LOCUS Exported 4991 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AsCpf1 gRNA entry plasmid using Zea mays Ubi promoter and ribozyme processing. ACCESSION . VERSION . KEYWORDS pYPQ141-ZmUbi-RZ-As 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4991 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 4..26 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 18..35 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 19..35 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 50..145 /gene="mutant version of attL" /label=attL5 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(2317..2416) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter complement(2434..2452) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(2469..2487) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2492..2508) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2492..2508) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 2939..3730 /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 3823..4411 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4312..4331 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4565..4582 /label=L4440 /note="L4440 vector, forward primer" terminator 4741..4768 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 4860..4946 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" ORIGIN 1 tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc aataatgatt 61 ttattttgac tgatagtgac ctgttcgttg caacaaattg atgagcaatg cttttttata 121 atgccaactt tgtatacaaa agttgcccca tggcgttccc tctagagaga taatgagcat 181 tgcatgtcta agttataaaa aattaccaca tatttttttt gtcacacttg tttgaagtgc 241 agtttatcta tctttataca tatatttaaa ctttactcta cgaataatat aatctatagt 301 actacaataa tatcagtgtt ttagagaatc atataaatga acagttagac atggtctaaa 361 ggacaattga gtattttgac aacaggactc tacagtttta tctttttagt gtgcatgtgt 421 tctccttttt ttttgcaaat agcttcacct atataatact tcatccattt tattagtaca 481 tccatttagg gtttagggtt aatggttttt atagactaat ttttttagta catctatttt 541 attctatttt agcctctaaa ttaagaaaac taaaactcta ttttagtttt tttatttaat 601 aatttagata taaaatagaa taaaataaag tgactaaaaa ttaaacaaat accctttaag 661 aaattaaaaa aactaaggaa acatttttct tgtttcgagt agataatgcc agcctgttaa 721 acgccgtcga cgagtctaac ggacaccaac cagcgaacca gcagcgtcgc gtcgggccaa 781 gcgaagcaga cggcacggca tctctgtcgc tgcctctgga cccctctcga gagttccgct 841 ccaccgttgg acttgctccg ctgtcggcat ccagaaattg cgtggcggag cggcagacgt 901 gagccggcac ggcaggcggc ctcctcctcc tctcacggca ccggcagcta cgggggattc 961 ctttcccacc gctccttcgc tttcccttcc tcgcccgccg taataaatag acaccccctc 1021 cacaccctct ttccccaacc tcgtgttgtt cggagcgcac acacacacaa ccagatctcc 1081 cccaaatcca cccgtcggca cctccgcttc aaggtacgcc gctcgtcctc cccccccccc 1141 ccctctctac cttctctaga tcggcgttcc ggtccatggt tagggcccgg tagttctact 1201 tctgttcatg tttgtgttag atccgtgttt gtgttagatc cgtgctgcta gcgttcgtac 1261 acggatgcga cctgtacgtc agacacgttc tgattgctaa cttgccagtg tttctctttg 1321 gggaatcctg ggatggctct agccgttccg cagacgggat cgatttcatg attttttttg 1381 tttcgttgca tagggtttgg tttgcccttt tcctttattt caatatatgc cgtgcacttg 1441 tttgtcgggt catcttttca tgcttttttt tgtcttggtt gtgatgatgt ggtctggttg 1501 ggcggtcgtt ctagatcgga gtagaattaa ttctgtttca aactacctgg tggatttatt 1561 aattttggat ctgtatgtgt gtgccataca tattcatagt tacgaattga agatgatgga 1621 tggaaatatc gatctaggat aggtatacat gttgatgcgg gttttactga tgcatataca 1681 gagatgcttt ttgttcgctt ggttgtgatg atgtggtgtg gttgggcggt cgttcattcg 1741 ttctagatcg gagtagaata ctgtttcaaa ctacctggtg tatttattaa ttttggaact 1801 gtatgtgtgt gtcatacatc ttcatagtta cgagtttaag atggatggaa atatcgatct 1861 aggataggta tacatgttga tgtgggtttt actgatgcat atacatgatg gcatatgcag 1921 catctattca tatgctctaa ccttgagtac ctatctatta taataaacaa gtatgtttta 1981 taattatttt gatcttgata tacttggatg atggcatatg cagcagctat atgtggattt 2041 ttttagccct gccttcatac gctatttatt tgcttggtac tgtttctttt gtcgatgctc 2101 accctgttgt ttggtgttac ttctgcaggg atccaaatta ctgatgagtc cgtgaggacg 2161 aaacgagtaa gctcgtctaa tttctactct tgtagatgag acggagctca gtctgaccgc 2221 ggcgtctctg gccggcatgg tcccagcctc ctcgctggcg ccggctgggc aacatgcttc 2281 ggcatggcga atgggacggt acccggccgg aattcgaccc agctttcttg tacaaagttg 2341 gcattataaa aaataattgc tcatcaattt gttgcaacga acaggtcact atcagtcaaa 2401 ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca 2461 tagcatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc 2521 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgcctcct 2581 ctggaccagc caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg 2641 cacaccgtgg aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa 2701 gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag 2761 cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca 2821 gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt 2881 atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat 2941 gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga 3001 gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg 3061 cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac 3121 aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga 3181 gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta 3241 tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat 3301 cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca 3361 tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga 3421 tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg 3481 cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa 3541 aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca 3601 gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc 3661 gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt 3721 cggcaaataa ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt 3781 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct 3841 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc 3901 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc 3961 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc 4021 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc 4081 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg 4141 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata 4201 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta 4261 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc 4321 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg 4381 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt 4441 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt 4501 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga 4561 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc 4621 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg 4681 cagtgagcgc aacgcaatta atacgcgtac cgcgagccag gaagagtttg tagaaacgca 4741 aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg 4801 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga 4861 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct 4921 tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct 4981 tgcatggatg t //