LOCUS       sequence-478755-        4259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of guide RNA targeting the start codon of the Renilla 
            luciferase gene for PspCas13b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4259)
  AUTHORS   Apostolopoulos A, Kawamoto N, Chow SYA, Tsuiji H, Ikeuchi Y, 
            Shichino Y, Iwasaki S
  TITLE     dCas13-mediated translational repression for accurate gene silencing
            in mammalian cells.
  JOURNAL   Nat Commun. 2024 Mar 11;15(1):2205. doi: 10.1038/s41467-024-46412-7.
  PUBMED    38467613
REFERENCE   2  (bases 1 to 4259)
  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: "2024-03-11"; volume: "15(1):2205. doi"; pages: " 
            10.1038/s41467-024-46412-7"
COMMENT     Sequence Label: pC016-gPsp-AUG16_start_codon
FEATURES             Location/Qualifiers
     source          1..4259
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      138..726
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     627..646
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2127..2375
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2127..2147
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2298..2317
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     rep_origin      2524..2979
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2611..2630)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2821..2842
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2996..3015)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3115..3137
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3175..3193)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3261..3365
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3366..3434,3435..4226)
                     /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(3584..3603)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
       61 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
      121 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
      181 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
      241 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
      301 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
      361 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
      421 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
      481 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
      541 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
      601 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
      661 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
      721 tggaaaaacg ccagcaacgc gctgagagat cccctcataa tttccccaaa acgtaaccat
      781 gtgtgaatag attttgagta agcagggttg cagccacgag tgagtcttcc cttgttattg
      841 tgtagccaga atgccgcaaa acttccatgc ctaagcgaac tgttgagagt acgtttcgat
      901 ttctgactgt gttagcctgg aagtgcttgt cccaaccttg tttctgagca tgaacgcccg
      961 caagccaaca tgttagttga agcatcaggg cgattagcag catgatatca aaacgctctg
     1021 agctgctcgt tcggctatgg cgtaggccta gtccgtaggc aggacttttc aagtctcgga
     1081 aggtttcttc aatctgcatt cgcttcgaat agatattaac aagttgtttg ggtgttcgaa
     1141 tttcaacagg taagttagtt gctagaatcc atggctcctt tgccgacgct gagtagattt
     1201 taggtgacgg gtggtgacaa tgagtccgtg tcgagcgctg attttttcgg cctttagagc
     1261 gagatttata caatagaatt tggcatgaga ttgggttgct tttagtcagc ctcttatagc
     1321 ctaaagtctt tgagtgacta gatgacatat catgtaagtt gctgataggt ttccagtttt
     1381 ccgctcctag gtctgcatat tgtacttttc ctcttactcg acttaaccag taccaaccca
     1441 gcttctcaac ggatttatac catggcactt taaagccagc atcactgaca atgagcggtg
     1501 tggtgttact cggtagaatg ctcgcaaggt cggctagaaa ttggtcatga gctttctttg
     1561 aacattgctc tgaaagcggg aacgctttct cataaagagt aacagaacga ccgtgtagtg
     1621 cgactgaagc tcgcaatacc ataagccgtt tttgctcacg gatatcagac cagtcaacaa
     1681 gtacaatggg catcgtattg cccgaacaga taaagctagc atgccaacgg tatacagcga
     1741 gtcgctcttt gtggaggtga cgattaccta acaatcggtc gattcgtttg atgttatgtt
     1801 ttgttctcgc tttggttggc aggttacggc caagttcggt aagagtgaga gttttacagt
     1861 caagtaaggc gtggcaagcc aacgttaagc tgttgagtcg ttttaagtgt aattcggggc
     1921 agaattggta aagagagtcg tgtaaaatat cgagttcgca cattttgttg tctgattatt
     1981 gatttttggc gaaaccattt gatcatatga caagatgtgt atctacctta acttaatgat
     2041 tttgataaaa atcattaggg gattcatcag agcaacgcgg cctttttacg gttcctggcc
     2101 ttttgctggc cttttgctca gctagcgagg gcctatttcc catgattcct tcatatttgc
     2161 atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga
     2221 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
     2281 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
     2341 tggctttata tatcttgtgg aaaggacgaa acaccgtaca ccttggaagc cattgtgaac
     2401 ggggcgttgt ggaaggtcca gttttgaggg gctattacaa cttttttttg aattctgatg
     2461 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     2521 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2581 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2641 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2701 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     2761 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2821 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     2881 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     2941 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     3001 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     3061 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3121 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     3181 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     3241 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3301 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3361 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3421 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3481 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3541 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3601 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3661 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3721 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3781 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3841 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3901 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3961 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4021 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4081 gaagccgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4141 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4201 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttag
//
