LOCUS       sequence-419129-        2288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human expression plasmid for SaCas9 sgRNA(84nt in length) (need to 
            clone in spacer into BsmBI sites): U6-BsmBIcassette-Sa-sgRNA(84).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2288)
  AUTHORS   Kleinstiver BP, Prew MS, Tsai SQ, Nguyen NT, Topkar VV, Zheng Z, 
            Joung JK
  TITLE     Broadening the targeting range of Staphylococcus aureus CRISPR-Cas9 
            by modifying PAM recognition.
  JOURNAL   Nat Biotechnol. 2015 Nov 2. doi: 10.1038/nbt.3404.
  PUBMED    26524662
REFERENCE   2  (bases 1 to 2288)
  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 
            Biotechnol. 2015 Nov 2. doi: 10.1038/nbt.3404."
COMMENT     Sequence Label: BPK2660
FEATURES             Location/Qualifiers
     source          1..2288
                     /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(539..1127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(619..638)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1298..2089,2090..2158))
                     /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     1921..1940
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2159..2263)
                     /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 cgagacgatt aatgcgtctc ggttttagta
      361 ctctgtaatg aaaattacag aatctactaa aacaaggcaa aatgccgtgt ttatctcgtc
      421 aacttgttgg cgagattttt ttaagcttgg gccgctcgag gtacctctct acatatgaca
      481 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
      541 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
      601 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
      661 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
      721 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
      781 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
      841 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
      901 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
      961 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     1021 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1081 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1141 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     1201 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     1261 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     1321 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     1381 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     1441 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     1501 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     1561 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     1621 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     1681 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     1741 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     1801 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     1861 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     1921 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     1981 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     2041 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     2101 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     2161 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     2221 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     2281 tgccacct
//
