LOCUS Exported 7008 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway entry plasmid (attL1. ACCESSION . VERSION . KEYWORDS pYPQ284-RVRR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7008) AUTHORS Zhang Y, Ren Q, Tang X, Liu S, Malzahn AA, Zhou J, Wang J, Yin D, Pan C, Yuan M, Huang L, Yang H, Zhao Y, Fang Q, Zheng X, Tian L, Cheng Y, Le Y, McCoy B, Franklin L, Selengut JD, Mount SM, Que Q, Zhang Y, Qi Y TITLE Expanding the scope of plant genome engineering with Cas12a orthologs and highly multiplexable editing systems. JOURNAL Nat Commun. 2021 Mar 29;12(1):1944. doi: 10.1038/s41467-021-22330-w. PUBMED 33782402 REFERENCE 2 (bases 1 to 7008) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7008 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 358..1149 /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 1242..1830 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1731..1750 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1984..2001 /label=L4440 /note="L4440 vector, forward primer" terminator 2160..2187 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 2279..2365 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 2414..2436 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2428..2445 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2429..2445 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 2461..2560 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 2585..2605 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 6380..6427 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" terminator 6458..6710 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" protein_bind 6755..6878 /gene="mutant version of attR" /label=attR5 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(6895..6914) /label=T7 /note="T7 promoter, forward primer" promoter complement(6896..6914) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6919..6935) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6919..6935) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 tggaccagcc aggacagaaa tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc 61 acaccgtgga aacggatgaa ggcacgaacc cagtggacat aagcctgttc ggttcgtaag 121 ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc 181 ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt tggggtacag 241 tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta 301 tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt aaacatcatg 361 agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag 421 cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc 481 ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca 541 acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagcgag 601 attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat 661 ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct tgcaggtatc 721 ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat 781 agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat 841 ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggc 901 gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa 961 atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc ccagtatcag 1021 cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg cttggcctcg 1081 cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc 1141 ggcaaataac cctcgagcca cccatgacca aaatccctta acgtgagtta cgcgtcgttc 1201 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 1261 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 1321 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 1381 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 1441 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 1501 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 1561 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 1621 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 1681 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 1741 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 1801 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 1861 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg 1921 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag 1981 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc 2041 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc 2101 agtgagcgca acgcaattaa tacgcgtacc gcgagccagg aagagtttgt agaaacgcaa 2161 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc 2221 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat 2281 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt 2341 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgctt 2401 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc 2461 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa 2521 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccgaattcgc ccttcaccat 2581 ggctcctaag aagaagcgga aggttggtat tcacggggtg cctgcggctc tgtttcaaga 2641 ttttacacat ctgtacccgc tgagtaaaac agtgcggttc gagctgaaac ccataggaag 2701 gaccctcgag cacatccacg cgaagaattt tctgagccag gatgaaacta tggctgatat 2761 gtatcaaaaa gttaaggtca ttttggacga ctatcatcgc gattttattg ccgacatgat 2821 gggagaggtg aaactcacga agcttgctga attttacgac gtctatctga agttcaggaa 2881 aaatcctaag gacgatgggc tgcaaaaaca gcttaaagac cttcaagctg tccttcggaa 2941 ggaatcggtg aagcctatag ggtcaggtgg gaagtacaaa acaggctacg atagactctt 3001 tggggcaaaa ctcttcaaag atggaaaaga gttgggtgac ctcgcaaaat tcgttatagc 3061 ccaagaaggt gagtcttctc cgaagctggc tcatcttgct cattttgaga agttcagcac 3121 gtattttact ggatttcacg ataatcggaa gaatatgtac tcggatgaag acaagcatac 3181 tgcaatagcg tacaggctca tccatgagaa tttgccgaga ttcatcgaca atctgcaaat 3241 cttgacaaca atcaaacaaa agcatagcgc cctctatgat cagataatca acgagctcac 3301 ggcctccggg ctcgacgtct ccttggcttc tcatcttgac gggtatcaca agctccttac 3361 acaagagggg atcacggcat acaacaggat cataggagag gtgaatggat atacaaataa 3421 gcataaccag atatgccaca agagcgagcg catagcgaaa cttagaccct tgcacaagca 3481 aatcctttct gacggaatgg gagtgtcatt ccttccgtct aagttcgcgg atgatagtga 3541 gatgtgccaa gcggtcaacg aattttatcg ccattatact gacgtgttcg caaaggtgca 3601 aagtctcttt gacggatttg atgatcacca gaaagacggg atctatgttg aacacaaaaa 3661 ccttaatgaa ctgagcaaac aggcgttcgg cgactttgct ttgctgggga gggtccttga 3721 tggatactac gtggacgttg tcaatccgga gttcaatgag cggttcgcaa aggccaagac 3781 tgacaatgcg aaagccaagc ttacaaaaga aaaggacaaa ttcattaaag gagtccactc 3841 actggcttcc ctcgaacaag caatagaaca ccatacagct agacacgacg atgagagtgt 3901 tcaagccgga aaacttggcc agtacttcaa acacggtttg gcgggggttg acaacccgat 3961 tcagaaaatt cacaataacc attcgacgat taaagggttt ctggaaaggg aaaggcctgc 4021 tggggaacgg gcgctcccga agatcaagtc aggaaaaaac ccagaaatga cacagctcag 4081 gcagctgaag gaacttttgg acaacgcatt gaatgtggcg cacttcgcta agctgctgac 4141 aactaaaaca accttggaca accaggatgg aaatttttac ggggagtttg gggtgcttta 4201 cgacgagctg gctaaaattc caactctcta caataaggtt agagattatc tctctcaaaa 4261 gcccttttct accgaaaagt ataagctcaa cttcggcaat ccgacccttc tccgcgggtg 4321 ggacctgaac gtggagaaag atcgctttgg ggttatactt cagaaggatg gatgctatta 4381 cttggcgctt cttgataagg ctcataaaaa agttttcgac aacgccccta acactggtaa 4441 gaacgtctac caaaagatgg tctacaaact gttgcccggc cccaacaaaa tgcttcctag 4501 ggtgtttttc gcaaaatcga atctcgacta ttataatcca tctgccgagc tccttgacaa 4561 atatgctaag gggacccata aaaagggtga taatttcaac ctgaaggact gccacgcgct 4621 tatcgacttt ttcaaagccg ggataaataa gcatccggag tggcaacatt ttggttttaa 4681 attttcgcca acgtcgtcct atcgcgacct ttccgatttc tatagggaag ttgaacctca 4741 ggggtaccag gtcaaatttg ttgacattaa tgcggactac attgatgaat tggtggagca 4801 agggaagctc tacctctttc aaatatataa caaagatttc tcgccaaaag cgcatggtaa 4861 accgaatctt cataccttgt actttaaagc acttttttca gaagataact tggcggaccc 4921 gatctacaag ctgaatgggg aagctcagat cttctacagg aaagcttcgt tggacatgaa 4981 cgagactacc atacatcgcg cgggagaggt gcttgagaac aaaaatcccg acaacccgaa 5041 aaagcggcaa ttcgtttacg acatcatcaa agacaaacgg tacacgcagg acaaatttat 5101 gctccacgtc cccattacca tgaattttgg agtccaaggc atgaccatta aggaattcaa 5161 caaaaaggtc aaccaaagta ttcagcaata cgatgaagtc aatgtcatag gcatagatcg 5221 gggagaaagg catctgttgt atcttaccgt gattaactct aagggtgaaa tactggagca 5281 acggtcactt aacgatataa ccacggcgtc cgcgaacggt acacaagtga ccactcccta 5341 ccacaaaata ttggataaaa gggagataga acgcttgaat gcccgcgttg gctggggtga 5401 gattgagacc atcaaagagc ttaaatcggg atatttgtct cacgtcgttc atcaaattaa 5461 ccaactcatg cttaagtaca atgcaatcgt tgtgctcgag gacctgaact ttggtttcaa 5521 aagagggagg ttcaaggtgg aaaaacaaat ttaccagaac tttgaaaacg cgcttatcaa 5581 gaaattgaat caccttgttt tgaaagataa ggcagatgac gaaatcgggt cgtataaaaa 5641 tgcactccag ttgacaaata atttcacgga tttgaagtcg atcggcaagc aaacagggtt 5701 cctcttttat gtgccagcgt ggaatacatc aaaaattgat ccggagacgg gatttgtcga 5761 cttgctgaag cctaggtatg agaacattgc ccaatctcag gcctttttcg gcaaattcga 5821 taaaatatgc tacaacacag acaaaggtta ttttgaattt cacattgatt acgccaaatt 5881 tacagataag gcgaaaaaca gcagacagaa atgggctatc tgttctcatg gggacaaacg 5941 ctatgtctac gataagacgg ctaatcaaaa taaaggcgcc gcaaaaggta ttaatgtgaa 6001 tgatgagctg aaaagcttgt ttgcccgcta ccatatcaat gataaacaac caaacttggt 6061 gatggacata tgccagaaca atgacaaaga attccacaag tcactcatgt gcctgcttaa 6121 aacccttttg gcgctgcggt atagcaatgc atctagcgat gaagacttta ttttgagtcc 6181 cgtggccaac gacgagggcg tgttttttaa ttcagccttg gcggacgata cgcagcccca 6241 gaatgcggac gcaaacggcg cgtaccacat tgcactgaag ggactgtggc ttctgaacga 6301 gctgaaaaat agcgacgacc tgaataaagt caagttggcc attgacaatc aaacctggtt 6361 gaatttcgct caaaatagaa agcgtcctgc tgccaccaaa aaggccggac aggctaagaa 6421 aaagaagtga gacgactagt ggcggccgcc gacgtccgat cgttcaaaca tttggcaata 6481 aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt 6541 gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt 6601 ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg 6661 cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc gggaattgat 6721 cccccctcga cagcttccgg aaagggcgaa ttcgcaactt tgtatacaaa agttgaacga 6781 gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact 6841 acataatact gtaaaacaca acatatccag tcactatgcc atccagctga tatcccctat 6901 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat 6961 ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctc //