LOCUS Exported 3043 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Level 0 Golden Gate Part: Coding sequence, neomycin phophotransferase II (Escherichia coli). ACCESSION . VERSION . KEYWORDS pICSL80037 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3043) AUTHORS Lawrenson T, Shorinola O, Stacey N, Li C, Ostergaard L, Patron N, Uauy C, Harwood W TITLE Induction of targeted, heritable mutations in barley and Brassica oleracea using RNA-guided Cas9 nuclease. JOURNAL Genome Biol. 2015 Nov 30;16(1):258. doi: 10.1186/s13059-015-0826-7. PUBMED 26616834 REFERENCE 2 (bases 1 to 3043) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3043 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(40..58) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 126..230 /gene="bla" /label=AmpR promoter CDS 450..1241 /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 1412..2000 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1901..1920 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2154..2171 /label=L4440 /note="L4440 vector, forward primer" CDS 2249..3043 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2303..2322) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2913..2932 /label=Neo-F /note="Neomycin resistance gene, forward primer" ORIGIN 1 gctttgagac cacgaagtgg ctcttcagtg gacgaaaggg cctcgtgata cgcctatttt 61 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa 121 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca 181 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgcgctcac 241 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca 301 tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag 361 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg 421 gcagtcgccc taaaacaaag ttaaacatca tgagggaagc ggtgatcgcc gaagtatcga 481 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg 541 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagcgat attgatttgc 601 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt 661 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg 721 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag 781 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc 841 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg 901 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa 961 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt 1021 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact 1081 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt 1141 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc 1201 attacgtaaa aggcgagatc accaaggtag tcggcaaata actgtcagac caagtttact 1261 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga 1321 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt 1381 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct 1441 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 1501 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc 1561 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 1621 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 1681 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 1741 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 1801 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 1861 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 1921 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 1981 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt 2041 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta 2101 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt 2161 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc 2221 cgattcatta atcactctgt ggtctcaaat gattgaacaa gatggattgc acgcaggttc 2281 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg 2341 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac 2401 cgacctgtcc ggtgccctga atgaactcca agacgaggca gcgcggctat cgtggctggc 2461 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg 2521 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga 2581 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg 2641 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg 2701 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt 2761 cgccaggctc aaggcgcgga tgcccgacgg cgaggatctc gtcgtgaccc acggcgatgc 2821 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg 2881 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga 2941 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc 3001 gcagcgcatc gccttctatc gccttcttga cgagttcttc tga //