LOCUS       sequence-307875-        7862 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 zCas9i vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7862)
  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 7862)
  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: pAGM47523
FEATURES             Location/Qualifiers
     source          1..7862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             301..1092
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1433..2021
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1922..1941
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2086..2106
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7839..7859
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gctttgagac cacgaagtgc cgaggcacga acccagtgga cataagcctg ttcggttcgt
       61 aagctgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt gaccgaacgc
      121 agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttggggta
      181 cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg
      241 ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatc
      301 atgggggaag cggtgatcgc cgaagtatcg actcaactat cagaggtagt tggcgtcatc
      361 gagcgccatc tcgaaccgac gttgctggcc gtacatttgt acggctccgc agtggatggc
      421 ggcctgaagc cacacagcga tattgatttg ctggttacgg tgaccgtaag gcttgatgaa
      481 acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc tggagagagc
      541 gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat tccgtggcgt
      601 tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat tcttgcaggt
      661 atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa agcaagagaa
      721 catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt tcctgaacag
      781 gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc cgactgggct
      841 ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc agtaaccggc
      901 aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat
      961 cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga tcgcttggcc
     1021 tcgcgcgcag atcagttgga agaatttgtc cattacgtga aaggcgagat caccaaggta
     1081 gtcggcaaat aatgtctagc tagaaattcg ttcaagccga cgccgcttcg cggcgcggct
     1141 taactcaagc gttagatgca ctaagcacat aattgctcac agccaaacta tcaggtcaag
     1201 tctgctttta ttatttttaa gcgtgcataa taagccctac acaaattggg agatatatca
     1261 tgctgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     1321 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     1381 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1441 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1501 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     1561 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     1621 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1681 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1741 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     1801 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     1861 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     1921 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     1981 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2041 ttttacggtt cctgcactct gtggtctcaa atggcttcta gcccaccgaa gaagaagcgg
     2101 aaggtcagct ggaaaatgga caagaagtac agcattggac ttgatattgg tacgaactca
     2161 gttgggtggg ccgttatcac cgatgaatac aaggtacctt cgaagaaatt taaagtgctg
     2221 ggcaacacag ataggcacag cattaagaag aacttgatcg gagctctgct ctttgactct
     2281 ggagaaaccg cggaggcgac aaggcttaaa cgtactgcga ggagaaggta cactcgcagg
     2341 aagaacagaa tctgttatct ccaagagatc tttagcaacg agatggcgaa ggtaaggatt
     2401 tttatgatat agtatgctta tgtattttgt actgaaagca tatcctgctt cattgggata
     2461 ttactgaaag catttaacta catgtaaact cacttgatga tcaataaact tgattttgca
     2521 ggttgacgac tcgttcttcc atcgcctcga ggaatccttc ctggtagagg aagataagaa
     2581 acacgagcgt caccccatct ttgggaatat tgttgacgaa gtagcctatc atgaaaagta
     2641 tccgactata taccaccttc gcaagaagct ggtggactca accgataagg cagaccttcg
     2701 gctcatatac ctggctctcg cgcacatgat aaagtttcgt ggccatttct tgatcgaagg
     2761 ggacctcaac ccggataact ccgatgtgga taaactgttc attcagctcg tccaaaccta
     2821 caatcagctg ttcgaggaga accccatcaa tgcatcaggt aacattcctt agttaccttt
     2881 cttttctttt tccatcataa gtttatagat tgtacatgct ttgagatttt tctttgcaaa
     2941 caatctcagg tgtcgacgcc aaggcaatac tgtctgccag actttcgaag tccagacggc
     3001 ttgagaatct gatcgctcaa ttgccaggcg agaagaagaa cggcttgttc gggaatctga
     3061 ttgcactgtc tctgggcctc acccctaact tcaaaagcaa ctttgacctc gccgaggacg
     3121 cgaagctgca gctgtcaaag gatacatacg atgatgatct ggacaatctg ctcgcccaaa
     3181 taggtgctct tgaaattgga actcttcttt tgttgtctaa acctatcaat ttctttgcgg
     3241 aaatttattt gaagctgtag agttaaaatt gagtctttta aacttttgta ggtgatcagt
     3301 atgccgacct gttcttggct gccaagaatc tgtcagacgc tatcttgctc agtgacattc
     3361 tgcgggtcaa cacggagata accaaagcgc cacttagcgc ctccatgatc aagaggtacg
     3421 acgagcatca ccaggatctg acccttctga aggctttggt tcgccagcaa ctccccgaga
     3481 agtacaagga gattttcttt gaccaatcga agaatggcta cgcagggtac attgatggag
     3541 gtaagttgtt acttatgatt gttttcctct ctgctacatg tattttgttg ttcatttctg
     3601 taagatataa gaattgagtt ttcctctgat gatattatta ggtgcaagtc aggaggaatt
     3661 ctacaaattc atcaagccta ttctggaaaa gatggacggt acagaggagc tgctcgttaa
     3721 attgaaccgc gaagatttgc ttcggaagca gcgtaccttc gacaatggca gcataccgca
     3781 ccagatccac ctcggtgagc tgcatgctat cttgaggagg caagaggact tctatccgtt
     3841 cctgaaagac aacagagaga agattgaaaa gatcctcacg ttccgcattc cctactatgt
     3901 aggttagtat catatgaaga aatacctagt ttcagttgat gaatgctatt ttctgacctc
     3961 agttgttctc ttttgagaat tatttctttt ctaatttgcc tgatttttct attaattcat
     4021 taggtccact cgcacgcggg aactcgcggt ttgcgtggat gacacgcaaa tccgaggaga
     4081 ctatcacgcc ttggaacttc gaagaggtcg tggacaaggg tgcgagtgca cagtccttca
     4141 tcgaaaggat gaccaacttc gataagaatc tcccaaatga gaaagtcctg cccaagcata
     4201 gtctcctgta cgaatacttc acggtctaca acgagctgac gaaggtgaaa tatgtgacgg
     4261 aggggatgcg caaaccggcc ttcctgtcag gtaaatcctg gtccacactt ttacgataaa
     4321 aacacaagat tttaaactat gaactgatca ataatcattc ctaaaagacc acacttttgt
     4381 tttgtttcta aagtaatttt tactgttata acaggtgagc agaagaaggc cattgtcgat
     4441 ctcttgttca aaaccaatcg gaaggtcact gtgaaacagc ttaaagagga ctactttaag
     4501 aagatcgaat gctttgattc tgtggaaatc agcggcgttg aggataggtt caatgcctct
     4561 cttggcacat accatgacct gttgaaaatc atcaaggaca aggacttcct tgacaacgag
     4621 gagaacgagg acatcctcga ggacatcgtg ctgactctca cgctgtttga ggacagagaa
     4681 atgatcgagg agcgccttaa gacttatgcg catctgttcg atgacaaggt catgaagcag
     4741 ttgaagagga ggagatatac aggtaagagg tcaaaaggtt tccgcaatga tccctctttt
     4801 tttgtttctc tagtttcaag aatttgggta tatgactaac ttctgagtgt tccttgatgc
     4861 atatttgtga tgagacaaat gtttgttcta tgttttaggt tggggaaggc tctccaggaa
     4921 gctcatcaac ggcatccgcg acaagcaatc cggcaagact atactggact ttctcaaatc
     4981 cgacggtttt gcgaatcgga acttcatgca gcttattcac gatgactcac tgaccttcaa
     5041 agaagatatc cagaaggccc aagtgtcagg tcagggcgat agccttcacg aacacatagc
     5101 caacctggct ggatcgccag ctataaagaa gggcatactg cagacagtga aggttgtgga
     5161 tgagctggtg aaggtaagtt ctgcatttgg ttatgctcct tgcattttag gtgttcgtcg
     5221 cacttccatt tccatgaata gctaagattt tttttctctg cattcattct tcttgcctca
     5281 gttctaactg tttgtggtat ttttgtttta attattgcta caggtcatgg gccgccataa
     5341 gccggagaac atcgtcatcg agatggcgag ggaaaaccag acgactcaga aagggcagaa
     5401 gaactcacgg gagcgcatga agcggataga ggaaggcatc aaggagcttg ggagtcagat
     5461 tctgaaagag cacccagtcg aaaatactca actccagaac gagaagctgt acctctatta
     5521 cctccagaat gggagagata tgtacgtcga ccaagagctc gacattaaca gactctccga
     5581 ctatgatgtg gatcacattg tccctcaatc tttcctgaag gacgatagta ttgacaacaa
     5641 ggtaaagcaa ctgtgtttta atcaatttct tgtcaggata tatggattat aacttaattt
     5701 ttgagaaatc tgtagtattt ggcgtgaaat gagtttgctt tttggtttct cccgtgttat
     5761 aggtccttac gcgctcagac aagaaccgcg gaaaatccga caatgtaccc agcgaggagg
     5821 ttgtgaagaa gatgaagaac tattggaggc agcttttgaa tgctaagctc ataacccaac
     5881 ggaaattcga caatctcacg aaggcagaaa ggggcggact gtctgagctc gacaaagccg
     5941 gcttcatcaa gcgccagttg gttgaaactc gtcagattac gaaacatgtg gcccagatac
     6001 tcgattcgcg tatgaatacg aagtatgatg agaatgacaa acttatcagg gaggtaaagg
     6061 taaagtttcc aactttcctt taccatatca aactaaagtt cgaaactttt tatttgatca
     6121 acttcaaggc cacccgatct ttctattcct gattaatttg tgatgaatcc atattgactt
     6181 ttgatggtta cgcaggtgat caccctcaag agcaaactgg ttagtgactt ccggaaggac
     6241 ttccagtttt acaaggttcg cgagatcaac aactaccatc atgcccatga cgcctacctg
     6301 aacgccgttg ttggcactgc tctcatcaag aagtatccga aactggagtc tgagtttgtg
     6361 tacggggatt acaaggtgta cgacgttagg aagatgatcg cgaagtcaga acaagagatc
     6421 ggcaaggcta ccgcgaaata cttcttttac tcgaatatca tgaacttctt caagacagag
     6481 atcactctgg cgaatggtga aatccggaag aggcctctga tcgagacaaa tggcgaaaca
     6541 ggtctgtctt tcctatttca tatgtttaat cctaggaatt tgatcaattg attgtatgta
     6601 tgtcgatccc aagactttct tgttcactta tatcttaact ctctctttgc tgtttcttgc
     6661 aggtgagatt gtctgggata agggcaggga ttttgcgact gtgcgtaagg ttctcagcat
     6721 gccccaagtc aacatagtca agaaaacgga ggttcaaacc ggtggtttct ccaaggagtc
     6781 cattctccct aagcgcaact ccgacaaact gattgcgagg aagaaggatt gggatccgaa
     6841 gaaatacgga ggctttgata gccctaccgt ggcatacagc gtactggtag tggccaaggt
     6901 ggagaagggc aagagcaaga aactgaaaag cgtcaaggaa ctgcttggaa ttaccataat
     6961 ggaaaggtcc tcgttcgaga agaatccgat cgacttcctc gaggctaaag gtaaaatatt
     7021 ggatgccaga cgatattctt tcttttgatt tgtaactttt tcctgtcaag gtcgataaat
     7081 tttatttttt ttggtaaaag gtcgataatt tttttttgga gccattatgt aattttccta
     7141 attaactgaa ccaaaattat acaaaccagg ttacaaagag gtgaagaaag acctcattat
     7201 caaactgccc aagtattcgc ttttcgaatt ggaaaatggc agaaaacgca tgctggcatc
     7261 tgccggagaa ctgcagaagg gcaacgagct ggcattgccc agtaagtacg tcaacttcct
     7321 gtacttggcc tcacactatg agaagctgaa ggggtcacca gaggacaacg agcagaagca
     7381 gttgtttgtc gagcagcaca agcactatct tgatgagatc atagagcaga tcagcgaatt
     7441 ttccaagcgg gtcattcttg cagacgctaa cctcgataag gtaaggactt ctcatgaata
     7501 ttagtggcag attagtgttg ttaaagtctt tggttagata atcgatgcct cctaattgtc
     7561 catgttttac tggttttcta caattaaagg tgctttccgc gtacaacaag cacagagata
     7621 agccgataag ggaacaagcg gaaaacatca tccacctgtt cacactgacc aatctgggag
     7681 ccccagcagc ctttaagtac ttcgatacca ctatcgacag aaagcgctac acatcaacca
     7741 aggaagtgtt ggacgctacc cttattcacc aatctattac agggctctat gagacaagga
     7801 tagatctgtc gcagttgggt ggtgactcta gggctgaccc aaagaagaag cgtaaagtct
     7861 ga
//
