LOCUS Exported 4301 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Intended for co-expression with human Ser/Thr kinase plasmids to enhance bacterial kinase expression.. ACCESSION . VERSION . KEYWORDS Lambda Phosphatase SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4301) AUTHORS Albanese SK, Parton DL, Isik M, Rodriguez-Laureano L, Hanson SM, Behr JM, Gradia S, Jeans C, Levinson NM, Seeliger MA, Chodera JD TITLE An open library of human kinase domain constructs for automated bacterial expression. JOURNAL Biochemistry. 2018 Jul 13. doi: 10.1021/acs.biochem.7b01081. PUBMED 30004690 REFERENCE 2 (bases 1 to 4301) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4301 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 11..30 /label=T7 /note="T7 promoter, forward primer" promoter 11..29 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 34..57 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." RBS 110..132 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" primer_bind complement(931..949) /label=T7 Term /note="T7 terminator, reverse primer" terminator 945..992 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS complement(1163..1954) /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(1955..2046) /gene="bla" /label=AmpR promoter CDS complement(3106..4188) /codon_start=1 /gene="lacI" /product="lac repressor" /label=lacI /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind 4150..4169 /label=LacI-R /note="LacI, reverse primer" promoter complement(4189..4266) /gene="lacI" /label=lacI promoter ORIGIN 1 attaggaaat taatacgact cactataggg agagaattgt gagcggataa caattcccct 61 gtagattaat taagcggccg ccctgcagga ctcgagttct agaaataatt ttgtttaact 121 ttaagaagga gatatagatc atgcggtact acgaaaaaat cgacggtagc aaatatcgca 181 atatctgggt tgtgggcgac ctgcatggct gctatacaaa tctgatgaac aagcttgaca 241 ctatcgggtt cgataataaa aaagacctgc tgatcagcgt gggagatctg gttgaccgtg 301 gggctgaaaa cgttgaatgc ctggaactga tcaccttccc ctggttccgc gccgtgcgtg 361 gcaaccatga acagatgatg atcgatggac tgagcgaacg cggtaatgtg aaccattggc 421 tgttaaatgg aggaggctgg tttttcaatc tggactacga caaagagatc ctggccaaag 481 ccttggcaca taaagcagat gagctgcctc tcatcattga actggttagt aaagataaaa 541 aatatgtgat ttgccacgcg gattaccctt ttgacgagta tgaatttgga aaaccggtgg 601 atcatcagca ggttatttgg aaccgcgagc gcatctctaa ttcgcagaat ggcatcgtga 661 aggaaattaa gggcgccgat accttcatct tcggtcatac gccagcagtg aaacccctga 721 agtttgcgaa ccagatgtat attgataccg gcgcggtttt ctgcggtaat ttgacgctca 781 tccaggttca gggcgaaggt gcctaataag atcccaactc cataaggatc cgcgatcgcg 841 gcgcgccacc tggtggccgg ccggtaccac gcgtgcgcgc tgatccggct gctaacaaag 901 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg 961 gggcctctaa acgggtcttg aggggttttt tgctgaaacc tcaggcattt gagaagcaca 1021 cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta 1081 tttaacgacc ctgccctgaa ccgacgaccg ggtcatcgtg gccggatctt gcggcccctc 1141 ggcttgaacg aattgttaga cattatttgc cgactacctt ggtgatctcg cctttcacgt 1201 aatggacaaa ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa 1261 gataagcctg tctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg 1321 cccagtcggc agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc 1381 gggacaacgt aagcactaca tttcgctcat cgccagccca gtcgggcggc gagttccata 1441 gcgttaaggt ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt 1501 cctccgccgc tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag 1561 ccagatcaat gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc 1621 attctccaaa ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca 1681 caacaatggt gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt 1741 ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa 1801 ccagcaaatc aatatcactg tgcggcttca ggccgccatc cactgcggag ccgtacaaat 1861 gtacggccag caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt 1921 gagtcgatac ttcggcgatc accgcttccc tcatactctt cctttttcat tattattgaa 1981 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 2041 aacaaatagc tagctcactc ggtcgctacg ctccgggcgt gagactgcgg cgggcgctgc 2101 ggacacatac aaagttaccc acagattccg tggataagca ggggactaac atgtgaggca 2161 aaacagcagg gccgcgccgg tggcgttttt ccataggctc cgccctcctg ccagagttca 2221 cataaacaga cgcttttccg gtgcatctgt gggagccgtg aggctcaacc atgaatctga 2281 cagtacgggc gaaacccgac aggacttaaa gatccccacc gtttccggcg ggtcgctccc 2341 tcttgcgctc tcctgttccg accctgccgt ttaccggata cctgttccgc ctttctccct 2401 tacgggaagt gtggcgcttt ctcatagctc acacactggt atctcggctc ggtgtaggtc 2461 gttcgctcca agctgggctg taagcaagaa ctccccgttc agcccgactg ctgcgcctta 2521 tccggtaact gttcacttga gtccaacccg gaaaagcacg gtaaaacgcc actggcagca 2581 gccattggta actgggagtt cgcagaggat ttgtttagct aaacacgcgg ttgctcttga 2641 agtgtgcgcc aaagtccggc tacactggaa ggacagattt ggttgctgtg ctctgcgaaa 2701 gccagttacc acggttaagc agttccccaa ctgacttaac cttcgatcaa accacctccc 2761 caggtggttt tttcgaaggt taagtcagtt ggggaactgc ttaaccttcg atcaaaccac 2821 ctccccaggt ggttttttcg tttacagggc aaaagattac gcgcagaaaa aaaggatctc 2881 aagaagatcc tttgatcttt tctactgaac cgctcttatt tcagtgcaat ttatctcttc 2941 aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat gttagtcatg 3001 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat 3061 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc 3121 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 3181 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg 3241 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc 3301 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc 3361 ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat 3421 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat 3481 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc 3541 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg 3601 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa 3661 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt 3721 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc 3781 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg 3841 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga 3901 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga 3961 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc 4021 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt 4081 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata 4141 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct 4201 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat 4261 ggtgtccggg atctcgacgc tctcccttat gcgactcctg c //