacattgat tcaaagaact tatggactca caccacatta
     4741 catttatccc ccagcaatag tacaaccatc agtcgtcatt cctgcagctc ccgttccatc
     4801 actctccagt ccttatatag agtatacacc agcctctcca gcttatgccc aatacccccc
     4861 tgccacctat gaccaatacc catacgccgc gagcccggcc accgcagcca gctttgtagg
     4921 ttatagttac ccggctgccg ttccccaagc tctcagcgct gccgctcctg ctggtactac
     4981 cttcgtacaa taccaggctc ctcagttgca gcccgacaga atgcaatgat taattaagaa
     5041 ttcgacccag ctttcttgta caaagtggtt gatatccagc acagtggcgg ccgctcgagt
     5101 ctagagggcc cgcggttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct
     5161 acgcgtaccg gttagcaatt gttttttcga tgagtttgga caaaccacaa ctagaatgca
     5221 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     5281 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     5341 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta cttaagaggg
     5401 ggagaccaaa gggcgagacg ttaaggcctc acgtgacatg tgagcaaaag gccagcaaaa
     5461 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5521 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5581 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5641 tacgggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5701 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5761 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5821 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5881 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5941 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6001 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6061 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6121 tcagtggaac gaaaactcac gttaagggat tttggtcatg ccgtctcaga agaactcgtc
     6181 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
     6241 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
     6301 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
     6361 attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc
     6421 gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc
     6481 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctctctcgat
     6541 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg
     6601 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc
     6661 ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag
     6721 tacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg
     6781 cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc
     6841 tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc
     6901 gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat
     6961 aatattattg aagcatttat cagggttcgt ctcgtcccgg tctcctccca tgca
//
