LOCUS       sequence-393542-        3828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MODULE B tRNA vector with 6x gRNAs targeting the four MsPho2-1abcd 
            and MsPho2-2abcd genes in alfalfa (pMOD_B2303 - #91068).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3828)
  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 3828)
  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-tRNA
FEATURES             Location/Qualifiers
     source          1..3828
                     /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"
     misc_RNA        1448..1523
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        1621..1696
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        1794..1869
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        1967..2042
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        2140..2215
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        2313..2388
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     polyA_signal    2485..2659
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        2736..2840
                     /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 aacaaagcac cagtggtcta gtggtagaat
     1381 agtaccctgc cacggtacag acccgggttc gattcccggc tggtgcaatt agaacctaaa
     1441 ggaaccggtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
     1501 aaaaagtggc accgagtcgg tgcaacaaag caccagtggt ctagtggtag aatagtaccc
     1561 tgccacggta cagacccggg ttcgattccc ggctggtgca aattgggaag ataaagacca
     1621 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
     1681 ggcaccgagt cggtgcaaca aagcaccagt ggtctagtgg tagaatagta ccctgccacg
     1741 gtacagaccc gggttcgatt cccggctggt gcagtttgag tctcctaaat acggttttag
     1801 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
     1861 agtcggtgca acaaagcacc agtggtctag tggtagaata gtaccctgcc acggtacaga
     1921 cccgggttcg attcccggct ggtgcaatta tgtaactaag gggccagttt tagagctaga
     1981 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
     2041 gcaacaaagc accagtggtc tagtggtaga atagtaccct gccacggtac agacccgggt
     2101 tcgattcccg gctggtgcat cacacgcaaa ctggcagctg ttttagagct agaaatagca
     2161 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcaacaa
     2221 agcaccagtg gtctagtggt agaatagtac cctgccacgg tacagacccg ggttcgattc
     2281 ccggctggtg cagttcaaag gccgctcgag gagttttaga gctagaaata gcaagttaaa
     2341 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgcaa caaagcacca
     2401 gtggtctagt ggtagaatag taccctgcca cggtacagac ccgggttcga ttcccggctg
     2461 gtgcagtcga tcgacaagct cgagtttctc cataataatg tgtgagtagt tcccagataa
     2521 gggaattagg gttcctatag ggtttcgctc atgtgttgag catataagaa acccttagta
     2581 tgtatttgta tttgtaaaat acttctatca ataaaatttc taattcctaa aaccaaaatc
     2641 cagtactaaa atccagatcc cccgaattag agctctaccg gtgacgcagg tgatgaaaag
     2701 ccttagggtc aggtggcact tttcggggaa atgtgcgatg aacccctatt tgtttatttt
     2761 tctaaataca ttcaaatatg tatccgctaa tgagacaata accctgataa atgcttcaat
     2821 tatattgaaa aaggaagagt atgagtatcc aacatttccg tgtcgccctt attccctttt
     2881 ttgcggcctt ctgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2941 ctgaagatca gttgggtgcc cgagtgggtt acattgaact ggatctcaac agcggcaaaa
     3001 tcttagagag ttttcgcccc gaagaacgct tcccaatgat gagcactttc aaagttctgc
     3061 tatgtggcgc ggtgttatcc cgtattgatg ctgggcaaga gcaactcggt cgccgcatac
     3121 actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg
     3181 gcatgacagt aagagaatta tgtagtgctg ccataaccat gagtgataac actgcggcca
     3241 acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3301 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3361 acgagcgtga caccacgatg cctgtagcaa tggcaacaac tctccgcaaa ctattaactg
     3421 gcgaactact tactcttgct tcacgccagc aactcattga ctggatggag gcggacaaag
     3481 ttgcaggacc acttctgcgc tcggcacttc cggctggctg gtttattgct gataaatctg
     3541 gagccggtga gcgtggctct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3601 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3661 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac gaagggaaat
     3721 aaatagtagc ccgcctgatg tgcgggcttt tttttggtgg tggttaccag tgtcgcttag
     3781 ctaccgtagg tcctcttagg cggtccgatt agcgacatag tcatcaat
//
