LOCUS       sequence-322087-        2329 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LbCas12a direct repeat sequence fused to the A. thaliana U6-26 
            promoter used for the assembly of gRNA expression cassettes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2329)
  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 2329)
  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 AtU6-26:LbCas12a_DR (GB1443)
FEATURES             Location/Qualifiers
     source          1..2329
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     178..197
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     425..443
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(447..490)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    493..514
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      795..852
                     /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(876..893)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1047..1635)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1127..1146)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1818..1912)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(1933..2329,1..263))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 acactatccc atatcaccag ctcaccgtct ttcattgcca tacgaaattc cggatgagca
       61 ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt
      121 acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca
      181 actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta
      241 tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca
      301 aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc
      361 ccgatcaact cgagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
      421 taaaaatagg cgtatcacga ggcagaattt cagataaaaa aaatccttag ctttcgctaa
      481 ggatgatttc tggaattcgc ggccgcttct agagcgatga gtggtctcgg gagcatcttc
      541 attcttaaga tatgaagata atcttcaaaa ggcccctggg aatctgaaag aagagaagca
      601 ggcccattta tatgggaaag aacaatagta tttcttatat aggcccattt aagttgaaaa
      661 caatcttcaa aagtcccaca tcgcttagat aagaaaacga agctgagttt atatacagct
      721 agagtcgaag tagtggtaat ttctactaag tgtagattga gaccactcat cgcgactagt
      781 agcggccgct gcagtccggc aaaaaagggc aaggtgtcac caccctgccc tttttcttta
      841 aaaccgaaaa gattacttcg cgttatgcag gcttcctcgc tcactgactc gctgcgctcg
      901 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
      961 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     1021 cgtaaaaagg ccgcgttgct ggcgtttttc cacaggctcc gcccccctga cgagcatcac
     1081 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1141 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1201 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1261 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1321 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1381 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1441 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     1501 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1561 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     1621 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     1681 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     1741 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     1801 gacagctcga ggcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg
     1861 aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag
     1921 ctcgatatca aattacgccc cgccctgcca ctcgtcgcag tactgttgta attcattaag
     1981 cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat
     2041 cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt
     2101 gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgatac
     2161 gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc
     2221 cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag
     2281 tgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtga
//
