LOCUS       sequence-232546-        6848 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor; dCasRx-RBM38 ORF; No promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6848)
  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 6848)
  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: pAC1825_pCR8-dCasRx-RBM38
FEATURES             Location/Qualifiers
     source          1..6848
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(157..184)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(276..362)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     411..433
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     425..442
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     426..442
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    458..557
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      575..584
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             584..649
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             674..694
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             698..718
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             722..742
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             824..844
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             854..3751
                     /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             3761..3781
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3791..3817
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(4616..4715)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4732..4751)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4733..4751)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4756..4772)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4756..4772)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5203..5994
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      6087..6675
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6576..6595
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6829..6846
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
       61 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac
      121 gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc
      181 ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc
      241 cgttgcttca caacgatcaa atccgctccc ggcggatttg tcctactcag gagagcgttc
      301 accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt
      361 tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg
      421 acgttgtaaa acgacggcca gtcttaagct cgggccccaa ataatgattt tattttgact
      481 gatagtgacc tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt
      541 gtacaaaaaa gcaggctccg aattcaccgg tgccgccacc atggactaca aggatcacga
      601 cggtgactat aaggatcatg acatcgacta taaggacgat gacgataaga tcgatggcgg
      661 aggcggatct gatccaaaaa agaagagaaa ggtagatcca aaaaagaaga gaaaggtaga
      721 tccaaaaaag aagagaaagg taggatctac cggatctaga aacgatggtg gtggtggaag
      781 cgggggtggg ggcagcggtg gagggggaag cgggcgcgcc agccccaaga agaagagaaa
      841 ggtggaggcc agcatcgaaa aaaaaaagtc cttcgccaag ggcatgggcg tgaagtccac
      901 actcgtgtcc ggctccaaag tgtacatgac aaccttcgcc gaaggcagcg acgccaggct
      961 ggaaaagatc gtggagggcg acagcatcag gagcgtgaat gagggcgagg ccttcagcgc
     1021 tgaaatggcc gataaaaacg ccggctataa gatcggcaac gccaaattca gccatcctaa
     1081 gggctacgcc gtggtggcta acaaccctct gtatacagga cccgtccagc aggatatgct
     1141 cggcctgaag gaaactctgg aaaagaggta cttcggcgag agcgctgatg gcaatgacaa
     1201 tatttgtatc caggtgatcc ataacatcct ggacattgaa aaaatcctcg ccgaatacat
     1261 taccaacgcc gcctacgccg tcaacaatat ctccggcctg gataaggaca ttattggatt
     1321 cggcaagttc tccacagtgt atacctacga cgaattcaaa gaccccgagc accatagggc
     1381 cgctttcaac aataacgata agctcatcaa cgccatcaag gcccagtatg acgagttcga
     1441 caacttcctc gataacccca gactcggcta tttcggccag gcctttttca gcaaggaggg
     1501 cagaaattac atcatcaatt acggcaacga atgctatgac attctggccc tcctgagcgg
     1561 actggcgcac tgggtggtcg ctaacaacga agaagagtcc aggatctcca ggacctggct
     1621 ctacaacctc gataagaacc tcgacaacga atacatctcc accctcaact acctctacga
     1681 caggatcacc aatgagctga ccaactcctt ctccaagaac tccgccgcca acgtgaacta
     1741 tattgccgaa actctgggaa tcaaccctgc cgaattcgcc gaacaatatt tcagattcag
     1801 cattatgaaa gagcagaaaa acctcggatt caatatcacc aagctcaggg aagtgatgct
     1861 ggacaggaag gatatgtccg agatcaggaa aaatcataag gtgttcgact ccatcaggac
     1921 caaggtctac accatgatgg actttgtgat ttataggtat tacatcgaag aggatgccaa
     1981 ggtggctgcc gccaataagt ccctccccga taatgagaag tccctgagcg agaaggatat
     2041 ctttgtgatt aacctgaggg gctccttcaa cgacgaccag aaggatgccc tctactacga
     2101 tgaagctaat agaatttgga gaaagctcga aaatatcatg cacaacatca aggaatttag
     2161 gggaaacaag acaagagagt ataagaagaa ggacgcccct agactgccca gaatcctgcc
     2221 cgctggccgt gatgtttccg ccttcagcaa actcatgtat gccctgacca tgttcctgga
     2281 tggcaaggag atcaacgacc tcctgaccac cctgattaat aaattcgata acatccagag
     2341 cttcctgaag gtgatgcctc tcatcggagt caacgctaag ttcgtggagg aatacgcctt
     2401 tttcaaagac tccgccaaga tcgccgatga gctgaggctg atcaagtcct tcgctagaat
     2461 gggagaacct attgccgatg ccaggagggc catgtatatc gacgccatcc gtattttagg
     2521 aaccaacctg tcctatgatg agctcaaggc cctcgccgac accttttccc tggacgagaa
     2581 cggaaacaag ctcaagaaag gcaagcacgg catgagaaat ttcattatta ataacgtgat
     2641 cagcaataaa aggttccact acctgatcag atacggtgat cctgcccacc tccatgagat
     2701 cgccaaaaac gaggccgtgg tgaagttcgt gctcggcagg atcgctgaca tccagaaaaa
     2761 acagggccag aacggcaaga accagatcga caggtactac gaaacttgta tcggaaagga
     2821 taagggcaag agcgtgagcg aaaaggtgga cgctctcaca aagatcatca ccggaatgaa
     2881 ctacgaccaa ttcgacaaga aaaggagcgt cattgaggac accggcaggg aaaacgccga
     2941 gagggagaag tttaaaaaga tcatcagcct gtacctcacc gtgatctacc acatcctcaa
     3001 gaatattgtc aatatcaacg ccaggtacgt catcggattc cattgcgtcg agcgtgatgc
     3061 tcaactgtac aaggagaaag gctacgacat caatctcaag aaactggaag agaagggatt
     3121 cagctccgtc accaagctct gcgctggcat tgatgaaact gcccccgata agagaaagga
     3181 cgtggaaaag gagatggctg aaagagccaa ggagagcatt gacagcctcg agagcgccaa
     3241 ccccaagctg tatgccaatt acatcaaata cagcgacgag aagaaagccg aggagttcac
     3301 caggcagatt aacagggaga aggccaaaac cgccctgaac gcctacctga ggaacaccaa
     3361 gtggaatgtg atcatcaggg aggacctcct gagaattgac aacaagacat gtaccctgtt
     3421 cgcaaacaag gccgtcgccc tggaagtggc caggtatgtc cacgcctata tcaacgacat
     3481 tgccgaggtc aattcctact tccaactgta ccattacatc atgcagagaa ttatcatgaa
     3541 tgagaggtac gagaaaagca gcggaaaggt gtccgagtac ttcgacgctg tgaatgacga
     3601 gaagaagtac aacgataggc tcctgaaact gctgtgtgtg cctttcggct actgtatccc
     3661 caggtttaag aacctgagca tcgaggccct gttcgatagg aacgaggccg ccaagttcga
     3721 caaggagaaa aagaaggtgt ccggcaattc cggatccgga cctaagaaaa agaggaaggt
     3781 ggcggccgct tacccatacg atgttccaga ttacgctggc ggccgcgggg gaggcggttc
     3841 cggtggcggc ggaagcggag gtggaggatc agggccggcc atgcttctgc aaccggcgcc
     3901 atgtgcacct tcagcaggtt ttccaagacc gcttgctgcg ccaggcgcta tgcacggcag
     3961 ccaaaaagac accacgttta caaagatatt cgtcggtgga ctcccttatc atactactga
     4021 cgcgtccctt agaaagtact tcgagggttt cggggatatt gaagaagccg tcgtcataac
     4081 ggatcggcaa actggaaaga gccggggcta cggttttgtc acgatggctg atagggcagc
     4141 ggcagagagg gcttgtaaag acccaaaccc tattatcgat gggcgcaagg caaacgtaaa
     4201 cctggcatac cttggggcaa agccgcgctc attgcagact ggcttcgcta tcggcgtaca
     4261 acaactccac ccaacattga ttcaaagaac ttatggactc acaccacatt acatttatcc
     4321 cccagcaata gtacaaccat cagtcgtcat tcctgcagct cccgttccat cactctccag
     4381 tccttatata gagtatacac cagcctctcc agcttatgcc caataccccc ctgccaccta
     4441 tgaccaatac ccatacgccg cgagcccggc caccgcagcc agctttgtag gttatagtta
     4501 cccggctgcc gttccccaag ctctcagcgc tgccgctcct gctggtacta ccttcgtaca
     4561 ataccaggct cctcagttgc agcccgacag aatgcaatga ttaattaaga attcgaccca
     4621 gctttcttgt acaaagttgg cattataaaa aataattgct catcaatttg ttgcaacgaa
     4681 caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt
     4741 gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc
     4801 tgatgttaca ttgcacaaga taaaaatata tcatcatgcc tcctctagac cagccaggac
     4861 agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg
     4921 atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag
     4981 cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca
     5041 gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca
     5101 tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat
     5161 gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc
     5221 gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg
     5281 acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt
     5341 gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg
     5401 atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta
     5461 gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa
     5521 ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg
     5581 atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta
     5641 ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta
     5701 aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta
     5761 gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat
     5821 gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa
     5881 gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg
     5941 gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg
     6001 agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac
     6061 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6121 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6181 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6241 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6301 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6361 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6421 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6481 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6541 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6601 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6661 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6721 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6781 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     6841 agcgagga
//
