LOCUS Exported 6283 bp ds-DNA circular SYN 05-JAN-2022 DEFINITION Bacterial Expression of MYC3. ACCESSION . VERSION . KEYWORDS pSUMO His6 SUMO MYC3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6283) AUTHORS Zhang F, Yao J, Ke J, Zhang L, Lam VQ, Xin XF, Zhou XE, Chen J, Brunzelle J, Griffin PR, Zhou M, Xu HE, Melcher K, He SY TITLE Structural basis of JAZ repression of MYC transcription factors in jasmonate signalling. JOURNAL Nature. 2015 Sep 10;525(7568):269-73. doi: 10.1038/nature14661. Epub 2015 Aug 10. PUBMED 26258305 REFERENCE 2 (bases 1 to 6283) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 5, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6283 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 16..40 /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 55..77 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 90..107 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 111..401 /codon_start=1 /gene="S. cerevisiae SMT3 (truncated)" /product="cleavable ubiquitin-like protein tag" /label=SUMO /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR RLMEAFAKRQGKEMDSLRFLYDGIRIQADQAPEDLDMEDNDIIEAHREQIGG" CDS 1011..1028 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1081..1099) /label=T7 Term /note="T7 terminator, reverse primer" terminator 1095..1142 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 1179..1634 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1266..1285) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1476..1497 /label=F1ori-F /note="F1 origin, forward primer" promoter 1660..1764 /gene="bla" /label=AmpR promoter CDS 1765..2625 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1983..2002) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2796..3384 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3285..3304 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3538..3555 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3570..3709 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3635..3654) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3795..3817 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3811..4002) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4811..5893) /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 5855..5874 /label=LacI-R /note="LacI, reverse primer" promoter complement(5894..5971) /gene="lacI" /label=lacI promoter primer_bind 6177..6196 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 6280..16 /label=T7 /note="T7 promoter, forward primer" promoter 6280..15 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" ORIGIN 1 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt 61 taactttaag aaggagatat accatgggtc atcaccatca tcatcacggg tcggactcag 121 aagtcaatca agaagctaag ccagaggtca agccagaagt caagcctgag actcacatca 181 atttaaaggt gtccgatgga tcttcagaga tcttcttcaa gatcaaaaag accactcctt 241 taagaaggct gatggaagcg ttcgctaaaa gacagggtaa ggaaatggac tccttaagat 301 tcttgtacga cggtattaga attcaagctg atcaggcccc tgaagatttg gacatggagg 361 ataacgatat tattgaggct caccgcgaac agattggagg tggatcccag cctcagttca 421 acgaagatac tcttcaacaa cgtctccaag ctttaataga atccgccgga gaaaactgga 481 cttacgctat cttctggcag atctcacacg acttcgattc atccaccgga gataacacag 541 tgatcctcgg ctggggagat ggttactaca aaggagagga agataaagag aagaagaaga 601 acaacaccaa cacggcggag caagagcatc ggaaaagagt aatacgtgag cttaactcgt 661 taatctccgg cggaattggg gtttccgatg aatcaaacga tgaagaagta acagatactg 721 aatggttctt cttagtttcg atgactcaaa gcttcgttaa cggtgttggt ctccccggag 781 aatctttctt aaactctcgt gtgatttggt tatccgggtc tggtgcttta accgggtcgg 841 gttgtgaaag agcgggtcaa ggtcagattt acgggttaaa gacgatggtg tgtatcgcga 901 ctcaaaacgg cgtcgttgag cttggttcgt cggaggttat aagtcaaagc tcagatctga 961 tgcataaagt taacaacttg tttaatttca actaagcggc cgcactcgag caccaccacc 1021 accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca 1081 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt 1141 tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt 1201 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc 1261 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca 1321 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc 1381 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt 1441 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac 1501 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc 1561 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt 1621 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta 1681 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt 1741 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc 1801 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa 1861 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt 1921 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt 1981 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc 2041 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg 2101 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg 2161 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac 2221 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca 2281 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta 2341 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat 2401 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa 2461 tctggagccg gtgagcgtgg ttctcgcggt atcattgcag cactggggcc agatggtaag 2521 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat 2581 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt 2641 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg 2701 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga 2761 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta 2821 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 2881 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact 2941 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca 3001 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt 3061 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg 3121 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag 3181 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta 3241 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat 3301 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg 3361 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc 3421 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac 3481 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc 3541 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg 3601 tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt 3661 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc 3721 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca 3781 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg 3841 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc 3901 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat 3961 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg 4021 gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg 4081 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt 4141 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac 4201 agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat 4261 ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat 4321 tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg 4381 tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa 4441 cgacaggagc acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt 4501 ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat 4561 tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc 4621 gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt 4681 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa 4741 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat 4801 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 4861 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt 4921 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca 4981 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg 5041 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac 5101 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg 5161 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac 5221 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga 5281 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta 5341 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt 5401 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg 5461 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt 5521 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt 5581 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag 5641 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc 5701 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca 5761 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt 5821 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg 5881 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac 5941 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac 6001 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg 6061 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata 6121 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg 6181 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg 6241 cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aat //