LOCUS       sequence-477973-        6428 bp    DNA     circular SYN 09-JUL-2025
DEFINITION  Expression of guide RNA for PspCas13b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6428)
  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 6428)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 9, 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-Control
FEATURES             Location/Qualifiers
     source          1..6428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3311..3899)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3391..3410)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4070..4861,4862..4930))
                     /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     4693..4712
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4931..5035)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5103..5121
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5159..5181)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5281..5300
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5317..5772)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5454..5475)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5666..5685
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        complement(5908..6156)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(5966..5985)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(6136..6156)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 aaactctcac tcttaccgaa cttggccgta acctgccaac caaagcgaga acaaaacata
       61 acatcaaacg aatcgaccga ttgttaggta atcgtcacct ccacaaagag cgactcgctg
      121 tataccgttg gcatgctagc tttatctgtt cgggcaatac gatgcccatt gtacttgttg
      181 actggtctga tatccgtgag caaaaacggc ttatggtatt gcgagcttca gtcgcactac
      241 acggtcgttc tgttactctt tatgagaaag cgttcccgct ttcagagcaa tgttcaaaga
      301 aagctcatga ccaatttcta gccgaccttg cgagcattct accgagtaac accacaccgc
      361 tcattgtcag tgatgctggc tttaaagtgc catggtataa atccgttgag aagctgggtt
      421 ggtactggtt aagtcgagta agaggaaaag tacaatatgc agacctagga gcggaaaact
      481 ggaaacctat cagcaactta catgatatgt catctagtca ctcaaagact ttaggctata
      541 agaggctgac taaaagcaac ccaatctcat gccaaattct attgtataaa tctcgctcta
      601 aaggccgaaa aaatcagcgc tcgacacgga ctcattgtca ccacccgtca cctaaaatct
      661 actcagcgtc ggcaaaggag ccatggattc tagcaactaa cttacctgtt gaaattcgaa
      721 cacccaaaca acttgttaat atctattcga agcgaatgca gattgaagaa accttccgag
      781 acttgaaaag tcctgcctac ggactaggcc tacgccatag ccgaacgagc agctcagagc
      841 gttttgatat catgctgcta atcgccctga tgcttcaact aacatgttgg cttgcgggcg
      901 ttcatgctca gaaacaaggt tgggacaagc acttccaggc taacacagtc agaaatcgaa
      961 acgtactctc aacagttcgc ttaggcatgg aagttttgcg gcattctggc tacacaataa
     1021 caagggaaga ctcactcgtg gctgcaaccc tgcttactca aaatctattc acacatggtt
     1081 acgttttggg gaaattatga ggggatctct cagcaccctg cgtgagtttc acgagaagct
     1141 gcgtgaactg ctgacgacgc tgaatctgaa gctggaaccg gcggatgatt ttcgtgacga
     1201 gccggtgaag ttaacggcgt gagtgaaatg tgccggatgc atcacatccg gcaatattca
     1261 ttaaaactga tacgtcatgc caaccgcgac aatatcatca ttattaatat tcaatttgtt
     1321 atcgctatcc agttggttga ttttataatc aacaaacgct gacatatttt tgttgaaata
     1381 atacgtagca ccgacgtcga tataattgac cagatcttca tcaccgatac cttcaatatc
     1441 tttccctttc gataagacat aacccagcga tggacgcaga ccaaagtcaa actggtattg
     1501 agcgaccgct tcaaagttct gtgtcttatt ggcaaagccg ccagttattg gcgtcatttt
     1561 gcgtgtttca gaatagaaag ttgccagata aatattattg gcatcgtatt tcagacctgt
     1621 tgcccatgct tctgcacgct tgcctgtgcc acggctttgc aggttctgct cgttggtgcg
     1681 atctgagttg gtataggccc cactaatggc gaaatcgctg ccgccaaagt catatgtcaa
     1741 tgacgtgccg aagccatcgc cgttttgctt tttaacgtcg cggttttcgt ttttcccttg
     1801 atattgcagg gttaagttca ggccatcgat aacgccgaag aagtcggtgt tccgatacgt
     1861 cgccagaccg ctggcgcgtt tggtcataaa gttgtcggtc tgcgccgagg aatcgccacc
     1921 aaattccggg aacatatcgg tccaggcttc cacgtcatac aacgccctga tgaatcccct
     1981 aatgattttg gtaaaaatca ttaagttaag gtggatacac atcttgtcat atgatcaaat
     2041 ggtttcgcga aaaatcaata atcagacaac aagatgtgcg aactcgatat tttacacgac
     2101 tctctttacc aattctgccc cgaattacac ttaaaacgac tcaacagctt aacgttggct
     2161 tgccacgcat tacttgactg taaaactctc actcttaccg aacttggccg taacctgcca
     2221 accaaagcga gaacaaaaca taacatcaaa cgaatcgacc gattgttagg taatcgtcac
     2281 ctccacaaag agcgactcgc tgtataccgt tggcatgcta gctttatctg ttcgggcaat
     2341 acgatgccca ttgtacttgt tgactggtct gatattcgtg agcaaaaacg acttatggta
     2401 ttgcgagctt cagtcgcact acacggtcgt tctgttactc tttatgagaa agcgttcccg
     2461 ctttcagagc aatgttcaaa gaaagctcat gaccaatttc tagccgacct tgcgagcatt
     2521 ctaccgagta acaccacacc gctcattgtc agtgatgctg gctttaaagt gccatggtat
     2581 aaatccgttg agaagctggg ttggtactgg ttaagtcgag taagaggaaa agtacaatat
     2641 gcagacctag gagcggaaaa ctggaaacct atcagcaact tacatgatat gtcatctagt
     2701 cactcaaaga ctttaggcta taagaggctg actaaaagca atccaatctc atgccaaatt
     2761 ctattgtata aatctcgctc taaaggccga aaaaatcagc gctcgacacg gactcattgt
     2821 caccacccgt cacctaaaat ctactcagcg tcggcaaagg agccatgggt tctagcaact
     2881 aacttacctg ttgaaattcg aacacccaaa caacttgtta atatctattc gaagcgaatg
     2941 cagattgaag aaaccttccg agacttgaaa agtcctgcct acggactagg cctacgccat
     3001 agccgaacga gcagctcaga gcgttttgat atcatgctgc taatcgccct gatgcttcaa
     3061 ctaacatgtt ggcttgcggg cgttcatgct cagaaacaag gttgggacaa gcacttccag
     3121 gctaacacag tcagaaatcg aaacgtactc tcaacagttc gcttaggcat ggaagttttg
     3181 cggcattctg gctacacaat aacaagggaa gacttactcg tggctgcaac cctactagct
     3241 caaaatttat tcacacatgg ttacgctttg gggaaattat gaggggatct ctcagcgcgt
     3301 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3361 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3421 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3481 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3541 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3601 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3661 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3721 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3781 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3841 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3901 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3961 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4021 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4081 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4141 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4201 tgataccgcg gcttccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4261 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4321 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4381 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4441 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4501 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4561 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4621 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4681 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4741 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4801 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4861 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4921 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4981 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5041 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     5101 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     5161 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     5221 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     5281 cggcatcaga gcagattgta ctgagagtgc accataaaat tgtaaacgtt aatattttgt
     5341 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag accgaaatcg
     5401 gcaaaatccc ttataaatca aaagaatagc ccgagataga gttgagtgtt gttccagttt
     5461 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     5521 atcagggcga tggcccacta cgtgaaccat cacccaaatc aagttttttg gggtcgaggt
     5581 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     5641 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctaaggcgc
     5701 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     5761 tacagggcgc gtactatggt tgctttgacg tatgcggtgt gaaataccgc acagatgcgt
     5821 aaggagaaaa taccgcatca gaattcaaaa aaaagttgta atagcccctc aaaactggac
     5881 cttccacaac ggtcttctcg aagacccggt gtttcgtcct ttccacaaga tatataaagc
     5941 caagaaatcg aaatactttc aagttacggt aagcatatga tagtccattt taaaacataa
     6001 ttttaaaact gcaaactacc caagaaatta ttactttcta cgtcacgtat tttgtactaa
     6061 tatctttgtg tttacagtca aattaattcc aattatctct ctaacagcct tgtatcgtat
     6121 atgcaaatat gaaggaatca tgggaaatag gccctcgcta gctgagcaaa aggccagcaa
     6181 aaggccagga accgtaaaaa ggccgcgttg ctctgatgaa tcccctaatg atttttatca
     6241 aaatcattaa gttaaggtag atacacatct tgtcatatga tcaaatggtt tcgccaaaaa
     6301 tcaataatca gacaacaaaa tgtgcgaact cgatatttta cacgactctc tttaccaatt
     6361 ctgccccgaa ttacacttaa aacgactcaa cagcttaacg ttggcttgcc acgccttact
     6421 tgactgta
//
