LOCUS Exported 5566 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Baculovirus expression for structure determination. ACCESSION . VERSION . KEYWORDS 6OGJ SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5566) TITLE SGC Toronto 2019-Q1 to 2020-Q2 REFERENCE 2 (bases 1 to 5566) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5566 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 320..337 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 387..404 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(457..475) /label=T7 Term /note="T7 terminator, reverse primer" terminator 471..518 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 555..1010 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(642..661) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 852..873 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(1102..1917) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 1825..1844 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 1909..1928 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 2039..2627 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2528..2547 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2781..2798 /label=L4440 /note="L4440 vector, forward primer" misc_feature 2813..2952 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(2878..2897) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3038..3060 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3054..3245) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4054..5136) /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 5098..5117 /label=LacI-R /note="LacI, reverse primer" promoter complement(5137..5214) /gene="lacI" /label=lacI promoter primer_bind 5420..5439 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 5523..5542 /label=T7 /note="T7 promoter, forward primer" promoter 5523..5541 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 5542..5566 /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)." ORIGIN 1 cctctagaaa taattttgtt taaccttaag aaggagatat actatggctt ctgcctcccc 61 caaacagcgg cgctccatca tccgtgaccg gggacccatg tatgatgacc ccaccctgcc 121 tgaaggctgg acacggaagc ttaagcaaag gaaatctggc cgctctgctg ggaagtatga 181 tgtgtatttg atcaatcccc agggaaaagc ctttcgctct aaagtggagt tgattgcgta 241 cttcgaaaag gtaggcgaca catccctgga ccctaatgat tttgacttca cggtaactgg 301 gagagggagc ccctccgcgc accatcatca ccaccattga ggatccgaat tcgagctccg 361 tcgacaagct tgcggccgca ctcgagcacc accaccacca ccactgagat ccggctgcta 421 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac 481 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg 541 gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac 601 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc 661 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt 721 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg 781 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 841 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta 901 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat 961 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact 1021 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg 1081 tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt 1141 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga 1201 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac 1261 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga 1321 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt 1381 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa 1441 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg 1501 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat 1561 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc 1621 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg 1681 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct 1741 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat 1801 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc 1861 catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac 1921 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt 1981 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt 2041 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag 2101 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca 2161 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca 2221 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 2281 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 2341 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 2401 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 2461 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 2521 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 2581 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 2641 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt 2701 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc 2761 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc 2821 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac 2881 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg 2941 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg 3001 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg 3061 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg 3121 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga 3181 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg 3241 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa 3301 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt 3361 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt 3421 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct 3481 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac 3541 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag 3601 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc 3661 cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc 3721 atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag 3781 gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg 3841 ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg 3901 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac 3961 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa 4021 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 4081 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 4141 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac 4201 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa 4261 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta 4321 tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc 4381 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag 4441 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat 4501 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga 4561 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg 4621 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg 4681 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc 4741 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt 4801 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct 4861 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag 4921 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc 4981 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt 5041 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc 5101 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc 5161 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat 5221 ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc 5281 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc 5341 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc 5401 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg 5461 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa 5521 attaatacga ctcactatag gggaattgtg agcggataac aattcc //