LOCUS       sequence-393505-        3428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MODULE B Csy4 vector with 6x gRNAs targeting the four MsPho2-1abcd 
            and MsPho2-2abcd genes in alfalfa (pMOD_B2103 - #91061).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3428)
  AUTHORS   Miller SS, Dornbusch MR, Farmer AD, Huertas R, Gutierrez-Gonzalez 
            JJ, Young ND, Samac DA, Curtin SJ
  TITLE     Alfalfa (Medicago sativa L.) pho2 mutant plants hyperaccumulate 
            phosphate.
  JOURNAL   G3 (Bethesda). 2022 May 30;12(6). pii: 6574359. doi: 
            10.1093/g3journal/jkac096.
  PUBMED    35471600
REFERENCE   2  (bases 1 to 3428)
  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: "G3 
            (Bethesda)."; date: "2022-05-30"; volume: "12(6). pii: 6574359. 
            doi"; pages: " 10.1093/g3journal/jkac096"
COMMENT     Sequence Label: pMOD_B-Pho2-1/2-2-Csy4
FEATURES             Location/Qualifiers
     source          1..3428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      80..668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     569..588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     822..839
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1350..1369
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        1390..1465
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    1466..1485
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        1506..1581
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    1582..1601
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        1622..1697
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    1698..1717
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        1738..1813
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    1814..1833
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        1854..1929
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    1930..1949
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        1970..2045
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    2046..2065
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     polyA_signal    2085..2259
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        2336..2440
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccttgacgg caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
       61 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
      121 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ctaactcttt
      181 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      241 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      301 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      361 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgctcacagc
      421 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      481 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      541 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      601 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      661 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      721 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
      781 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
      841 aagcgggaga gcgcccacct gccaggggac ggcgcgctgg cagacatact gtcccacaaa
      901 tgaagatgga atctgtaaaa gaaaacgcgt gaaataatgc gtctgacaaa ggttaggtcg
      961 gctgccttta atcaatacca aagtggtccc taccacgatg gaaaaactgt gcagtcggtt
     1021 tggctttttc tgacgaacaa ataagattcg tggccgacag gtgggggtcc accatgtgaa
     1081 ggcatcttca gactccaata atggagcaat gacgtaaggg cttacgaaat aagtaagggt
     1141 agtttgggaa atgtccactc acccgtcagt ctataaatac ttagcccctc cctcattgtt
     1201 aagggagcaa aatctcagag agatagtcct agagagagaa agagagcaag tagcctagaa
     1261 gtagtcaagg cggcgaagta ttcaggcacg tggccaggaa gaagaaaagc caagacgacg
     1321 aaaacaggta agagctaagc ttcctgcagg ttcactgccg tataggcaga ttagaaccta
     1381 aaggaaccgg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
     1441 tgaaaaagtg gcaccgagtc ggtgcgttca ctgccgtata ggcagaattg ggaagataaa
     1501 gaccagtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
     1561 aaagtggcac cgagtcggtg cgttcactgc cgtataggca ggtttgagtc tcctaaatac
     1621 ggttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
     1681 tggcaccgag tcggtgcgtt cactgccgta taggcagatt atgtaactaa ggggccagtt
     1741 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
     1801 accgagtcgg tgcgttcact gccgtatagg cagtcacacg caaactggca gctgttttag
     1861 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
     1921 agtcggtgcg ttcactgccg tataggcagg ttcaaaggcc gctcgaggag ttttagagct
     1981 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
     2041 ggtgcgttca ctgccgtata ggcaggtcga tcgacaagct cgagtttctc cataataatg
     2101 tgtgagtagt tcccagataa gggaattagg gttcctatag ggtttcgctc atgtgttgag
     2161 catataagaa acccttagta tgtatttgta tttgtaaaat acttctatca ataaaatttc
     2221 taattcctaa aaccaaaatc cagtactaaa atccagatcc cccgaattag agctctaccg
     2281 gtgacgcagg tgatgaaaag ccttagggtc aggtggcact tttcggggaa atgtgcgatg
     2341 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctaa tgagacaata
     2401 accctgataa atgcttcaat tatattgaaa aaggaagagt atgagtatcc aacatttccg
     2461 tgtcgccctt attccctttt ttgcggcctt ctgccttcct gtttttgctc acccagaaac
     2521 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcc cgagtgggtt acattgaact
     2581 ggatctcaac agcggcaaaa tcttagagag ttttcgcccc gaagaacgct tcccaatgat
     2641 gagcactttc aaagttctgc tatgtggcgc ggtgttatcc cgtattgatg ctgggcaaga
     2701 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac
     2761 agaaaagcat cttacggatg gcatgacagt aagagaatta tgtagtgctg ccataaccat
     2821 gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac
     2881 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     2941 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3001 tctccgcaaa ctattaactg gcgaactact tactcttgct tcacgccagc aactcattga
     3061 ctggatggag gcggacaaag ttgcaggacc acttctgcgc tcggcacttc cggctggctg
     3121 gtttattgct gataaatctg gagccggtga gcgtggctct cgcggtatca ttgcagcact
     3181 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3241 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     3301 actgtcagac gaagggaaat aaatagtagc ccgcctgatg tgcgggcttt tttttggtgg
     3361 tggttaccag tgtcgcttag ctaccgtagg tcctcttagg cggtccgatt agcgacatag
     3421 tcatcaat
//
