LOCUS       sequence-361296-        2217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constituitvely expressed CasRx crRNA cloning vector, truncated 3' 
            terminator region.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2217)
  AUTHORS   Otoupal PB, Cress BF, Doudna JA, Schoeniger JS
  TITLE     CRISPR-RNAa: targeted activation of translation using dCas13 fusions
            to translation initiation factors.
  JOURNAL   Nucleic Acids Res. 2022 Aug 11. pii: 6660959. doi: 
            10.1093/nar/gkac680.
  PUBMED    35950485
REFERENCE   2  (bases 1 to 2217)
  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: "Nucleic 
            Acids Res. 2022 Aug 11. pii: 6660959. doi: 10.1093/nar/gkac680."
COMMENT     Sequence Label: pPBO.ACT005
FEATURES             Location/Qualifiers
     source          1..2217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        28..62
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        63..92
                     /label=RfxCas13d DR30
                     /note="5' processed 30-base direct repeat for an RfxCas13d 
                     gRNA (Konermann et al., 2018)"
     terminator      109..155
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(208..225)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(379..967)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(459..478)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1138..1929,1930..1998))
                     /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     1761..1780
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1999..2103)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2171..2189
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(join(2193..2217,1..19))
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 tttcgctaag gatgatttct ggaattcttg acagctagct cagtcctagg tataatacta
       61 gtaaccccta ccaactggtc ggggtttgaa acggagacga acgtctcaca aataaaacga
      121 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaactggatc
      181 cttactcgag tctagactgc aggcttcctc gctcactgac tcgctgcgct cggtcgttcg
      241 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      301 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      361 ggccgcgttg ctggcgtttt tccacaggct ccgcccccct gacgagcatc acaaaaatcg
      421 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      481 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      541 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      601 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      661 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      721 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      781 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
      841 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
      901 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
      961 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1021 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1081 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1141 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1201 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1261 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1321 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1381 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1441 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1501 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1561 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1621 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1681 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1741 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1801 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1861 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1921 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     1981 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2041 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2101 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     2161 aacctataaa aataggcgta tcacgaggca gaatttcaga taaaaaaaat ccttagc
//
