LOCUS Exported 5188 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION P3b. ACCESSION . VERSION . KEYWORDS pMR-A-4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5188) AUTHORS Richter MF, Zhao KT, Eton E, Lapinaite A, Newby GA, Thuronyi BW, Wilson C, Koblan LW, Zeng J, Bauer DE, Doudna JA, Liu DR TITLE Phage-assisted evolution of an adenine base editor with improved Cas domain compatibility and activity JOURNAL Nat Biotechnol (2020) REFERENCE 2 (bases 1 to 5188) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5188 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 301..320 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 554..571 /label=L4440 /note="L4440 vector, forward primer" misc_feature 586..726 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(652..671) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 812..834 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(828..1019) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4109..4900) /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" promoter complement(4901..4972) /gene="bla" /label=AmpR promoter rep_origin 5000..400 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 61 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 121 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga 181 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc 241 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 301 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg 361 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt 421 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc 481 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg 541 aacgaccgag cgcagcgagt cagtgagcga ggaagcggat gagcgcctga tgcggtattt 601 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg 661 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg 721 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg 781 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca 841 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc 901 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt 961 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact 1021 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag 1081 aggatgctca cgatacgggt tactgatgat gaacaagagg acatccggtc aaataaaacg 1141 aaaggctcag tcgaaagact gggcctttcg ttttgctgag gagacttagg gactctcaca 1201 gctaacacca cgtcgtccct atctgctgcc ctaggtctat gagtggttgc tggataactt 1261 tacgggcatg cataaggctc gtatgatata ttcagggaga ccacaacggt ttccctctac 1321 aaataatttt gtttaacttt tactagagtg ggaccctacc tgcaggtgca gtaaggagga 1381 aaaaaaaatg aacacgatta acatcgctaa gaacgacttc tctgacatcg aactggctgc 1441 tatcccgttc aacactctgg ctgaccatta cggtgagcgt ttagctcgcg aacagttggc 1501 ccttgagcat gagtcttacg agatgggtga agcacgcttc cgcaagatgt ttgagtgata 1561 acttaaagct ggtgaggttg cggataacgc tgccgccaag cctctcatca ctaccctact 1621 ccctaagatg attgcacgca tcaacgactg gtttgaggaa gtgaaagcta agcgcggcaa 1681 gcgcccgaca gccttccagt tcctgcaaga aatcaagccg gaagccgtag cgtacatcac 1741 cattaagacc actctggctt gcctaaccag tgctgacaat acaaccgttc aggctgtagc 1801 aagcgcaatc ggtcgggcca ttgaggacga ggctcgcttc ggtcgtatcc gtgaccttaa 1861 agctaagcac ttcaagaaaa acgttgagga acaactcaac aagcgcgtag ggcacgtcta 1921 caagaaagca tttatgcaag ttgtcgaggc tgacatgctc tctaagggtc tactcggtgg 1981 cgaggcgtgg tcttcgtggc ataaggaaga ctctattcat gtaggagtac gctgcatcga 2041 gatgctcatt gagtcaaccg gaatggttag cttacaccgc caaaatgctg gcgtagtagg 2101 tcaagactct gagactatcg aactcgcacc tgaatacgct gaggctatcg caacccgtgc 2161 aggtgcgctg gctggcatct ctccgatgtt ccaaccttgc gtagttcctc ctaagccgtg 2221 gactggcatt actggtggtg gctattgggc taacggtcgt cgtcctctgg cgctggtgcg 2281 tactcacagt aagaaagcac tgatgcgcta cgaagacgtt tacatgcctg aggtgtacaa 2341 agcgattaac attgcgcaaa acaccgcatg gaaaatcaac aagaaagtcc tagcggtcgc 2401 caacgtaatc accaagtgga agcattgtcc ggtcgaggac atccctgcga ttgagcgtga 2461 agaactcccg atgaaaccgg aagacatcga catgaatcct gaggctctca ccgcgtggaa 2521 acgtgctgcc gctgctgtgt accgcaagga caaggctcgc aagtctcgcc gtatcagcct 2581 tgagttcatg cttgagcaag ccaataagtt tgctaaccat aaggccatct ggttccctta 2641 caacatggac tggcgcggtc gtgtttacgc tgtgtcaatg ttcaacccgc aaggtaacga 2701 tatgaccaaa ggactgctta cgctggcgaa aggtaaacca atcggtaagg aaggttacta 2761 ctggctgaaa atccacggtg caaactgtgc gggtgtcgat aaggttccgt tccctgagcg 2821 catcaagttc attgaggaaa accacgagaa catcatggct tgcgctaagt ctccactgga 2881 gaacacttgg tgggctgagc aagattctcc gttctgcttc cttgcgttct gctttgagta 2941 cgctggggta cagcaccacg gcctgagcta taactgctcc cttccgctgg cgtttgacgg 3001 gtcttgctct ggcatccagc acttctccgc gatgctccga gatgaggtag gtggtcgcgc 3061 ggttaacttg cttcctagtg aaaccgttca ggacatctac gggattgttg ctaagaaagt 3121 caacgagatt ctacaagcag acgcaatcaa tgggaccgat aacgaagtag ttaccgtgac 3181 cgatgagaac actggtgaaa tctctgagaa agtcaagctg ggcactaagg cactggctgg 3241 tcaatggctg gcttacggtg ttactcgcag tgtgactaag cgttcagtca tgacgctggc 3301 ttacgggtcc aaagagttcg gcttccgtca acaagtgctg gaagatacca ttcagccagc 3361 tattgattcc ggcaagggtc tgatgttcac tcagccgaat caggctgctg gatacatggc 3421 taagctgatt tgggaatctg tgagcgtgac ggtggtagct gcggttgaag caatgaactg 3481 gcttaagtct gctgctaagc tgctggctgc tgaggtcaaa gataagaaga ctggagagat 3541 tcttcgcaag cgttgcgctg tgcattgggt aactcctgat ggtttccctg tgtggcagga 3601 atacaagaag cctattcaga cgcgcttgaa cctgatgttc ctcggtcagt tccgcttaca 3661 gcctaccatt aacaccaaca aagatagcga gattgatgca cacaaacagg agtctggtat 3721 cgctcctaac tttgtacaca gccaagacgg tagccacctt cgtaagactg tagtgtgggc 3781 acacgagaag tacggtatcg aatcttttgc actgattcac gactccttcg gtaccattcc 3841 ggctgacgct gcgaacctgt tcaaagcagt gcgcgaaact atggttgaca catatgagtc 3901 ttgtgatgta ctggctgatt tctacgacca gttcgctgac cagttgcacg agtctcaatt 3961 ggacaaaatg ccagcacttc cggctaaagg taacttgaac ctccgtgaca tcttagagtc 4021 cgacttcgcg ttcgcctaat ggcactcctc agcaaatata atgaccctct tgataaccca 4081 agagggcatt ttttaatgcc catggcgttt atttgccgac taccttggtg atctcgcctt 4141 tcacgtagtg gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt 4201 gtccaagata agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct 4261 ccattgccca gtcggcagcg acatccttcg gcgcgatttt gccggttact gcgctgtacc 4321 aaatgcggga caacgtaagc actacatttc gctcatcgcc agcccagtcg ggcggcgagt 4381 tccatagcgt taaggtttca tttagcgcct caaatagatc ctgttcagga accggatcaa 4441 agagttcctc cgccgctgga cctaccaagg caacgctatg ttctcttgct tttgtcagca 4501 agatagccag atcaatgtcg atcgtggctg gctcgaagat acctgcaaga atgtcattgc 4561 gctgccattc tccaaattgc agttcgcgct tagctggata acgccacgga atgatgtcgt 4621 cgtgcacaac aatggtgact tctacagcgc ggagaatctc gctctctcca ggggaagccg 4681 aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc cttacggtca 4741 ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc gccatccact gcggagccgt 4801 acaaatgtac ggccagcaac gtcggttcga gatggcgctc gatgacgcca actacctctg 4861 atagttgagt cgatacttcg gcgatcaccg cttccctcat actcttcctt tttcaatatt 4921 attgaagcat ttatcagggt tattgtctca tgagcgtata catatttgaa tgtatttaga 4981 aaaataggcc aaatagggtt tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 5041 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccacctcttt 5101 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc 5161 cgtagttagg ccaccacttc aagaactc //