LOCUS       sequence-268588-        2284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  U6 promoter sgRNA entry vector used for all St1Cas9 sgRNAs (clone 
            spacer oligos into BsmBI cassette) - similar to V1 sgRNA 
            architecture from Carter et al. biorxiv 2018.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2284)
  AUTHORS   Mahendra C, Christie KA, Osuna BA, Pinilla-Redondo R, Kleinstiver 
            BP, Bondy-Denomy J
  TITLE     Broad-spectrum anti-CRISPR proteins facilitate horizontal gene 
            transfer.
  JOURNAL   Nat Microbiol. 2020 Apr;5(4):620-629. doi: 
            10.1038/s41564-020-0692-2. Epub 2020 Mar 26.
  PUBMED    32218510
REFERENCE   2  (bases 1 to 2284)
  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"; doi: 
            "10.1038/s41564-020-0692-2"; journalName: "Nat Microbiol"; date: 
            "2020-04"; volume: "5"; issue: "4"; pages: "620-629"
COMMENT     Sequence Label: pUC19-U6-BsmBI_cassette-St1Cas9-sgRNA (KAC14)
FEATURES             Location/Qualifiers
     source          1..2284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        complement(794..1042)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(852..871)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1022..1042)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        1149..1253
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1254..1322,1323..2114)
                     /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(1472..1491)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt catatgtaga
      661 gaggtacctc gagcggccca agcttaaaaa aacaccctgc cataaaatga cagggtgttg
      721 atttcggcat gaagccttat ctttgtagct tctggtacca gagtacaaag accgagacgc
      781 attaatcgtc tcgggtgttt cgtcctttcc acaagatata taaagccaag aaatcgaaat
      841 actttcaagt tacggtaagc atatgatagt ccattttaaa acataatttt aaaactgcaa
      901 actacccaag aaattattac tttctacgtc acgtattttg tactaatatc tttgtgttta
      961 cagtcaaatt aattctaatt atctctctaa cagccttgta tcgtatatgc aaatatgaag
     1021 gaatcatggg aaataggccc tcttcctgcc cgaccttggt accggatcca gtcgactgaa
     1081 ttggttcctt taaagcctgc ttttttgtac agctagcgac gtcaggtggc acttttcggg
     1141 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1201 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1261 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1321 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1381 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1441 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     1501 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1561 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1621 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1681 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1741 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1801 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1861 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1921 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1981 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     2041 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     2101 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     2161 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2221 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2281 cttc
//
