LOCUS       sequence-453804-        1583 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cloning vector to clone sgRNA in scaffold under TaU6 promoter for 
            Cas9 mediated CRISPR for goldengate level1 position 4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1583)
  AUTHORS   Winkelmolen T, Colleoni P, Moscou M, Hoseinzadeh P, Oldach K, 
            Schmidt RC, Immink R, van Esse GW
  TITLE     VRS5 (HvTB1) binds to the promoter of tillering and floral homeotic 
            genes to regulate their expression
  JOURNAL   bioRxiv 2024.08.22.609225
REFERENCE   2  (bases 1 to 1583)
  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: "bioRxiv 
            2024.08.22.609225"
COMMENT     Sequence Label: L1p4_barley_sgRNA_scaffold
FEATURES             Location/Qualifiers
     source          1..1583
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    477..498
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(513..518,519..536,537..543)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    551..567
                     /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             585..596
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             603..1280
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1325..1396
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1412..1439
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    complement(1550..1574)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ggagtagacc aagcccgtta ttctgacagt tctggtgctc aacacattta tatttatcaa
       61 ggagcacatt gttactcact gctaggaggg aatcgaacta ggaatattga tcagaggaac
      121 tacgagagag ctgaagataa ctgccctcta gctctcactg atctgggtcg catagtgaga
      181 tgcagcccac gtgagttcag caacggtcta gcgctgggct tttaggcccg catgatcggg
      241 cttttgtcgg gtggtcgacg tgttcacgat tggggagagc aacgcagcag ttcctcttag
      301 tttagtccca cctcgcctgt ccagcagagt tctgaccggt ttataaactc gcttgctgca
      361 tcagacttgc gagacgcaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
      421 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
      481 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
      541 ttgtgtggaa ttgtgagcgg ataacaattt cacacatact agagaaagag gagaaatact
      601 agatggcttc ctccgaggac gttatcaaag agttcatgcg tttcaaagtt cgtatggaag
      661 gttccgttaa cggtcacgag ttcgaaatcg aaggtgaagg tgaaggtcgt ccgtacgaag
      721 gtacccagac cgctaaactg aaagttacca aaggtggtcc gctgccgttc gcttgggaca
      781 tcctgtcccc gcagttccag tacggttcca aagcttacgt taaacacccg gctgacatcc
      841 cggactacct gaaactgtcc ttcccggaag gtttcaaatg ggaacgtgtt atgaacttcg
      901 aggacggtgg tgttgttacc gttacccagg actcctccct gcaagacggt gagttcatct
      961 acaaagttaa actgcgtggt accaacttcc cgtccgacgg tccggttatg cagaaaaaaa
     1021 ccatgggttg ggaagcttcc accgaacgta tgtacccgga ggacggtgct ctgaaaggtg
     1081 aaatcaaaat gcgtctgaaa ctgaaagacg gtggtcacta cgacgctgaa gttaaaacca
     1141 cctacatggc taaaaaaccg gttcagctgc cgggtgctta caaaaccgac atcaaactgg
     1201 acatcacctc ccacaacgag gactacacca tcgttgaaca gtacgaacgt gctgaaggtc
     1261 gtcactccac cggtgcttaa taacgctgat agtgctagtg tagatcgcta ctagagccag
     1321 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     1381 gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt
     1441 ttatacgtct ccgtttcaga gctatgctgg aaacagcata gcaagttgaa ataaggctag
     1501 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttctaga cccagctttc
     1561 ttgtacaaag ttggcattac gct
//
