LOCUS       sequence-137370-         866 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA scaffold and human U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 866)
  AUTHORS   Vidigal JA, Ventura A
  TITLE     Rapid and efficient one-step generation of paired gRNA CRISPR-Cas9 
            libraries.
  JOURNAL   Nat Commun. 2015 Aug 17;6:8083. doi: 10.1038/ncomms9083.
  PUBMED    26278926
REFERENCE   2  (bases 1 to 866)
  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 
            Commun."; date: "2015-08-17"; volume: "6:8083. doi"; pages: " 
            10.1038/ncomms9083"
COMMENT     Sequence Label: pDonor_hU6
FEATURES             Location/Qualifiers
     source          1..866
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        31..106
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        197..445
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     197..217
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     368..387
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(469..485)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(470..486)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(515..535)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(515..533)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(554..570)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(554..570)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(567..589)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    578..594
                     /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)."
     promoter        complement(join(602..608,609..626,627..632))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    647..668
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(785..802)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccccgggctg caggaattcg atgaagacct gttttagagc tagaaatagc aagttaaaat
       61 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
      121 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
      181 gacaaatggc tctagagagg gcctatttcc catgattcct tcatatttgc atatacgata
      241 caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca
      301 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
      361 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
      421 tatcttgtgg aaaggacgaa acaccgtttg ggtcttcatc aagcttatcg ataccgtcga
      481 cctcgagggg gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct
      541 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
      601 acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
      661 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
      721 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
      781 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
      841 actcaaaggc ggtaatacgg ttatcc
//
