LOCUS Exported 5489 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Baculovirus expression for structure determination. ACCESSION . VERSION . KEYWORDS 6V41 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5489) TITLE SGC Toronto 2019-Q1 to 2020-Q2 REFERENCE 2 (bases 1 to 5489) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5489 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 13..33 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 246..263 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(316..334) /label=T7 Term /note="T7 terminator, reverse primer" terminator 330..377 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 414..869 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(501..520) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 711..732 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(961..1776) /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 1684..1703 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 1768..1787 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 1898..2486 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2387..2406 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2640..2657 /label=L4440 /note="L4440 vector, forward primer" misc_feature 2672..2811 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(2737..2756) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2897..2919 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(2913..3104) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(3913..4995) /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 4957..4976 /label=LacI-R /note="LacI, reverse primer" promoter complement(4996..5073) /gene="lacI" /label=lacI promoter primer_bind 5279..5298 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 5382..5401 /label=T7 /note="T7 promoter, forward primer" promoter 5382..5400 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 5401..5425 /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 5440..5462 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 5472..5489 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" ORIGIN 1 agcagcggca gagaaaactt gtatttccag ggcgcttccc aggagtttga ggttgaagct 61 attgttgaca aaagacagga taaaaatggg aatacacagt atttggttcg gtggaaaggt 121 tatgacaaac aggatgacac ttgggaacca gagcagcacc tcatgaactg tgaaaaatgt 181 gtacatgatt ttaatagacg acagactgaa cagaaatgat gacgaagctt gcggccgcac 241 tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg 301 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg 361 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc 421 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 481 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 541 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 601 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 661 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 721 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 781 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 841 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga 901 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc 961 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat 1021 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca 1081 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc 1141 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac 1201 tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca 1261 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg 1321 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga 1381 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata 1441 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc 1501 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt 1561 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa 1621 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac 1681 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg 1741 cctagagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat 1801 gtaagcagac agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact 1861 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 1921 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 1981 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 2041 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 2101 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 2161 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 2221 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 2281 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 2341 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 2401 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 2461 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 2521 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata 2581 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca 2641 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc 2701 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata 2761 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac 2821 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga 2881 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa 2941 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct 3001 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg 3061 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa 3121 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata 3181 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg 3241 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat 3301 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata 3361 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc 3421 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg 3481 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc 3541 aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc 3601 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag 3661 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg 3721 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca 3781 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg 3841 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta 3901 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 3961 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct 4021 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg 4081 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa 4141 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc 4201 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt 4261 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa 4321 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt 4381 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc 4441 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc 4501 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa 4561 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata 4621 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc 4681 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg 4741 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac 4801 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt 4861 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg 4921 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa 4981 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat 5041 accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg 5101 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa 5161 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca 5221 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg 5281 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga 5341 tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa ttaatacgac tcactatagg 5401 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga 5461 gatataccat gcatcatcat catcatcac //