LOCUS       sequence-357068-        2990 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PasCas13b-gRNA targeting Nluc(W12stop) for RNA editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2990)
  AUTHORS   Somiya M, Kuroda S
  TITLE     Verification of extracellular vesicle-mediated functional mRNA 
            delivery by RNA editing
  JOURNAL   bioRxiv 2022.01.25.477620
REFERENCE   2  (bases 1 to 2990)
  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: "bioRxiv 
            2022.01.25.477620"
COMMENT     Sequence Label: PasCas13b-gRNA-Nluc(W12stop)
FEATURES             Location/Qualifiers
     source          1..2990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     150..167
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    284..305
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(320..325,326..343,344..350)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    358..374
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     382..398
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     382..398
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        412..660
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     412..432
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     583..602
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(772..789)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(772..788)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(781..803)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(997..1016)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1116..1138
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1176..1194)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1262..1366
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1367..1435,1436..2227)
                     /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(1585..1604)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2398..2986
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2887..2906
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
       61 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
      121 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
      181 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
      241 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
      301 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
      361 tgagcggata acaatttcac gcaggaaaca gctatgacca tgattacgcc agagggccta
      421 tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga gataattgga
      481 attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa
      541 tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc
      601 gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc
      661 agccggctgt ctgtcgccag tccccaacga aatcttcgag tgtgaagacc agttgtggaa
      721 ggtccagttt tgaggggcta ttacaacttt tttggtaccg agctcgaatt cactggccgt
      781 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
      841 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
      901 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
      961 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     1021 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     1081 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     1141 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     1201 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     1261 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1321 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1381 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1441 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1501 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1561 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     1621 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     1681 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     1741 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1801 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1861 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1921 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1981 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2041 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     2101 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     2161 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2221 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     2281 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     2341 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2401 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2461 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2521 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     2581 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2641 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2701 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2761 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2821 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2881 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2941 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
//
