LOCUS       sequence-380384-        6881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express DjCas13d.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6881)
  AUTHORS   Back F, Sandoval A, Vu LM, Hong VM, Bhaskara A, Rodriguez SR, 
            O'Brien JT, Kolber BJ, Kroener S, Ploski JE
  TITLE     Adeno-associated viral vector resource for the RNA-targeting Cas13d:
            A comparison of high-fidelity variants, DjCas13d and hfCas13d.
  JOURNAL   Mol Ther Methods Clin Dev. 2025 Aug 20;33(4):101565. doi: 
            10.1016/j.omtm.2025.101565. eCollection 2025 Dec 11.
  PUBMED    41020266
REFERENCE   2  (bases 1 to 6881)
  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: "Mol Ther 
            Methods Clin Dev."; date: "2025-08-20"; volume: "33(4):101565. doi";
            pages: " 10.1016/j.omtm.2025.101565. eCollection 2025 Dec 11"
COMMENT     Sequence Label: pAAV-CMV-intron-DjCas13d-pA
FEATURES             Location/Qualifiers
     source          1..6881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     487..506
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   648..777
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        853..1156
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1157..1360
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1311..1331
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1357..1381
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1509..1887
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     CDS             1985..2005
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4655..4675
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4685..4711
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4726..4774
                     /note="synthetic polyadenylation signal"
     repeat_region   4791..4931
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4791..4920
                     /label=AAV2 ITR
     rep_origin      5006..5461
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5093..5112)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5303..5324
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5478..5497)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5597..5619
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5657..5675)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5743..5847
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5848..5916,5917..6708)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6066..6085)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6879..6881,1..586)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
       61 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
      121 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
      181 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
      241 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
      301 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
      361 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
      421 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
      481 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
      541 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
      601 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct
      661 gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct
      721 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg
      781 gccgcacgcg tctagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      841 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      901 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      961 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     1021 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     1081 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     1141 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     1201 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     1261 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     1321 cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat
     1381 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
     1441 cctccgcgga ttcgaatccc ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca
     1501 agagtgacgt aagtaccgcc tatagagtct ataggcccac aaaaaatgct ttcttctttt
     1561 aatatacttt tttgtttatc ttatttctaa tactttccct aatctctttc tttcagggca
     1621 ataatgatac aatgtatcat gcctctttgc accattctaa agaataacag tgataatttc
     1681 tgggttaagg caatagcaat atttctgcat ataaatattt ctgcatataa attgtaactg
     1741 atgtaagagg tttcatattg ctaatagcag ctacaatcca gctaccattc tgcttttatt
     1801 ttatggttgg gataaggctg gattattctg agtccaagct aggccctttt gctaatcatg
     1861 ttcatacctc ttatcttcct cccacagctc ctgggcaacg tgctggtctg tgtgctggcc
     1921 catcactttg gcaaagaatt gggattcgaa catcgatcgg accggtgcta gcgccaccat
     1981 gagccccaag aagaagagaa aggtggaggc cagcatcaag aagcaaaaat ccaagaaaac
     2041 ggtttctaaa acctccggat tgaaagaggc actttctgtg caggggactg tcataatgac
     2101 gtcttttggg aaggggaaca tggcgaatct ctcttacaag atcccgagct ctcaaaagcc
     2161 ccaaaatctc aatagtagcg ccggcttgaa gaacgtcgaa gtatctggta aaaagattaa
     2221 gttccaaggt cgacacccga agatcgccac taccgacaac cccctgttta agccgcagcc
     2281 aggtatggac cttctgtgcc tgaaagacaa actcgaaatg cactattttg ggaagacttt
     2341 cgacgacaat atccacatac aattgatata tcaaatactg gatattgaaa agatccttgc
     2401 ggttcatgtt aacaacatcg tattcacact ggacaatgtc ctgcatccgc aaaaggaaga
     2461 gctgactgaa gatttcattg gggcaggggg ctggcggatt aatctcgatt accaaactct
     2521 tagagggcag actaacaaat atgataggtt taaaaactac attaaaagaa aagaacttct
     2581 ttatttcggg gaggcatttt accatgaaaa cgagagacgg tatgaggaag acatcttcgc
     2641 aatacttacc ctgctgtctg cccttagaca gttttgcttt cattccgatc tgagtagtga
     2701 tgagagcgac cacgtgaatt cattctggct gtatcaactc gaggaccaac tgtctgacga
     2761 gttcaaggag actctcagta ttttgtggga agaggtcacc gaaaggatag atagtgaatt
     2821 ccttaaaact aatacagtaa acctccatat tttgtgtcat gttttcccga aagagagtaa
     2881 ggagacgatt gtacgcgctt attatgaatt cctgatcaag aagtccttta aaaatatggg
     2941 atttagcatc aagaaattgc gagagattat gctcgagcaa tcagatctta agagtttcaa
     3001 agaggataag tacaactccg tcagggctaa actgtataag ctcttcgact ttattataac
     3061 ctattactac gaccaccatg cttttgagaa ggaggcgctc gtgagctcct tgcgaagttc
     3121 actcactgag gaaaacaagg aagagattta catcaaaacg gcccggacac tcgcgagcgc
     3181 tcttggggcc gatttcaaga aagccgctgc cgatgtaaat gccaaaaaca taagggacta
     3241 tcagaagaag gcgaacgact ataggataag ttttgaagat attaaaatag ggaacacagg
     3301 cattgggtat ttctcagagc tcatatacat gctcaccctt ctcctcgatg gaaaggagat
     3361 aaacgacctc ctcacaaccc tgatcaacaa atttgataat ataatctcct tcatagacat
     3421 cctgaagaaa ctcaatttgg aatttaaatt caagcctgaa tacgcggact tctttaacat
     3481 gaccaactgc aggtatactc tcgaagagct gcgggttatc aacagtatcg ctagaatgca
     3541 aaagccaagc gccgacgccc gaaagatcat gtatcgcgat gctctgcgca tattgggaat
     3601 ggacaatcgg ccggatgaag agattgatcg ggaactcgaa aggacaatgc ccgtgggtgc
     3661 ggatggcaaa tttatcaaag gtaaacaggg ctttcgcaat ttcattgctt ccaacgttat
     3721 tgaatcttca cgcttccatt acctcgtacg gtataataat ccacacaaga cacggactct
     3781 tgtgaaaaac cctaatgtag taaaatttgt gctcgaaggg attccggaga cacaaataaa
     3841 acgatacttt gatgtttgta aaggtcaaga gatcccaccg acgtctgaca agtccgcaca
     3901 gatagacgtt cttgcccgaa taattagttc agtagactat aaaatatttg aagacgtacc
     3961 ccaaagtgca aagatcaata aagatgatcc tagccgaaac ttcagtgacg cgctgaaaaa
     4021 acagcggtac caggcaattg taagtcttta tttgactgtt atgtacctta taaccaaaaa
     4081 cctggtgtat gtcaatagta ggtatgtgat tgcatttcat tgcctggaac gagacgcatt
     4141 ccttcacggt gtgacgctcc ctaagatgaa taaaaaaatt gtgtacagtc aattgactac
     4201 tcacctcttg actgacaaaa attacactac gtacggccac ctgaagaacc agaaaggaca
     4261 ccgcaaatgg tacgttcttg tcaaaaataa ccttcagaat agcgacataa cagctgtgag
     4321 ttcattccgc aacatagttg ctcacatatc cgttgtgagg aatagtaatg aatacataag
     4381 cgggatcgga gaactgcact cttacttcga actttatcac tatctcgttc aatcaatgat
     4441 cgcaaagaat aattggtatg ataccagcca ccagccgaag accgccgaat atctgaacaa
     4501 tctgaaaaag caccacactt attgtaagga ctttgtaaaa gcgtactgta tcccgttcgg
     4561 atacgtcgtg ccgcggtaca aaaacctcac gataaatgaa ctttttgatc gaaataaccc
     4621 gaatcccgaa ccaaaagagg aagtgggatc cggacctaag aaaaagagga aggtggcggc
     4681 cgcttaccca tacgatgttc cagattacgc ttaggctagg aattcaataa aagatcttta
     4741 ttttcattag atctgtgtgt tggttttttg tgtgcggacc gagcggccgc aggaacccct
     4801 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     4861 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     4921 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4981 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5041 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     5101 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     5161 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     5221 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5281 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     5341 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     5401 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     5461 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     5521 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     5581 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     5641 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5701 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5761 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5821 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5881 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5941 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6001 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6061 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6121 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6181 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6241 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6301 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6361 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6421 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6481 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6541 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6601 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6661 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6721 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6781 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6841 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt g
//
