LOCUS       sequence-212870-        1448 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Express a cloned sgRNA from AtU626 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1448)
  AUTHORS   Lawrenson T, Hundleby P, Harwood W
  TITLE     Creating Targeted Gene Knockouts in Brassica oleracea Using 
            CRISPR/Cas9.
  JOURNAL   Methods Mol Biol. 2019;1917:155-170. doi: 
            10.1007/978-1-4939-8991-1_12.
  PUBMED    30610635
REFERENCE   2  (bases 1 to 1448)
  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: "Methods Mol
            Biol."; date: "2019"; volume: "1917:155-170. doi"; pages: " 
            10.1007/978-1-4939-8991-1_12"
COMMENT     Sequence Label: pBoL1P4GA
FEATURES             Location/Qualifiers
     source          1..1448
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    331..352
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(367..372,373..390,391..397)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    405..421
                     /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)."
     RBS             439..450
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             457..1134
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1179..1250
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1266..1293
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    complement(1390..1414)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 cactctgtgg aagacaatta ccatcttcat tcttaagata tgaagataat cttcaaaagg
       61 cccctgggaa tctgaaagaa gagaagcagg cccatttata tgggaaagaa caatagtatt
      121 tcttatatag gcccatttaa gttgaaaaca atcttcaaaa gtcccacatc gcttagataa
      181 gaaaacgaag ctgagtttat atacagctag agtcgaagta gtgcgagacg caatacgcaa
      241 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
      301 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
      361 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
      421 atttcacaca tactagagaa agaggagaaa tactagatgg cttcctccga ggacgttatc
      481 aaagagttca tgcgtttcaa agttcgtatg gaaggttccg ttaacggtca cgagttcgaa
      541 atcgaaggtg aaggtgaagg tcgtccgtac gaaggtaccc agaccgctaa actgaaagtt
      601 accaaaggtg gtccgctgcc gttcgcttgg gacatcctgt ccccgcagtt ccagtacggt
      661 tccaaagctt acgttaaaca cccggctgac atcccggact acctgaaact gtccttcccg
      721 gaaggtttca aatgggaacg tgttatgaac ttcgaggacg gtggtgttgt taccgttacc
      781 caggactcct ccctgcaaga cggtgagttc atctacaaag ttaaactgcg tggtaccaac
      841 ttcccgtccg acggtccggt tatgcagaaa aaaaccatgg gttgggaagc ttccaccgaa
      901 cgtatgtacc cggaggacgg tgctctgaaa ggtgaaatca aaatgcgtct gaaactgaaa
      961 gacggtggtc actacgacgc tgaagttaaa accacctaca tggctaaaaa accggttcag
     1021 ctgccgggtg cttacaaaac cgacatcaaa ctggacatca cctcccacaa cgaggactac
     1081 accatcgttg aacagtacga acgtgctgaa ggtcgtcact ccaccggtgc ttaataacgc
     1141 tgatagtgct agtgtagatc gctactagag ccaggcatca aataaaacga aaggctcagt
     1201 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc
     1261 acactggctc accttcgggt gggcctttct gcgtttatac gtctcttaga gctagaaata
     1321 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     1381 tttttctaga cccagctttc ttgtacaaag ttggcattac gctttacggg aaagtcttcc
     1441 acgaagtg
//
