LOCUS       sequence-119950-        1139 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Empty vector for cloning sgRNA. NotI and BamHI for cloning sgRNA. 
            PU6 driven sgRNA. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1139)
  AUTHORS   Norris AD, Kim HM, Colaiacovo MP, Calarco JA
  TITLE     Efficient Genome Editing in Caenorhabditis elegans with a Toolkit of
            Dual-Marker Selection Cassettes.
  JOURNAL   Genetics. 2015 Oct;201(2):449-58. doi: 10.1534/genetics.115.180679. 
            Epub 2015 Jul 30.
  PUBMED    26232410
REFERENCE   2  (bases 1 to 1139)
  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: 
            "10.1534/genetics.115.180679"; journalName: "Genetics"; date: 
            "2015-10"; volume: "201"; issue: "2"; pages: "449-58"
COMMENT     Sequence Label: pHKMC1: Empty sgRNA for Cloning
FEATURES             Location/Qualifiers
     source          1..1139
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     219..236
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    353..374
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(389..394,395..412,413..419)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    427..443
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     432..454
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     451..467
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     451..467
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_feature    486..722
                     /label=3' CeU6
                     /note="region downstream of the C. elegans K09B11.12 U6 
                     snRNA gene"
     misc_RNA        complement(730..805)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
       61 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      121 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      181 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      241 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
      301 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
      361 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
      421 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
      481 agcttcacag ccgactatgt ttggcgtcga gattttgtaa ttttctttgg aatattgtca
      541 aaaaaaatca attttttttc gagtttccta ataaaatttt ttggaccaat ttaatcaact
      601 ctggcaactt taaaatcgtc cctccgggaa tttaccttta accaagatta gttgaaacat
      661 tgaactcaaa aatttagagt atttcatttt agtttcaaaa agggatatct agaatattat
      721 tgaaaaaaag caccgactcg gtgccacttt ttcaagttga taacggacta gccttatttt
      781 aacttgctat ttctagctct aaaacatgga tccggtaagc ggccgcaaac atttagattt
      841 gcaattcaat tatataggga ccatttatca tgcagctgac atattgagag tcctgtaaga
      901 cactatacca aatggggatg tagatcaata atttgtgggg gaggcatgtg tgattcgtac
      961 tcgttcggat agagctttgg tgaatgtcca ggcttctgaa acgtaggtga gcgctggaag
     1021 gacaattgag tcgaacagat tcgcacgaat cttcttgtcg gtgatggagt cggtggcatt
     1081 cttgattcca ttgaatgcag cccaggctgc tcgacgtctt cggtggattt ccggcatca
//
