LOCUS       sequence-115341-        1083 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Tribolium U6a promoter with BsaI cloning site to insert guide RNA 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1083)
  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 1083)
  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(U6a-BsaI-gRNA)
FEATURES             Location/Qualifiers
     source          1..1083
                     /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        complement(560..635)
                     /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 gataaaaaag caccgactcg gtgccacttt ttcaagttga taacggacta
      601 gccttatttt aacttgctat ttctagctct aaaactgaga ccctctctcg gtctctcact
      661 tgcccttctc aacaatctgt atatgaacgt agcgacgggc gaccagcgtt agtgagatgt
      721 aacctccaca ataacttttt taattttaga atatgactgg tttttatcaa tttgtttgtg
      781 tttactattt aataagaagt tgttacatta ttagcattaa acttaaaatt atctcaatta
      841 atattaacac ataaataaat aaataaaaca gttagtcatt gatttcgtaa aaggtttagt
      901 ctagactatc cataatcaat aatcatgtac aaattattaa ttgttgatga tacagttctc
      961 tacacgatta atttgataaa gaccataaat tatttatttt tggattacga gaacgtgtta
     1021 acggacgtaa acgaattgtt tttgcaaggg atccttgtaa tggcgcgcca cttaatcccg
     1081 gga
//
