LOCUS       sequence-176062-        4991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LbCpf1 Gateway gRNA entry plasmid using Zea mays Ubi promoter and 
            ribozyme processing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4991)
  AUTHORS   Tang X, Lowder LG, Zhang T, Malzahn AA, Zheng X, Voytas DF, Zhong Z,
            Chen Y, Ren Q, Li Q, Kirkland ER, Zhang Y, Qi Y
  TITLE     A CRISPR-Cpf1 system for efficient genome editing and 
            transcriptional repression in plants.
  JOURNAL   Nat Plants. 2017 Feb 17;3:17018. doi: 10.1038/nplants.2017.18.
  PUBMED    28211909
REFERENCE   2  (bases 1 to 4991)
  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: "Nat 
            Plants."; date: "2017-02-17"; volume: "3:17018. doi"; pages: " 
            10.1038/nplants.2017.18"
COMMENT     Sequence Label: pYPQ141-ZmUbi-RZ-Lb
FEATURES             Location/Qualifiers
     source          1..4991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     ncRNA           1040..1076
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     ncRNA           1130..1197
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     protein_bind    complement(1217..1316)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1334..1352)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1369..1387)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1392..1408)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1392..1408)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1839..2630
                     /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      2723..3311
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3212..3231
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3465..3482
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3641..3668)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3760..3846)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3895..3917
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3909..3926
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3910..3926
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3941..4036
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 cctccgcttc aaggtacgcc gctcgtcctc cccccccccc cctctctacc ttctctagat
       61 cggcgttccg gtccatggtt agggcccggt agttctactt ctgttcatgt ttgtgttaga
      121 tccgtgtttg tgttagatcc gtgctgctag cgttcgtaca cggatgcgac ctgtacgtca
      181 gacacgttct gattgctaac ttgccagtgt ttctctttgg ggaatcctgg gatggctcta
      241 gccgttccgc agacgggatc gatttcatga ttttttttgt ttcgttgcat agggtttggt
      301 ttgccctttt cctttatttc aatatatgcc gtgcacttgt ttgtcgggtc atcttttcat
      361 gctttttttt gtcttggttg tgatgatgtg gtctggttgg gcggtcgttc tagatcggag
      421 tagaattaat tctgtttcaa actacctggt ggatttatta attttggatc tgtatgtgtg
      481 tgccatacat attcatagtt acgaattgaa gatgatggat ggaaatatcg atctaggata
      541 ggtatacatg ttgatgcggg ttttactgat gcatatacag agatgctttt tgttcgcttg
      601 gttgtgatga tgtggtgtgg ttgggcggtc gttcattcgt tctagatcgg agtagaatac
      661 tgtttcaaac tacctggtgt atttattaat tttggaactg tatgtgtgtg tcatacatct
      721 tcatagttac gagtttaaga tggatggaaa tatcgatcta ggataggtat acatgttgat
      781 gtgggtttta ctgatgcata tacatgatgg catatgcagc atctattcat atgctctaac
      841 cttgagtacc tatctattat aataaacaag tatgttttat aattattttg atcttgatat
      901 acttggatga tggcatatgc agcagctata tgtggatttt tttagccctg ccttcatacg
      961 ctatttattt gcttggtact gtttcttttg tcgatgctca ccctgttgtt tggtgttact
     1021 tctgcaggga tccaaattac tgatgagtcc gtgaggacga aacgagtaag ctcgtctaat
     1081 ttctactaag tgtagatgag acggagctca gtctgaccgc ggcgtctctg gccggcatgg
     1141 tcccagcctc ctcgctggcg ccggctgggc aacatgcttc ggcatggcga atgggacggt
     1201 acccggccgg aattcgaccc agctttcttg tacaaagttg gcattataaa aaataattgc
     1261 tcatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat
     1321 ccagctgata tcccctatag tgagtcgtat tacatggtca tagcatcccc tatagtgagt
     1381 cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat
     1441 gttacattgc acaagataaa aatatatcat catgcctcct ctggaccagc caggacagaa
     1501 atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg aaacggatga
     1561 aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta
     1621 tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg
     1681 cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca
     1741 agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta
     1801 cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg gtgatcgccg
     1861 aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt
     1921 tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata
     1981 ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca
     2041 acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag
     2101 tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc
     2161 aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg
     2221 acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc
     2281 cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg
     2341 aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc
     2401 ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg
     2461 ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta
     2521 gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag
     2581 aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc
     2641 acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg
     2701 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2761 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2821 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2881 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2941 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3001 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3061 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3121 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3181 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3241 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3301 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3361 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3421 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3481 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     3541 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     3601 atacgcgtac cgcgagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat
     3661 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc
     3721 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc
     3781 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt
     3841 tatttgatgc ctggcagttc cctactctcg cgttaacgct tgcatggatg ttttcccagt
     3901 cacgacgttg taaaacgacg gccagtctta agctcgggcc caataatgat tttattttga
     3961 ctgatagtga cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact
     4021 ttgtatacaa aagttgcccc atggcgttcc ctctagagag ataatgagca ttgcatgtct
     4081 aagttataaa aaattaccac atattttttt tgtcacactt gtttgaagtg cagtttatct
     4141 atctttatac atatatttaa actttactct acgaataata taatctatag tactacaata
     4201 atatcagtgt tttagagaat catataaatg aacagttaga catggtctaa aggacaattg
     4261 agtattttga caacaggact ctacagtttt atctttttag tgtgcatgtg ttctcctttt
     4321 tttttgcaaa tagcttcacc tatataatac ttcatccatt ttattagtac atccatttag
     4381 ggtttagggt taatggtttt tatagactaa tttttttagt acatctattt tattctattt
     4441 tagcctctaa attaagaaaa ctaaaactct attttagttt ttttatttaa taatttagat
     4501 ataaaataga ataaaataaa gtgactaaaa attaaacaaa taccctttaa gaaattaaaa
     4561 aaactaagga aacatttttc ttgtttcgag tagataatgc cagcctgtta aacgccgtcg
     4621 acgagtctaa cggacaccaa ccagcgaacc agcagcgtcg cgtcgggcca agcgaagcag
     4681 acggcacggc atctctgtcg ctgcctctgg acccctctcg agagttccgc tccaccgttg
     4741 gacttgctcc gctgtcggca tccagaaatt gcgtggcgga gcggcagacg tgagccggca
     4801 cggcaggcgg cctcctcctc ctctcacggc accggcagct acgggggatt cctttcccac
     4861 cgctccttcg ctttcccttc ctcgcccgcc gtaataaata gacaccccct ccacaccctc
     4921 tttccccaac ctcgtgttgt tcggagcgca cacacacaca accagatctc ccccaaatcc
     4981 acccgtcggc a
//
