LOCUS       sequence-324726-        5844 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human codon optimized coding sequence of Cas12a from Lachnospiraceae
            CRISPR system (LbCas12a)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5844)
  AUTHORS   Bernabe-Orts JM, Casas-Rodrigo I, Minguet EG, Landolfi V, 
            Garcia-Carpintero V, Gianoglio S, Vazquez-Vilar M, Granell A, Orzaez
            D
  TITLE     Assessment of Cas12a-mediated gene editing efficiency in plants.
  JOURNAL   Plant Biotechnol J. 2019 Apr 5. doi: 10.1111/pbi.13113.
  PUBMED    30950179
REFERENCE   2  (bases 1 to 5844)
  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 
            Biotechnol J. 2019 Apr 5. doi: 10.1111/pbi.13113."
COMMENT     Sequence Label: pUPD2 LbCas12a (GB1439)
FEATURES             Location/Qualifiers
     source          1..5844
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(167..261)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(282..941)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     856..875
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1103..1121
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1125..1168)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1171..1192
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             1209..4892
                     /codon_start=1
                     /product="CRISPR-associated protein from Lachnospiraceae 
                     bacterium ND2006 (Zetsche et al., 2015)"
                     /label=LbCpf1
                     /translation="MSKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAE
                     DYKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKE
                     IAKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFDNRENMFS
                     EEAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFEG
                     EFFNFVLTQEGIDVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVL
                     SDRESLSFYGEGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKN
                     GPAISTISKDIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLEQ
                     LQEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKD
                     LLDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYS
                     KDKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVNG
                     NYEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKL
                     IDFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLV
                     EEGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLKK
                     EELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKI
                     NTEVRVLLKHDDNPYVIGIDRGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTD
                     YHSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNSG
                     FKNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQ
                     NGFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALD
                     YKNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQ
                     QGDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSRN
                     YEAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTS
                     VKH"
     CDS             4893..4940
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      4988..5045
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(5069..5086)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5240..5828)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5320..5339)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
       61 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
      121 tctaaagtat atatgagtaa acttggtctg acagctcgag gcttggattc tcaccaataa
      181 aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta
      241 ctggatctat caacaggagt ccaagcgagc tcgatatcaa attacgcccc gccctgccac
      301 tcgtcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg
      361 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
      421 cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
      481 gtgaaactca cccagggatt ggctgatacg aaaaacatat tctcaataaa ccctttaggg
      541 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc
      601 cggaaatcgt cgtggtattc actccagagt gatgaaaacg tttcagtttg ctcatggaaa
      661 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
      721 cgaaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac
      781 ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg
      841 ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg
      901 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct
      961 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     1021 ttggaacctc ttacgtgccc gatcaactcg agtgccacct gacgtctaag aaaccattat
     1081 tatcatgaca ttaacctata aaaataggcg tatcacgagg cagaatttca gataaaaaaa
     1141 atccttagct ttcgctaagg atgatttctg gaattcgcgg ccgcttctag agcgatgagt
     1201 ggtctcgaat gagcaagctg gagaagttta caaactgcta ctccctgtct aagaccctga
     1261 ggttcaaggc catccctgtg ggcaagaccc aggagaacat cgacaataag cggctgctgg
     1321 tggaggacga gaagagagcc gaggattata agggcgtgaa gaagctgctg gatcgctact
     1381 atctgtcttt tatcaacgac gtgctgcaca gcatcaagct gaagaatctg aacaattaca
     1441 tcagcctgtt ccggaagaaa accagaaccg agaaggagaa taaggagctg gagaacctgg
     1501 agatcaatct gcggaaggag atcgccaagg ccttcaaggg caacgagggc tacaagtccc
     1561 tgtttaagaa ggatatcatc gagacaatcc tgccagagtt cctggacgat aaggacgaga
     1621 tcgccctggt gaacagcttc aatggcttta ccacagcctt caccggcttc tttgataaca
     1681 gagagaatat gttttccgag gaggccaaga gcacatccat cgccttcagg tgtatcaacg
     1741 agaatctgac ccgctacatc tctaatatgg acatcttcga gaaggtggac gccatctttg
     1801 ataagcacga ggtgcaggag atcaaggaga agatcctgaa cagcgactat gatgtggagg
     1861 atttctttga gggcgagttc tttaactttg tgctgacaca ggagggcatc gacgtgtata
     1921 acgccatcat cggcggcttc gtgaccgaga gcggcgagaa gatcaagggc ctgaacgagt
     1981 acatcaacct gtataatcag aaaaccaagc agaagctgcc taagtttaag ccactgtata
     2041 agcaggtgct gagcgatcgg gagtctctga gcttctacgg cgagggctat acatccgatg
     2101 aggaggtgct ggaggtgttt agaaacaccc tgaacaagaa cagcgagatc ttcagctcca
     2161 tcaagaagct ggagaagctg ttcaagaatt ttgacgagta ctctagcgcc ggcatctttg
     2221 tgaagaacgg ccccgccatc agcacaatct ccaaggatat cttcggcgag tggaacgtga
     2281 tccgggacaa gtggaatgcc gagtatgacg atatccacct gaagaagaag gccgtggtga
     2341 ccgagaagta cgaggacgat cggagaaagt ccttcaagaa gatcggctcc ttttctctgg
     2401 agcagctgca ggagtacgcc gacgccgatc tgtctgtggt ggagaagctg aaggagatca
     2461 tcatccagaa ggtggatgag atctacaagg tgtatggctc ctctgagaag ctgttcgacg
     2521 ccgattttgt gctggagaag agcctgaaga agaacgacgc cgtggtggcc atcatgaagg
     2581 acctgctgga ttctgtgaag agcttcgaga attacatcaa ggccttcttt ggcgagggca
     2641 aggagacaaa cagggacgag tccttctatg gcgattttgt gctggcctac gacatcctgc
     2701 tgaaggtgga ccacatctac gatgccatcc gcaattatgt gacccagaag ccctactcta
     2761 aggataagtt caagctgtat tttcagaacc ctcagttcat gggcggctgg gacaaggata
     2821 aggagacaga ctatcgggcc accatcctga gatacggctc caagtactat ctggccatca
     2881 tggataagaa gtacgccaag tgcctgcaga agatcgacaa ggacgatgtg aacggcaatt
     2941 acgagaagat caactataag ctgctgcccg gccctaataa gatgctgcca aaggtgttct
     3001 tttctaagaa gtggatggcc tactataacc ccagcgagga catccagaag atctacaaga
     3061 atggcacatt caagaagggc gatatgttta acctgaatga ctgtcacaag ctgatcgact
     3121 tctttaagga tagcatctcc cggtatccaa agtggtccaa tgcctacgat ttcaactttt
     3181 ctgagacaga gaagtataag gacatcgccg gcttttacag agaggtggag gagcagggct
     3241 ataaggtgag cttcgagtct gccagcaaga aggaggtgga taagctggtg gaggagggca
     3301 agctgtatat gttccagatc tataacaagg acttttccga taagtctcac ggcacaccca
     3361 atctgcacac catgtacttc aagctgctgt ttgacgagaa caatcacgga cagatcaggc
     3421 tgagcggagg agcagagctg ttcatgaggc gcgcctccct gaagaaggag gagctggtgg
     3481 tgcacccagc caactcccct atcgccaaca agaatccaga taatcccaag aaaaccacaa
     3541 ccctgtccta cgacgtgtat aaggataaga ggttttctga ggaccagtac gagctgcaca
     3601 tcccaatcgc catcaataag tgccccaaga acatcttcaa gatcaataca gaggtgcgcg
     3661 tgctgctgaa gcacgacgat aacccctatg tgatcggcat cgataggggc gagcgcaatc
     3721 tgctgtatat cgtggtggtg gacggcaagg gcaacatcgt ggagcagtat tccctgaacg
     3781 agatcatcaa caacttcaac ggcatcagga tcaagacaga ttaccactct ctgctggaca
     3841 agaaggagaa ggagaggttc gaggcccgcc agaactggac ctccatcgag aatatcaagg
     3901 agctgaaggc cggctatatc tctcaggtgg tgcacaagat ctgcgagctg gtggagaagt
     3961 acgatgccgt gatcgccctg gaggacctga actctggctt taagaatagc cgcgtgaagg
     4021 tggagaagca ggtgtatcag aagttcgaga agatgctgat cgataagctg aactacatgg
     4081 tggacaagaa gtctaatcct tgtgcaacag gcggcgccct gaagggctat cagatcacca
     4141 ataagttcga gagctttaag tccatgtcta cccagaacgg cttcatcttt tacatccctg
     4201 cctggctgac atccaagatc gatccatcta ccggctttgt gaacctgctg aaaaccaagt
     4261 ataccagcat cgccgattcc aagaagttca tcagctcctt tgacaggatc atgtacgtgc
     4321 ccgaggagga tctgttcgag tttgccctgg actataagaa cttctctcgc acagacgccg
     4381 attacatcaa gaagtggaag ctgtactcct acggcaaccg gatcagaatc ttccggaatc
     4441 ctaagaagaa caacgtgttc gactgggagg aggtgtgcct gaccagcgcc tataaggagc
     4501 tgttcaacaa gtacggcatc aattatcagc agggcgatat cagagccctg ctgtgcgagc
     4561 agtccgacaa ggccttctac tctagcttta tggccctgat gagcctgatg ctgcagatgc
     4621 ggaacagcat cacaggccgc accgacgtgg attttctgat cagccctgtg aagaactccg
     4681 acggcatctt ctacgatagc cggaactatg aggcccagga gaatgccatc ctgccaaaga
     4741 acgccgacgc caatggcgcc tataacatcg ccagaaaggt gctgtgggcc atcggccagt
     4801 tcaagaaggc cgaggacgag aagctggata aggtgaagat cgccatctct aacaaggagt
     4861 ggctggagta cgcccagacc agcgtgaagc acaaaaggcc ggcggccacg aaaaaggccg
     4921 gccaggcaaa aaagaaaaag tagtgagctt tgagaccact catcgcgact agtagcggcc
     4981 gctgcagtcc ggcaaaaaag ggcaaggtgt caccaccctg ccctttttct ttaaaaccga
     5041 aaagattact tcgcgttatg caggcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5101 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5161 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5221 aggccgcgtt gctggcgttt ttccacaggc tccgcccccc tgacgagcat cacaaaaatc
     5281 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5341 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5401 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5461 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5521 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5581 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5641 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5701 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5761 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5821 gatctcaaga agatcctttg atct
//
