LOCUS Exported 6898 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Bacterial expression for structure determination; may not be full ORF. ACCESSION . VERSION . KEYWORDS 5JJW (USP15_SART3) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6898) TITLE SGC Toronto Q2 Q3 2016 REFERENCE 2 (bases 1 to 6898) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6898 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..20 /label=T7 /note="T7 promoter, forward primer" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 20..44 /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 59..81 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 91..108 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 121..141 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 1763..1780 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1833..1851) /label=T7 Term /note="T7 terminator, reverse primer" terminator 1847..1894 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 1931..2386 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2018..2037) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2228..2249 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(2478..3293) /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 3201..3220 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 3285..3304 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 3415..4003 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3904..3923 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4157..4174 /label=L4440 /note="L4440 vector, forward primer" misc_feature 4189..4328 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(4254..4273) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4414..4436 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(4430..4621) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(5430..6512) /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 6474..6493 /label=LacI-R /note="LacI, reverse primer" promoter complement(6513..6590) /gene="lacI" /label=lacI promoter primer_bind 6796..6815 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt 61 tgtttaactt taagaaggag atataccatg catcatcatc atcatcacag cagcggcaga 121 gaaaacttgt atttccaggg catggcggaa ggcggagcgg cggatctgga cacccagcgg 181 tctgacatcg cgacgctgct caaaacctcg ctccggaaag gggacacctg gtacctagtc 241 gatagtcgct ggttcaaaca gtggaaaaaa tatgttggct ttgacagttg ggacaaatac 301 cagatgggag atcaaaatgt gtatcctgga cccattgata actctggact tctcaaagat 361 ggtgatgccc agtcacttaa ggaacacctt attgatgaat tggattacat actgttgcca 421 actgaaggtt ggaataaact tgtcagctgg tacacattga tggaaggtca agagccaata 481 gcacgaaagg tggttgaaca gggtatgttt gtaaagcact gcaaagtaga agtatatctc 541 acagaattga agctatgtga aaatggaaac atgaataatg ttgtaactcg aagatttagc 601 aaagctgaca caatagatac aattgaaaag gaaataagaa aaatcttcag tattccagat 661 gaaaaggaga ccagattgtg gaacaaatac atgagtaaca catttgaacc actgaataaa 721 ccagacagca ccattcagga tgctggttta taccaaggac aggtattagt gatagaacag 781 aaaaatgaag atggaacatg gccaaggggt tgactcgacg gaggaataat catatgtcag 841 taattcagaa ctataacaaa gcactacagc agctggagaa atataaaccc tatgaagaag 901 cactgttgca ggcagaggca ccaaggctgg cagaatatca agcatatatc gattttgaga 961 tgaaaattgg cgatcctgct cgcattcagt tgatctttga gcgcgccctg gtcgagaact 1021 gccttgtccc agacttatgg atccgttaca gtcagtacct agatcgacaa ctgaaagtaa 1081 aggatttggt tttatctgta cataaccgcg ctattagaaa ctgcccctgg acagttgcct 1141 tatggagtcg gtacctcttg gccatggaga gacatggagt tgatcatcaa gtaatttctg 1201 taaccttcga gaaagctttg aatgccggct tcatccaggc cactgattat gtggagattt 1261 ggcaggcata ccttgattac ctgaggagaa gggttgattt caaacaagac tccagtaaag 1321 agctggagga gttgagggcc gcctttactc gtgccttgga gtatctgaag caggaggtgg 1381 aagagcgttt caatgagagt ggtgatccaa gctgcgtgat tatgcagaac tgggctagga 1441 ttgaggctcg actgtgcaat aacatgcaga aagctcggga actctgggat agcatcatga 1501 ccagaggaaa tgccaagtac gccaacatgt ggctagagta ttacaacctg gaaagagctc 1561 atggtgacac ccagcactgc cggaaggctc tgcaccgggc cgtccagtgc accagtgact 1621 acccagagca cgtctgcgaa gtgttactca ccatggagag gacagaaggt tctttagaag 1681 attgggatat agctgttcag aaaactgaaa cccgattagc tcgtgtcaat gagtgatgac 1741 gaagcttgcg gccgcactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa 1801 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct 1861 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt 1921 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 1981 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 2041 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg 2101 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 2161 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 2221 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct 2281 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa 2341 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc 2401 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc 2461 cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc 2521 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac 2581 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt 2641 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa 2701 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag 2761 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg 2821 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa 2881 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt 2941 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg 3001 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata 3061 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct 3121 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc 3181 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg 3241 ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct cataacaccc 3301 cttgtattac tgtttatgta agcagacagt tttattgttc atgaccaaaa tcccttaacg 3361 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga 3421 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt 3481 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag 3541 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa 3601 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag 3661 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca 3721 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac 3781 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa 3841 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc 3901 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg 3961 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc 4021 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc 4081 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag 4141 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta 4201 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc 4261 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca 4321 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc 4381 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt 4441 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa 4501 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg 4561 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca 4621 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag 4681 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg 4741 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat 4801 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga 4861 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa 4921 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt 4981 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc 5041 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc 5101 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt 5161 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc 5221 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt 5281 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga 5341 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag 5401 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 5461 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 5521 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc 5581 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc 5641 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc 5701 actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc 5761 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt 5821 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc 5881 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca 5941 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc 6001 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca 6061 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc 6121 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc 6181 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca 6241 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc 6301 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg 6361 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca 6421 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac 6481 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc 6541 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg 6601 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt 6661 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc 6721 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc 6781 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc 6841 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaat //