LOCUS       sequence-232472-        7056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient transfection; Expresses RBFOX1N-dCasRx-C; CAGGS promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7056)
  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 7056)
  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: pAC1802_pmax-RBFOX1N-dCasRx-C
FEATURES             Location/Qualifiers
     source          1..7056
                     /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..1408
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1418..4315
                     /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             4325..4345
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4355..4381
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(5087..5111)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5164..5205
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(5231..5250)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    5260..5341
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     5285..5304
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5539..6127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5619..5638)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6208..7002)
                     /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(6208..7002)
                     /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(6319..6338)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6929..6948
                     /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 cgccagcccc aagaagaaga gaaaggtgga ggccagcatc gaaaaaaaaa agtccttcgc
     1441 caagggcatg ggcgtgaagt ccacactcgt gtccggctcc aaagtgtaca tgacaacctt
     1501 cgccgaaggc agcgacgcca ggctggaaaa gatcgtggag ggcgacagca tcaggagcgt
     1561 gaatgagggc gaggccttca gcgctgaaat ggccgataaa aacgccggct ataagatcgg
     1621 caacgccaaa ttcagccatc ctaagggcta cgccgtggtg gctaacaacc ctctgtatac
     1681 aggacccgtc cagcaggata tgctcggcct gaaggaaact ctggaaaaga ggtacttcgg
     1741 cgagagcgct gatggcaatg acaatatttg tatccaggtg atccataaca tcctggacat
     1801 tgaaaaaatc ctcgccgaat acattaccaa cgccgcctac gccgtcaaca atatctccgg
     1861 cctggataag gacattattg gattcggcaa gttctccaca gtgtatacct acgacgaatt
     1921 caaagacccc gagcaccata gggccgcttt caacaataac gataagctca tcaacgccat
     1981 caaggcccag tatgacgagt tcgacaactt cctcgataac cccagactcg gctatttcgg
     2041 ccaggccttt ttcagcaagg agggcagaaa ttacatcatc aattacggca acgaatgcta
     2101 tgacattctg gccctcctga gcggactggc gcactgggtg gtcgctaaca acgaagaaga
     2161 gtccaggatc tccaggacct ggctctacaa cctcgataag aacctcgaca acgaatacat
     2221 ctccaccctc aactacctct acgacaggat caccaatgag ctgaccaact ccttctccaa
     2281 gaactccgcc gccaacgtga actatattgc cgaaactctg ggaatcaacc ctgccgaatt
     2341 cgccgaacaa tatttcagat tcagcattat gaaagagcag aaaaacctcg gattcaatat
     2401 caccaagctc agggaagtga tgctggacag gaaggatatg tccgagatca ggaaaaatca
     2461 taaggtgttc gactccatca ggaccaaggt ctacaccatg atggactttg tgatttatag
     2521 gtattacatc gaagaggatg ccaaggtggc tgccgccaat aagtccctcc ccgataatga
     2581 gaagtccctg agcgagaagg atatctttgt gattaacctg aggggctcct tcaacgacga
     2641 ccagaaggat gccctctact acgatgaagc taatagaatt tggagaaagc tcgaaaatat
     2701 catgcacaac atcaaggaat ttaggggaaa caagacaaga gagtataaga agaaggacgc
     2761 ccctagactg cccagaatcc tgcccgctgg ccgtgatgtt tccgccttca gcaaactcat
     2821 gtatgccctg accatgttcc tggatggcaa ggagatcaac gacctcctga ccaccctgat
     2881 taataaattc gataacatcc agagcttcct gaaggtgatg cctctcatcg gagtcaacgc
     2941 taagttcgtg gaggaatacg cctttttcaa agactccgcc aagatcgccg atgagctgag
     3001 gctgatcaag tccttcgcta gaatgggaga acctattgcc gatgccagga gggccatgta
     3061 tatcgacgcc atccgtattt taggaaccaa cctgtcctat gatgagctca aggccctcgc
     3121 cgacaccttt tccctggacg agaacggaaa caagctcaag aaaggcaagc acggcatgag
     3181 aaatttcatt attaataacg tgatcagcaa taaaaggttc cactacctga tcagatacgg
     3241 tgatcctgcc cacctccatg agatcgccaa aaacgaggcc gtggtgaagt tcgtgctcgg
     3301 caggatcgct gacatccaga aaaaacaggg ccagaacggc aagaaccaga tcgacaggta
     3361 ctacgaaact tgtatcggaa aggataaggg caagagcgtg agcgaaaagg tggacgctct
     3421 cacaaagatc atcaccggaa tgaactacga ccaattcgac aagaaaagga gcgtcattga
     3481 ggacaccggc agggaaaacg ccgagaggga gaagtttaaa aagatcatca gcctgtacct
     3541 caccgtgatc taccacatcc tcaagaatat tgtcaatatc aacgccaggt acgtcatcgg
     3601 attccattgc gtcgagcgtg atgctcaact gtacaaggag aaaggctacg acatcaatct
     3661 caagaaactg gaagagaagg gattcagctc cgtcaccaag ctctgcgctg gcattgatga
     3721 aactgccccc gataagagaa aggacgtgga aaaggagatg gctgaaagag ccaaggagag
     3781 cattgacagc ctcgagagcg ccaaccccaa gctgtatgcc aattacatca aatacagcga
     3841 cgagaagaaa gccgaggagt tcaccaggca gattaacagg gagaaggcca aaaccgccct
     3901 gaacgcctac ctgaggaaca ccaagtggaa tgtgatcatc agggaggacc tcctgagaat
     3961 tgacaacaag acatgtaccc tgttcgcaaa caaggccgtc gccctggaag tggccaggta
     4021 tgtccacgcc tatatcaacg acattgccga ggtcaattcc tacttccaac tgtaccatta
     4081 catcatgcag agaattatca tgaatgagag gtacgagaaa agcagcggaa aggtgtccga
     4141 gtacttcgac gctgtgaatg acgagaagaa gtacaacgat aggctcctga aactgctgtg
     4201 tgtgcctttc ggctactgta tccccaggtt taagaacctg agcatcgagg ccctgttcga
     4261 taggaacgag gccgccaagt tcgacaagga gaaaaagaag gtgtccggca attccggatc
     4321 cggacctaag aaaaagagga aggtggcggc cgcttaccca tacgatgttc cagattacgc
     4381 tggcggccgc gggggaggcg gttccggtgg cggcggaagc ggaggtggag gatcagggcc
     4441 ggccaatgcc acggctaggg tcatgacaaa caagaagacg gttaatccgt ataccaacgg
     4501 gtggaaactt aatcccgttg tcggcgctgt ctattctccc gagttctatg ctggtacagt
     4561 cttgctgtgc caagccaatc aagaggggtc ttcaatgtac agtgctccga gcagcttggt
     4621 ttacacaagc gcgatgccag gatttcctta ccctgcggca acggccgcgg ctgcgtatcg
     4681 gggtgcccac cttcgcgggc ggggccgaac agtttacaac acgttcaggg ccgctgctcc
     4741 acctcccccg attcccgcgt acggaggtgt tgtctaccaa gacgggtttt atggagccga
     4801 catctacggt ggttacgccg catatcggta tgcacaacca acccccgcga ccgcggcagc
     4861 gtacagcgac tcctacggac gcgtgtacgc tgctgatcct taccaccacg ccctggcccc
     4921 cgccccaacg tatggtgtcg gtgctatgaa tgcgttcgcg cccctgaccg atgcaaaaac
     4981 caggtcacat gccgacgacg ttggattggt attgagctct ttgcaagcat ccatttaccg
     5041 ggggggatac aaccgcttcg cgccttattg attaattaag aattcgaccc agctttcttg
     5101 tacaaagtgg ttgatatcca gcacagtggc ggccgctcga gtctagaggg cccgcggttc
     5161 gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggttagcaa
     5221 ttgttttttc gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     5281 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     5341 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     5401 ttaaagcaag taaaacctct acaaatgtgg tacttaagag ggggagacca aagggcgaga
     5461 cgttaaggcc tcacgtgaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5521 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5581 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5641 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttacgggat acctgtccgc
     5701 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5761 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5821 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5881 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5941 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6001 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6061 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6121 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6181 acgttaaggg attttggtca tgccgtctca gaagaactcg tcaagaaggc gatagaaggc
     6241 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc
     6301 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc
     6361 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt
     6421 cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt
     6481 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg
     6541 atcgacaaga ccggcttcca tccgagtacg tgctctctcg atgcgatgtt tcgcttggtg
     6601 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat
     6661 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc
     6721 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agtacagctg cgcaaggaac
     6781 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc
     6841 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc
     6901 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca
     6961 agcggccgga gaacctgcgt gcaatccatc ttgttcaatc ataatattat tgaagcattt
     7021 atcagggttc gtctcgtccc ggtctcctcc catgca
//
