LOCUS       sequence-409180-        7053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  L0 MoClo-compatible GoldenGate-module of dttLbCas12a-i for in planta
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7053)
  AUTHORS   Schindele P, Merker L, Schreiber T, Prange A, Tissier A, Puchta H
  TITLE     Enhancing gene editing and gene targeting efficiencies in 
            Arabidopsis thaliana by using an intron-containing version of 
            ttLbCas12a.
  JOURNAL   Plant Biotechnol J. 2023 Mar;21(3):457-459. doi: 10.1111/pbi.13964. 
            Epub 2022 Nov 30.
  PUBMED    36382936
REFERENCE   2  (bases 1 to 7053)
  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"; doi: "10.1111/pbi.13964";
            journalName: "Plant Biotechnol J"; date: "2023-03"; volume: "21"; 
            issue: "3"; pages: "457-459"
COMMENT     Sequence Label: pAGT8162 (AATG_dttLbCas12a-i_TTCG)
FEATURES             Location/Qualifiers
     source          1..7053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2089..2136
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             2439..3230
                     /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      3571..4159
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4060..4079
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             4218..4238
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gcaatgatcc ctcttttttt gtttctctag tttcaagaat ttgggtatat gactaacttc
       61 tgagtgttcc ttgatgcata tttgtgatga gacaaatgtt tgttctatgt tttaggtaag
      121 ctgtacatgt tccaaatcta taacaaggac ttctccgaca agtctcacgg cactcctaat
      181 ctgcatacaa tgtacttcaa gctgctgttc gacgagaaca accacggtca gattaggctt
      241 tctggtggtg ctgagctgtt catgagaagg gcctcactga agaaagaaga gttggtcgtt
      301 caccctgcca actctccaat cgctaacaag aaccctgaca acccgaaaaa gaccaccacc
      361 ttgtcttacg acgtgtacaa ggataagcgg ttcagcgagg atcagtacga gcttcacatt
      421 ccgatcgcca tcaacaagtg cccgaagaac atcttcaaga tcaataccga ggtgcgggtg
      481 ctgctgaagc acgatgataa tccttacgtg atcggcatcg ataggggcga gagaaacctt
      541 ctttacatcg tggtggtgga cggcaagggc aatatcgttg agcagtactc tctgaacgag
      601 attatcaaca atttcaacgg catccggatc aagaccgact accactctct gctggataag
      661 aaagaaaaag agcggttcga ggccaggcag aactggactt ctatcgaaaa catcaaagag
      721 ctgaaggccg gctacatctc tcaggtggtg cataagattt gcgagctggt ggaaaagtac
      781 gacgctgtga ttgctctcca ggatctgaac agcggcttca agaactcacg tgtgaaggta
      841 aagcaactgt gttttaatca atttcttgtc aggatatatg gattataact taatttttga
      901 gaaatctgta gtatttggcg tgaaatgagt ttgctttttg gtttctcccg tgttataggt
      961 tgagaagcag gtctaccaaa agttcgagaa gatgctcatc gacaagctga actacatggt
     1021 ggacaaaaag agcaaccctt gcgctaccgg tggtgctctt aagggttacc agatcactaa
     1081 caagttcgag tctttcaaga gcatgagcac ccagaacggc ttcatcttct acatccctgc
     1141 ttggctgacc agcaagatcg atccttctac tggcttcgtc aacctgctca agaccaagta
     1201 caccagcatt gccgacagca agaagttcat cagctcattc gaccggatca tgtacgtgcc
     1261 agaagaggat cttttcgagt tcgccctcga ttacaagaac ttctctagga ccgacgccga
     1321 ctacattaag aagtggaagc tgtactccta cggcaaccgg attcggatct ttcggaaccc
     1381 gaagaaaaac aacgtgttcg actgggaaga ggtaaagttt ccaactttcc tttaccatat
     1441 caaactatag ttcgaaactt tttatttgat caacttcaag gccacccgat ctttctattc
     1501 ctgattaatt tgtgatgaat ccatattgac ttttgatggt tacgcaggtg tgcctgacct
     1561 ctgcctacaa agaactgttc aacaagtacg gcatcaacta ccagcagggt gatattaggg
     1621 ctctgctttg cgagcagtct gacaaggctt tctacagctc tttcatggcc ctgatgtctc
     1681 tgatgctgca aatgaggaac tctatcaccg gtaggaccga tgtggacttc cttatctctc
     1741 cggtgaagaa cagtgacggg atcttctacg acagccggaa ttatgaggct caagagaacg
     1801 caatcctgcc gaagaatgct gatgctaacg gcgcttacaa cattgccaga aaggtgctgt
     1861 gggctatcgg ccagtttaag aaagccgaag atgagaagtt ggacaaggtc tgtctttcct
     1921 atttcatatg tttaatccta ggaatttgat caattgattg tatgtatgtc gatcccaaga
     1981 ctttcttgtt cacttatatc ttaactctct ctttgctgtt tcttgcaggt gaagatcgct
     2041 atctccaaca aagagtggct cgagtacgct cagactagcg ttaagcataa aaggccggcg
     2101 gccacgaaaa aggccggcca ggcaaaaaag aaaaagggtt cgtgagacca cgaagtgccg
     2161 aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta
     2221 tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg
     2281 cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca
     2341 agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta
     2401 cgcagcaggg cagtcgccct aaaacaaagt taaacatcat gggggaagcg gtgatcgccg
     2461 aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt
     2521 tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagcgata
     2581 ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca
     2641 acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag
     2701 tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc
     2761 aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg
     2821 acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc
     2881 cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg
     2941 aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc
     3001 ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg
     3061 ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta
     3121 gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag
     3181 aatttgtcca ttacgtgaaa ggcgagatca ccaaggtagt cggcaaataa tgtctagcta
     3241 gaaattcgtt caagccgacg ccgcttcgcg gcgcggctta actcaagcgt tagatgcact
     3301 aagcacataa ttgctcacag ccaaactatc aggtcaagtc tgcttttatt atttttaagc
     3361 gtgcataata agccctacac aaattgggag atatatcatg ctgtcagacc aagtttactc
     3421 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3481 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3541 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3601 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3661 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3721 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3781 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3841 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3901 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3961 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4021 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4081 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4141 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tgcactctgt
     4201 ggtctcaaat gctgcagcct aagaagaaga gaaaggttgg aggagtcgac tcgagtgcgg
     4261 ccgccacaat gagcaagctc gagaagttta ccaactgcta cagcctgtct aagaccctga
     4321 ggttcaaggc tattcctgtg ggtaagaccc aagagaatat cgacaacaag cggctgctgg
     4381 ttgaggatga gaagagagct gaggattaca agggcgtgaa gaagctgctg gatcggtact
     4441 acctgagctt catcaacgat gtgctgcaca gcatcaagct gaagaacctg aacaactaca
     4501 tcagcctgtt ccggaagaaa acccggaccg agaaagagaa caaagagctt gagaacctcg
     4561 agatcaacct gcggaaagag atcgctaagg ctttcaaggg taacgaaggt aaggattttt
     4621 atgatatact atgcttatgt attttgtact gaaagcatat cctgcttcat tgggatatta
     4681 ctgaaagcat ttaactacat gtaaactcac ttgatgatca ataaacttga ttttgcaggt
     4741 tacaagagcc tgttcaagaa ggatattatc gagactatcc tgcctgagtt cctggacgat
     4801 aaggatgaga ttgccctggt gaacagcttc aacggtttca ctactgcctt caccggtttc
     4861 ttcagaaacc gggaaaacat gttcagcgaa gaggccaagt ctacctctat cgctttccgg
     4921 tgcattaacg agaacttgac ccggtacatc agcaacatgg acatcttcga gaaggtggac
     4981 gccatcttcg ataagcacga ggtgcaagaa atcaaagaga agatcctgaa ctccgactac
     5041 gacgtcgagg atttttttga gggcgagttc ttcaacttcg tgctcaccca agaaggtaac
     5101 attccttagt tacctttctt ttctttttcc atcataagtt tatagattgt acatgctttg
     5161 agatttttct ttgcaaacaa tctcaggtat cgatgtgtac aacgctatca tcggtggttt
     5221 cgtgactgag agcggtgaga agattaaggg cctgaacgag tacattaacc tgtacaatca
     5281 aaagaccaag cagaagctgc cgaagttcaa gccgctttac aagcaggttc tgagcgatcg
     5341 tgagagcctg tctttttacg gagagggata cacctctgat gaagaggttt tggaggtaat
     5401 cttgaaattg gaactcttct tttgttgtct aaacctatca atttctttgc ggaaatttat
     5461 ttgaagctgt agagttaaaa ttgagtcttt taaacttttg taggtgttcc gtaacaccct
     5521 gaacaagaac agcgagatct tcagctccat caagaagctg gaaaagctgt ttaagaactt
     5581 cgacgagtac agcagcgctg gcatcttcgt taagaacggt cctgctatca gcaccatcag
     5641 caaggatatt ttcggcgagt ggaacgtgat ccgggataag tggaatgctg agtacgatga
     5701 catccacctg aagaaaaagg ctgtggtgac cgagaagtac gaggatgata ggcggaagtc
     5761 cttcaagaag ataggtaagt tgttacttat gattgttttc ctctctgcta catgtatttt
     5821 gttgttcatt tctgtaagat ataagaattg agttttcctc tgatgatatt attaggttcc
     5881 tttagcctcg agcagcttca agagtatgct gacgctgatc tgtccgtggt cgagaagctt
     5941 aaagagatca tcatccagaa ggtcgacgag atctacaagg tgtacggcag ctctgagaag
     6001 cttttcgatg ctgacttcgt gttggagaag tctctgaaga agaacgacgc cgttgtcgct
     6061 atcatgaagg atctgctgga cagcgtgaag tctttcgaga actatatcaa ggccttcttc
     6121 ggcgaaggtt agtatcatat gaagaaatac ctagtttcag ttgatgaatg ctattttctg
     6181 acctcagttg ttctcttttg agaattattt cttttctaat ttgcctgatt tttctattaa
     6241 ttcattaggt aaagagacta atagggacga gtcattctac ggcgatttcg tgctggctta
     6301 cgacatcctt cttaaggtgg accacatcta cgacgccatc agaaattacg tgacccagaa
     6361 gccgtacagc aaggacaagt tcaagttgta cttccagaat ccgcagttca tgggcggctg
     6421 ggacaaagac aaagagacag attacagggc taccatcctg cggtacggct ctaagtacta
     6481 ccttgccatc atggacaaga aatacgccaa gtgcctgcaa aagatcgaca aggatgatgt
     6541 gaacggcaac tacgagaaga tcaactacaa gctcctgcca ggtaaatcct ggtccacact
     6601 tttacgataa aaacacaaga ttttaaacta tgaactgatc aataatcatt cctaaaagac
     6661 cacacttttg ttttgtttct aaagtaattt ttactgttat aacaggtcct aacaagatgc
     6721 ttcctaaggt gttcttctca aagaaatgga tggcctacta caacccgagc gaggacatcc
     6781 agaaaatcta caagaacggc accttcaaaa agggcgacat gttcaacctg aacgactgcc
     6841 acaagctgat cgatttcttc aaggacagca tcagccggta tccgaagtgg tctaacgctt
     6901 acgatttcaa cttcagcgag actgagaagt ataaggatat cgccggcttc taccgtgagg
     6961 ttgaggaaca gggttacaag gttagcttcg agagcgccag caagaaagag gtggacaagt
     7021 tggttgaaga aggtaagagg tcaaaaggtt tcc
//
