LOCUS       sequence-391389-        2290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression construct encoding a S. aureus guide RNA targeting the 
            human CLYBL safe harbor locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2290)
  AUTHORS   Wang Q, Liu J, Janssen JM, Goncalves MAFV
  TITLE     Precise homology-directed installation of large genomic edits in 
            human cells with cleaving and nicking high-specificity Cas9 
            variants.
  JOURNAL   Nucleic Acids Res. 2023 Mar 17:gkad165. doi: 10.1093/nar/gkad165.
  PUBMED    36928106
REFERENCE   2  (bases 1 to 2290)
  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. 2023 Mar 17:gkad165. doi: 10.1093/nar/gkad165."
COMMENT     Sequence Label: AM77_pU6.Sa-gRNA.CLYBL
FEATURES             Location/Qualifiers
     source          1..2290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        82..330
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     82..102
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     253..272
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(541..1129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(621..640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1300..2091,2092..2160))
                     /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     1923..1942
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2161..2265)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc
       61 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata
      121 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta
      181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
      241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
      301 ttatatatct tgtggaaagg acgaaacacc gcctacatac cgttatcctg tgggttttag
      361 tactctgtaa tgaaaattac agaatctact aaaacaaggc aaaatgccgt gtttatctcg
      421 tcaacttgtt ggcgagattt ttttaagctt gggccgctcg aggtacctct ctacatatga
      481 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
      541 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
      601 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      661 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      721 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      781 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      841 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      901 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      961 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     1021 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1081 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     1141 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     1201 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     1261 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     1321 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     1381 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     1441 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     1501 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1561 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     1621 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1681 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1741 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1801 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1861 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1921 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1981 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     2041 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     2101 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     2161 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     2221 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     2281 agtgccacct
//
