LOCUS       sequence-311141-        3127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Binary vector for expression of a gRNA targeting AtAP1 promoter .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3127)
  AUTHORS   Ochoa-Fernandez R, Abel NB, Wieland FG, Schlegel J, Koch LA, Miller 
            JB, Engesser R, Giuriani G, Brandl SM, Timmer J, Weber W, Ott T, 
            Simon R, Zurbriggen MD
  TITLE     Optogenetic control of gene expression in plants in the presence of 
            ambient white light.
  JOURNAL   Nat Methods. 2020 Jul;17(7):717-725. doi: 10.1038/s41592-020-0868-y.
            Epub 2020 Jun 29.
  PUBMED    32601426
REFERENCE   2  (bases 1 to 3127)
  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"; doi: 
            "10.1038/s41592-020-0868-y"; journalName: "Nat Methods"; date: 
            "2020-07"; volume: "17"; issue: "7"; pages: "717-725"
COMMENT     Sequence Label: pROF441
FEATURES             Location/Qualifiers
     source          1..3127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..23
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     misc_RNA        453..528
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(540..564)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    586..610
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(701..1289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(781..800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1460..2275)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1460..2275)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2183..2202
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2267..2286
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2566..3001
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tggcaggata tattgtggtg taacaagctt cgtctcagtc aggagctttt tttcttcttc
       61 ttcgttcata cagttttttt ttgtttatca gcttacattt tcttgaaccg tagctttcgt
      121 tttcttcttt ttaactttcc attcggagtt tttgtatctt gtttcatagt ttgtcccagg
      181 attagaatga ttaggcatcg aaccttcaag aatttgattg aataaaacat cttcattctt
      241 aagatatgaa gataatcttc aaaaggcccc tgggaatctg aaagaagaga agcaggccca
      301 tttatatggg aaagaacaat agtatttctt atataggccc atttaagttg aaaacaatct
      361 tcaaaagtcc cacatcgctt agataagaaa acgaagctga gtttatatac agctagagtc
      421 gaagtagtga ttgtatatct cgtactaatg tcgttttaga gctagaaata gcaagttaaa
      481 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttctaga
      541 cccagctttc ttgtacaaag ttggcattac gcttgagacg aagcttgaca ggatatattg
      601 gcgggtaaac taagtcgctg tatgtgtttg tttgagatct catgtgagca aaaggccagc
      661 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
      721 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
      781 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
      841 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
      901 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
      961 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt tagtccaacc
     1021 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     1081 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1141 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaga agagttggta
     1201 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     1261 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     1321 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     1381 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     1441 tgtaacattg gtctagtgat tagaaaaact catcgagcat caaatgaaac tgcaatttat
     1501 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     1561 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     1621 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     1681 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
     1741 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     1801 cgttattcat tcgtgattgc gcctgagcga gtcgaaatac gcgatcgctg ttaaaaggac
     1861 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     1921 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttccct gggatcgcag
     1981 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     2041 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     2101 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cggtagattg
     2161 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     2221 tgttggaatt taatcgcggc cttgagcaag acgtttcccg ttgaatatgg ctcataacac
     2281 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat
     2341 cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttgttga ataaatcgaa
     2401 cttttgctga gttgaaggat cagatcacgc atcttcccga caacgcagac cgttccgtgg
     2461 caaagcaaaa gttcaaaatc accaactggt ccacctacaa caaagctctc atcaaccgtg
     2521 gctccctcac tttctggctg gatgatgggg cgattcaggc gatccccatc caacagcccg
     2581 ccgtcgagcg ggctttttta tccccggaag cctgtggata gagggtagtt atccacgtga
     2641 aaccgctaat gccccgcaaa gccttgattc acggggcttt ccggcccgct ccaaaaacta
     2701 tccacgtgaa atcgctaatc agggtacgtg aaatcgctaa tcggagtacg tgaaatcgct
     2761 aataaggtca cgtgaaatcg ctaatcaaaa aggcacgtga gaacgctaat agccctttca
     2821 gatcaacagc ttgcaaacac ccctcgctcc ggcaagtagt tacagcaagt agtatgttca
     2881 attagctttt caattatgaa tatatatatc aattattggt cgcccttggc ttgtggacaa
     2941 tgcgctacgc gcaccggctc cgcccgtgga caaccgcaag cggttgccca ccgtcgagcg
     3001 cctttgccca caacccggcg gccggccgca acagatcgtt ttataaattt ttttttttga
     3061 aaaagaaaaa gcccgaaagg cggcaacctc tcgggcttct ggatttccga tccccggaat
     3121 tagatct
//
