LOCUS       sequence-115343-        1044 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Tribolium U6b promoter with BsaI cloning site to insert guide RNA 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1044)
  AUTHORS   Gilles AF, Schinko JB, Averof M
  TITLE     Efficient CRISPR-mediated gene targeting and transgene replacement 
            in the beetle Tribolium castaneum.
  JOURNAL   Development. 2015 Jul 9. pii: dev.125054.
  PUBMED    26160901
REFERENCE   2  (bases 1 to 1044)
  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: 
            "Development. 2015 Jul 9. pii: dev.125054."
COMMENT     Sequence Label: p(U6b-BsaI-gRNA)
FEATURES             Location/Qualifiers
     source          1..1044
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     216..233
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    350..371
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(386..391,392..409,410..416)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    424..440
                     /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     429..451
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     448..464
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     448..464
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     483..503
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        485..503
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        951..1026
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
       61 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      121 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      181 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      241 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
      301 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
      361 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
      421 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
      481 gcgcaattaa ccctcactaa agggaacaaa agctggagct tggaattcca cgtgatactt
      541 atggccggcc gataaggatc cgggtgttat tattattact tgtgacctgg tgataagtgc
      601 ttcaagtact agagattgag tcactgttca aggattattt aaaaacacaa cacgcagatt
      661 tatcagttac cttatgaaca aatgaatatc tatttttgtt tcaaaactgt tttcattacg
      721 acaaattaat atttcttcaa ctaaataata aaaattttaa ataagcgcca ttacatacaa
      781 atatacgaat cattacgaac gtttgtgttt tgcgatcgat tgtaaccgac caatcaaaat
      841 aaagcgggaa atttcaaatc tcttatatct cactactacg tatcggttgc cgctaactac
      901 atatacaagg caggtagaag caccgttcga gagaccgaga gagggtctca gttttagagc
      961 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
     1021 cggtgctttt ttgtaatggc gcgc
//
