LOCUS       sequence-252670-        5649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level0 StCas9, Arabidopsis codon optimized.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5649)
  AUTHORS   Hahn F, Korolev A, Sanjurjo Loures L, Nekrasov V
  TITLE     A modular cloning toolkit for genome editing in plants.
  JOURNAL   BMC Plant Biol. 2020 Apr 23;20(1):179. doi: 
            10.1186/s12870-020-02388-2.
  PUBMED    32326901
REFERENCE   2  (bases 1 to 5649)
  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: "BMC Plant 
            Biol."; date: "2020-04-23"; volume: "20(1):179. doi"; pages: " 
            10.1186/s12870-020-02388-2"
COMMENT     Sequence Label: pFH15
FEATURES             Location/Qualifiers
     source          1..5649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..42)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        110..214
                     /gene="bla"
                     /label=AmpR promoter
     CDS             436..1227
                     /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      1398..1986
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1887..1906
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2140..2157
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             5610..5630
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gtggctcttc agtggacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
       61 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
      121 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
      181 ataaatgctt caataatatt gaaaaaggaa gagtatgcgc tcacgcaact ggtccagaac
      241 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg
      301 tttttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg
      361 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa
      421 acaaagttaa acatcatgag ggaagcggtg atcgccgaag tatcgactca actatcagag
      481 gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc
      541 tccgcagtgg atggcggcct gaagccacac agcgatattg atttgctggt tacggtgacc
      601 gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct
      661 tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac
      721 atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat
      781 gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg
      841 acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat
      901 ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg
      961 ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac
     1021 agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc
     1081 ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa
     1141 gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccatta cgtaaaaggc
     1201 gagatcacca aggtagtcgg caaataactg tcagaccaag tttactcata tatactttag
     1261 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1321 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1381 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1441 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1501 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1561 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1621 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1681 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1741 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1801 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1861 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1921 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     1981 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2041 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2101 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2161 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatca
     2221 ctctgtggtc tcaaatgtct gatctcgtgc tcggactcga tatcggaatc ggatctgttg
     2281 gagtgggaat cctcaacaag gttaccggtg agatcatcca caagaactct aggatcttcc
     2341 ctgctgctca ggctgagaac aacctcgtga gaagaactaa caggcagggt agaaggctcg
     2401 ctagaagaaa gaagcacaga agggttaggc tcaacagact cttcgaggaa tctggactca
     2461 tcaccgattt caccaagatc tctatcaacc tcaaccctta ccagctcagg gtgaagggac
     2521 ttaccgatga gctttctaac gaggaactct tcattgctct caagaacatg gtgaagcaca
     2581 ggggtatcag ttacctcgat gatgcttctg atgatggaaa ctcttctgtg ggagattacg
     2641 ctcagatcgt gaaagagaac tctaagcagc tcgagactaa gacccctgga caaatccagc
     2701 ttgagagata ccagacttac ggacagctta ggggagattt caccgttgag aaggatggaa
     2761 agaaacacag gctcatcaac gtgttcccta cctctgctta cagatctgag gctctcagaa
     2821 tcctccagac ccagcaagaa ttcaaccctc agatcaccga tgagttcatc aacagatacc
     2881 tcgagatcct cactggtaag aggaagtact atcacggacc tggaaacgag aagtctagga
     2941 ccgattacgg aaggtacaga acctctggtg agactctcga taacatcttc ggtatcctca
     3001 tcggaaagtg cacattctac ccagatgaat tcagggctgc taaggcttct tacaccgctc
     3061 aagagttcaa cctcctcaac gatctcaaca acctcactgt gcctaccgag acaaagaagc
     3121 tctctaaaga gcagaagaac cagatcatca actacgtgaa gaacgagaag gctatgggac
     3181 ctgctaagct tttcaagtat atcgctaagc tcctctcttg tgatgtggct gatatcaagg
     3241 gatacaggat cgataagtct ggaaaggctg agatccacac cttcgaggct tacagaaaga
     3301 tgaaaaccct cgaaaccttg gatatcgagc agatggatag ggaaaccctc gataagctcg
     3361 cttacgtgct cactctcaac actgagagag agggaatcca agaggctctc gaacatgagt
     3421 tcgctgatgg atctttctca caaaagcaag tggatgagct tgtgcagttc agaaaggcta
     3481 actcttctat cttcggaaag ggatggcaca acttctctgt gaagcttatg atggaactta
     3541 tccctgagct ttacgagact tctgaggaac agatgaccat cctcaccagg ctcggaaagc
     3601 aaaagaccac ctcttcatct aacaagacca agtatattga tgagaagctc ctcaccgaag
     3661 agatctacaa ccctgtggtt gctaagtctg tgaggcaggc tatcaagatc gttaacgctg
     3721 ctatcaaaga atacggtgat tttgataaca tcgtgatcga gatggcaagg gaaacaaacg
     3781 aggatgatga gaaaaaggct atccaaaaga tccagaaagc taacaaggat gagaaggatg
     3841 ctgctatgct caaggctgct aaccagtaca atggtaaggc tgagcttcct cactctgtgt
     3901 tccatggtca taagcagctt gctaccaaga tcagactctg gcatcaacag ggtgagagat
     3961 gcctctacac tggaaagacc atctctatcc atgatctcat caacaactct aaccagttcg
     4021 aagtggatca catcctccca ctctcaatca ccttcgatga ttctctcgca aacaaggtgc
     4081 tcgtgtacgc taccgctaat caagagaagg gacagaggac tccataccag gcactcgatt
     4141 ctatggatga tgcatggtca ttcagggaac tcaaggcttt cgtgagagag tctaagaccc
     4201 tcagtaacaa gaaaaaagag tacctcctca ctgaggaaga tatctctaag ttcgatgtga
     4261 ggaagaagtt cattgagagg aacctcgtgg ataccaggta cgcttctaga gttgtgctca
     4321 acgctctcca agagcatttc agggctcaca agatcgatac aaaggtgtca gtggtgaggg
     4381 gacagttcac ttctcagctt agaaggcact ggggaatcga aaagaccagg gatacttacc
     4441 atcaccacgc tgtggatgct cttattatcg ctgcttcatc tcagctcaac ctctggaaga
     4501 agcagaaaaa caccctcgtg tcttactctg aggatcagct cctcgatatt gagactggtg
     4561 agcttatctc agatgatgag tacaaagagt cagttttcaa ggcaccatac cagcatttcg
     4621 ttgataccct caagtctaaa gagttcgagg attctatcct cttcagttac caagtggatt
     4681 ctaagttcaa tagaaagatc tctgatgcta ccatctacgc taccaggcaa gctaaggtgg
     4741 gaaaggataa ggctgatgag acatacgtgc tcggtaagat caaggatatc tacacccagg
     4801 atggatacga tgctttcatg aagatctata agaaggataa gtctaagttc ctcatgtaca
     4861 ggcacgatcc acagacattc gagaaggtga tcgagcctat cctcgagaac taccctaaca
     4921 agcagatcaa cgataaggga aaagaagtgc cttgcaaccc attcctcaag tacaaagaag
     4981 aacacggtta catcagaaag tactctaaga agggaaacgg acctgagatc aagtctctca
     5041 agtactacga ttctaagctc ggaaaccaca tcgatatcac ccctaagaat tctaacaaca
     5101 aggtggtgct ccagtctgtg tctccttgga gggctgatgt gtacttcaac aagactaccg
     5161 gaaagtacga gatccttgga ctcaagtacg ctgatctcca gttcgataag ggtactggaa
     5221 cctacaagat ctcacaagaa aagtacaacg atatcaaaaa gaaagagggt gttgattctg
     5281 attcagagtt caagttcacc ctctacaaga acgatttgct cctcgtgaag gatactgaga
     5341 caaaagagca gcagcttttc agattccttt ctaggaccat gcctaagcag aagcactacg
     5401 ttgagcttaa gccttacgat aagcaaaagt tcgagggtgg tgaggcactc atcaaggttc
     5461 tcggaaacgt tgcaaactct ggacagtgca agaagggact cggaaagtct aacatctcaa
     5521 tctacaaggt gaggaccgat gtgcttggaa accagcacat catcaagaat gagggtgata
     5581 agcctaagct cgatttctca agggctgatc ctaagaagaa gaggaaggtt tgagctttga
     5641 gaccacgaa
//
