LOCUS       sequence-322123-        2328 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AsCas12a 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 2328)
  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 2328)
  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:AsCas12a_DR (GB1442)
FEATURES             Location/Qualifiers
     source          1..2328
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     363..382
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     616..633
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(657..714)
                     /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."
     misc_feature    994..1015
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      1018..1061
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(1065..1083)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             1245..1904
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1311..1330)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1925..2019
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      join(2202..2328,1..462)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
       61 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
      121 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
      181 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
      241 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
      301 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
      361 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
      421 cgatttttgt gatgctcgtc aggggggcgg agcctgtgga aaaacgccag caacgcggcc
      481 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
      541 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
      601 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcct gcataacgcg aagtaatctt
      661 ttcggtttta aagaaaaagg gcagggtggt gacaccttgc ccttttttgc cggactgcag
      721 cggccgctac tagtcgcgat gagtggtctc aatctacaag agtagaaatt accactactt
      781 cgactctagc tgtatataaa ctcagcttcg ttttcttatc taagcgatgt gggacttttg
      841 aagattgttt tcaacttaaa tgggcctata taagaaatac tattgttctt tcccatataa
      901 atgggcctgc ttctcttctt tcagattccc aggggccttt tgaagattat cttcatatct
      961 taagaatgaa gatgctcccg agaccactca tcgctctaga agcggccgcg aattccagaa
     1021 atcatcctta gcgaaagcta aggatttttt ttatctgaaa ttctgcctcg tgatacgcct
     1081 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcactcgagt
     1141 tgatcgggca cgtaagaggt tccaactttc accataatga aataagatca ctaccgggcg
     1201 tattttttga gttatcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca
     1261 ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc
     1321 agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa
     1381 agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc
     1441 tgatgaatgc tcatccggaa tttcgtatgg caatgaaaga cggtgagctg gtgatatggg
     1501 atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcactct
     1561 ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt
     1621 gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtat
     1681 cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact
     1741 tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc
     1801 cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta
     1861 atgaattaca acagtactgc gacgagtggc agggcggggc gtaatttgat atcgagctcg
     1921 cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga
     1981 acgctcggtt gccgccgggc gttttttatt ggtgagaatc caagcctcga gctgtcagac
     2041 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2101 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2161 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2221 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2281 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcaga
//
