LOCUS       sequence-308835-        8407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for PCR amplification of the constant part of guide RNAs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8407)
  AUTHORS   Grutzner R, Martin P, Horn C, Mortensen S, Cram EJ, Lee-Parsons CWT,
            Stuttmann J, Marillonnet S
  TITLE     High-efficiency genome editing in plants mediated by a Cas9 gene 
            containing multiple introns.
  JOURNAL   Plant Commun. 2020 Nov 23;2(2):100135. doi: 
            10.1016/j.xplc.2020.100135. eCollection 2021 Mar 8.
  PUBMED    33898975
REFERENCE   2  (bases 1 to 8407)
  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: "Plant 
            Commun."; date: "2020-11-23"; volume: "2(2):100135. doi"; pages: " 
            10.1016/j.xplc.2020.100135. eCollection 2021 Mar 8"
COMMENT     Sequence Label: pAGM9037
FEATURES             Location/Qualifiers
     source          1..8407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        6076..6151
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(6198..6216)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6284..6388
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6610..7401
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      7572..8160
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8061..8080
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8314..8331
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 attgtgagac gtgccggtac cgcacggtct gccaatccga cggaggttta tgaattttcc
       61 accttttcca caagctcaac tagtattaac gatgtggatt tagcaaaaaa aacctgtaac
      121 cctaaatgta aaataacggg taagcctgcc aaccatgtta tggcagatta agcgtctttt
      181 tgaagggcac cgcatctttc gcgttgccgt aaatgtatcc gtttagcaat gaggattagt
      241 gtaatgaata atccgtcgtt actcaatcat gcggtcgaaa cgatggcagt tggctcgaaa
      301 agttttgcga cagcctcaaa gttatttgat gcaaaaaccc ggcgcagcgt actgatgctc
      361 tacgcctggt gccgccattg tgacgatgtt attgacgatc agacgctggg ctttcaggcc
      421 cggcagcctg ccttacaaac gcccgaacaa cgtctgatgc aacttgagat gaaaacgcgc
      481 caggcctatg caggatcgca gatgcacgaa ccggcgtttg cggcttttca ggaagtggct
      541 atggctcatg atatcgcccc ggcttacgcg tttgatcatc tggaaggctt cgccatggat
      601 gtacgcgaag cgcaatacag ccaactggat gatacgctgc gctattgcta tcacgttgca
      661 ggcgttgtcg gcttgatgat ggcgcaaatc atgggcgtgc gggataacgc cacgctggac
      721 cgcgcctgtg accttgggct ggcatttcag ttgaccaata ttgctcgcga tattgtggac
      781 gatgcgcatg cgggccgctg ttatctgccg gcaagctggc tggagcatga aggtctgaac
      841 aaagagaatt atgcggcacc tgaaaaccgt caggcgctga gccgtatcgc ccgtcgtttg
      901 gtgcaggaag cagaacctta ctatttgtct gccacagccg gcctggcagg gttgcccctg
      961 cgttccgcct gggcaatcgc tacggcgaag caggtttacc ggaaaatagg tgtcaaagtt
     1021 gaacaggccg gtcagcaagc ctgggatcag cggcagtcaa cgaccacgcc cgaaaaatta
     1081 acgctgctgc tggccgcctc tggtcaggcc cttacttccc ggatgcgggc tcatcctccc
     1141 cgccctgcgc atctctggca gcgcccgctc taggcttact agaaggaggt tatcaatgaa
     1201 accaactacg gtaattggtg caggcttcgg tggcctggca ctggcaattc gtctacaagc
     1261 tgcggggatc cccgtcttac tgcttgaaca acgtgataaa cccggcggtc gggcttatgt
     1321 ctacgaggat caggggttta cctttgatgc aggcccgacg gttatcaccg atcccagtgc
     1381 cattgaagaa ctgtttgcac tggcaggaaa acagttaaaa gagtatgtcg aactgctgcc
     1441 ggttacgccg ttttaccgcc tgtgttggga gtcagggaag gtctttaatt acgataacga
     1501 tcaaacccgg ctcgaagcgc agattcagca gtttaatccc cgcgatgtcg aaggttatcg
     1561 tcagtttctg gactattcac gcgcggtgtt taaagaaggc tatctaaagc tcggtactgt
     1621 ccctttttta tcgttcagag acatgcttcg cgccgcacct caactggcga aactgcaggc
     1681 atggagaagc gtttacagta aggttgccag ttacatcgaa gatgaacatc tgcgccaggc
     1741 gttttctttc cactcgctgt tggtgggcgg caatcccttc gccacctcat ccatttatac
     1801 gttgatacac gcgctggagc gtgagtgggg cgtctggttt ccgcgtggcg gcaccggcgc
     1861 attagttcag gggatgataa agctgtttca ggatctgggt ggcgaagtcg tgttaaacgc
     1921 cagagtcagc catatggaaa cgacaggaaa caagattgaa gccgtgcatt tagaggacgg
     1981 tcgcaggttc ctgacgcaag ccgtcgcgtc aaatgcagat gtggttcata cctatcgcga
     2041 cctgttaagc cagcaccctg ccgcggttaa gcagtccaac aaactgcaga ctaagcgcat
     2101 gagtaactct ctgtttgtgc tctattttgg tttgaatcac catcatgatc agctcgcgca
     2161 tcacacggtt tgtttcggcc cgcgttaccg cgagctgatt gacgaaattt ttaatcatga
     2221 tggcctcgca gaggacttct cactttatct gcacgcgccc tgtgtcacgg attcgtcact
     2281 ggcgcctgaa ggttgcggca gttactatgt gttggcgccg gtgccgcatt taggcaccgc
     2341 gaacctcgac tggacggttg aggggccaaa actacgcgac cgtatttttg cgtaccttga
     2401 gcagcattac atgcctggct tacggagtca gctggtcacg caccggatgt ttacgccgtt
     2461 tgattttcgc gaccagctta atgcctatca tggctcagcc ttttctgtgg agcccgttct
     2521 tacccagagc gcctggtttc ggccgcataa ccgcgataaa accattacta atctctacct
     2581 ggtcggcgca ggcacgcatc ccggcgcagg cattcctggc gtcatcggct cggcaaaagc
     2641 gacagcaggt ttgatgctgg aggatctgat atgagctttt actaaggagc cctgaaatga
     2701 cggtctgcgc aaaaaaacac gttcatctca ctcgcgatgc tgcggagcag ttactggctg
     2761 atattgatcg acgccttgat cagttattgc ccgtggaggg agaacgggat gttgtgggtg
     2821 ccgcgatgcg tgaaggtgcg ctggcaccgg gaaaacgtat tcgccccatg ttgctgttgc
     2881 tgaccgcccg cgatctgggt tgcgctgtca gccatgacgg attactggat ttggcctgtg
     2941 cggtggaaat ggtccacgcg gcttcgctga tccttgacga tatgccctgc atggacgatg
     3001 cgaagctgcg gcgcggacgc cctaccattc attctcatta cggagagcat gtggcaatac
     3061 tggcggcggt tgccttgctg agtaaagcct ttggcgtaat tgccgatgca gatggcctca
     3121 cgccgctggc aaaaaatcgg gcggtttctg aactgtcaaa cgccatcggc atgcaaggat
     3181 tggttcaggg tcagttcaag gatctgtctg aaggggataa gccgcgcagc gctgaagcta
     3241 ttttgatgac gaatcacttt aaaaccagca cgctgttttg tgcctccatg cagatggcct
     3301 cgattgttgc gaatgcctcc agcgaagcgc gtgattgcct gcatcgtttt tcacttgatc
     3361 ttggtcaggc atttcaactg ctggacgatt tgaccgatgg catgaccgac accggtaagg
     3421 atagcaatca ggacgccggt aaatcgacgc tggtcaatct gttaggcccg agggcggttg
     3481 aagaacgtct gagacaacat cttcagcttg ccagtgagca tctctctgcg gcctgccaac
     3541 acgggcacgc cactcaacat tttattcagg cctggtttga caaaaaactc gctgccgtca
     3601 gttaagcttc agagaaggat gtctacaatg agttttgata ttgccaaata cccgaccctg
     3661 gcactggtcg actccaccca ggagttacga ctgttgccga aagagagttt accgaaactc
     3721 tgcgacgaac tgcgccgcta tttactcgac agcgtgagcc gttccagcgg gcacttcgcc
     3781 tccgggctgg gcacggtcga actgaccgtg gcgctgcact atgtctacaa caccccgttt
     3841 gaccaattga tttgggatgt ggggcatcag gcttatccgc ataaaatttt gaccggacgc
     3901 cgcgacaaaa tcggcaccat ccgtcagaaa ggcggtctgc acccgttccc gtggcgcggc
     3961 gaaagcgaat atgacgtatt aagcgtcggg cattcatcaa cctccattag tgccggaatt
     4021 ggtattgcgg ttgctgccga aaaagaaggc aaaaatcgcc gcaccgtctg tgtcattggc
     4081 gatggcgcga ttaccgcagg catggcgttt gaagcgatga atcacgcggg cgatatccgt
     4141 cctgatatgc tggtgattct caacgacaat gaaatgtcga tttccgaaaa tgtcggcgcg
     4201 ctcaacaacc atctggcaca gctgctttcc ggtaagcttt actcttcact gcgcgaaggc
     4261 gggaaaaaag ttttctctgg cgtgccgcca attaaagagc tgctcaaacg caccgaagaa
     4321 catattaaag gcatggtagt gcctggcacg ttgtttgaag agctgggctt taactacatc
     4381 ggcccggtgg acggtcacga tgtgctgggg cttatcacca cgctaaagaa catgcgcgac
     4441 ctgaaaggcc cgcagttcct gcatatcatg accaaaaaag gtcgtggtta tgaaccggca
     4501 gaaaaagacc cgatcacttt ccacgccgtg cctaaatttg atccctccag cggttgtttg
     4561 ccgaaaagta gcggcggttt gccgagctat tcaaaaatct ttggcgactg gttgtgcgaa
     4621 acggcagcga aagacaacaa gctgatggcg attactccgg cgatgcgtga aggttccggc
     4681 atggtcgagt tttcacgtaa attcccggat cgctacttcg acgtggcaat tgccgagcaa
     4741 cacgcggtga cctttgctgc gggtctggcg attggtgggt acaaacccat tgtcgcgatt
     4801 tactccactt tcctgcaacg cgcctatgat caggtgctgc atgacgtggc gattcaaaag
     4861 cttccggtcc tgttcgccat cgaccgcgcg ggcattgttg gtgctgacgg tcaaacccat
     4921 cagggtgctt ttgatctctc ttacctgcgc tgcataccgg aaatggtcat tatgaccccg
     4981 agcgatgaaa acgaatgtcg ccagatgctc tataccggct atcactataa cgatggcccg
     5041 tcagcggtgc gctacccgcg tggcaacgcg gtcggcgtgg aactgacgcc gctggaaaaa
     5101 ctaccaattg gcaaaggcat tgtgaagcgt cgtggcgaga aactggcgat ccttaacttt
     5161 ggtacgctga tgccagaagc ggcgaaagtc gccgaatcgc tgaacgccac gctggtcgat
     5221 atgcgttttg tgaaaccgct tgatgaagcg ttaattctgg aaatggccgc cagccatgaa
     5281 gcgctggtca ccgtagaaga aaacgccatt atgggcggcg caggcagcgg cgtgaacgaa
     5341 gtgctgatgg cccatcgtaa accagtaccc gtgctgaaca ttggcctgcc ggacttcttt
     5401 attccgcaag gaactcagga agaaatgcgc gccgaactcg gcctcgatgc cgctggtatg
     5461 gaagccaaaa tcaaggcctg gctggcataa gctttgtgtg aggagaatga aaatgcaaac
     5521 ggaacacgtc attttattga atgcacaggg agttcccacg ggtacgctgg aaaagtatgc
     5581 cgcacacacg gcagacaccc gcttacatct cgcgttctcc agttggctgt ttaatgccaa
     5641 aggacaatta ttagttaccc gccgcgcact gagcaaaaaa gcatggcctg gcgtgtggac
     5701 taactcggtt tgtgggcacc cacaactggg agaaagcaac gaggacgcag tgatccgccg
     5761 ttgccgttat gagcttggcg tggaaattac gcctcctgaa tctatctatc ctgactttcg
     5821 ctaccgcgcc accgatccga gtggcattgt ggaaaatgaa gtgtgtccgg tatttgccgc
     5881 acgcaccact agtgcgttac agatcaatga tgatgaagtg atggattatc aatggtgtga
     5941 tttagcagat gtattacacg gtattgatgc cacgccgtgg gcgttcagtc cgtggatggt
     6001 gatgcaggcg acaaatcgcg aagccagaaa acgattatct gcatttaccc agcttaaata
     6061 agcttgagcg tctcagtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
     6121 tcaacttgaa aaagtggcac cgagtcggtg ctttttttcg cttgagacca cgaagtggct
     6181 cttcagtgga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     6241 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     6301 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     6361 gcttcaataa tattgaaaaa ggaagagtat gcgctcacgc aactggtcca gaaccttgac
     6421 cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt
     6481 tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga
     6541 tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag
     6601 ttaaacatca tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt
     6661 ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca
     6721 gtggatggcg gcctgaagcc acacagcgat attgatttgc tggttacggt gaccgtaagg
     6781 cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct
     6841 ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt
     6901 ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt
     6961 cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa
     7021 gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt
     7081 cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc
     7141 gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca
     7201 gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg
     7261 gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca agaagaagat
     7321 cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc attacgtaaa aggcgagatc
     7381 accaaggtag tcggcaaata actgtcagac caagtttact catatatact ttagattgat
     7441 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     7501 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     7561 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     7621 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     7681 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     7741 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     7801 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     7861 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7921 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7981 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     8041 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     8101 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     8161 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     8221 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     8281 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     8341 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atcactctgt
     8401 ggtctca
//
