LOCUS Exported 6727 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION expresses protein UbcH7 (C86K) fused to Bactrocera parkin with residues 1-29, 117-160 and 412-496 deleted. ACCESSION . VERSION . KEYWORDS pGEX-UbcH7(C86K)-10aa-linker-delta 1-29, 117-160 and 412-496 B... SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6727) AUTHORS Sauve V, Sung G, Soya N, Kozlov G, Blaimschein N, Miotto LS, Trempe JF, Lukacs GL, Gehring K TITLE Mechanism of parkin activation by phosphorylation. JOURNAL Nat Struct Mol Biol. 2018 Jul;25(7):623-630. doi: 10.1038/s41594-018-0088-7. Epub 2018 Jul 2. PUBMED 29967542 REFERENCE 2 (bases 1 to 6727) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6727 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 247..264 /label=L4440 /note="L4440 vector, forward primer" promoter 337..414 /gene="lacI" /label=lacI promoter CDS 415..1497 /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 complement(434..453) /label=LacI-R /note="LacI, reverse primer" promoter 1546..1576 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1584..1600 /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)." primer_bind 1608..1624 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 1608..1624 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 1620..2147 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR QPSEAVVWLCRS" primer_bind complement(1640..1657) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(1640..1656) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1649..1671) /label=M13/pUC Forward /note="In lacZ gene" promoter 2249..2277 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 2285..2301 /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)." CDS 2324..2977 /codon_start=1 /product="glutathione S-transferase from Schistosoma japonicum" /label=GST /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK" primer_bind 2935..2957 /label=pGEX 5' /note="pGEX vectors, Glutathione-S-transferase, forward primer" CDS 2984..3007 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" regulatory 3015..3024 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(4843..4865) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5010..5028) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5096..5200 /gene="bla" /label=AmpR promoter CDS 5201..6061 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(5419..5438) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6232..93 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6721..13 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg 61 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg 121 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct 181 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc 241 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt 301 acgcatctgt gcggtatttc acaccgcata aattccgaca ccatcgaatg gcgcaaaacc 361 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa 421 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc 481 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg 541 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg 601 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg 661 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc 721 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg 781 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat 841 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc 901 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc 961 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat 1021 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc 1081 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg 1141 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg 1201 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat 1261 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac 1321 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca 1381 ctggtgaaaa gaaaaaccac gctagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg 1441 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg 1501 caacgcaatt aatgtaagtt agctcactca ttaggcaccc caggctttac actttatgct 1561 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta 1621 tgaccatgat tacggattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg 1681 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg 1741 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgct 1801 ttgcctggtt tccggcacca gaagcggtgc cggaaagctg gctggagtgc gatcttcctg 1861 aggccgatac tgtcgtcgtc ccctcaaact ggcagatgca cggttacgat gcgcccatct 1921 acaccaacgt gacctatccc attacggtca atccgccgtt tgttcccacg gagaatccga 1981 cgggttgtta ctcgctcaca tttaatgttg atgaaagctg gctacaggaa ggccagacgc 2041 gaattatttt tgatggcgtt ggaattagct tatcgactgc acggtgcacc aatgcttctg 2101 gcgtcaggca gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat 2161 tcgtgtcgct caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg 2221 gttctggcaa atattctgaa atgagctgtt gacaattaat catcggctcg tataatgtgt 2281 ggaattgtga gcggataaca atttcacaca ggaaacagta ttcatgtccc ctatactagg 2341 ttattggaaa attaagggcc ttgtgcaacc cactcgactt cttttggaat atcttgaaga 2401 aaaatatgaa gagcatttgt atgagcgcga tgaaggtgat aaatggcgaa acaaaaagtt 2461 tgaattgggt ttggagtttc ccaatcttcc ttattatatt gatggtgatg ttaaattaac 2521 acagtctatg gccatcatac gttatatagc tgacaagcac aacatgttgg gtggttgtcc 2581 aaaagagcgt gcagagattt caatgcttga aggagcggtt ttggatatta gatacggtgt 2641 ttcgagaatt gcatatagta aagactttga aactctcaaa gttgattttc ttagcaagct 2701 acctgaaatg ctgaaaatgt tcgaagatcg tttatgtcat aaaacatatt taaatggtga 2761 tcatgtaacc catcctgact tcatgttgta tgacgctctt gatgttgttt tatacatgga 2821 cccaatgtgc ctggatgcgt tcccaaaatt agtttgtttt aaaaaacgta ttgaagctat 2881 cccacaaatt gataagtact tgaaatccag caagtatata gcatggcctt tgcagggctg 2941 gcaagccacg tttggtggtg gcgaccatcc tccaaaatcg gatctggaag ttctgttcca 3001 ggggcccctg ggatccatgg cggccagcag gaggctgatg aaggagcttg aagaaatccg 3061 caaatgtggg atgaaaaact tccgtaacat ccaggttgat gaagctaatt tattgacttg 3121 gcaagggctt attgttcctg acaaccctcc atatgataag ggagccttca gaatcgaaat 3181 caactttcca gcagagtacc cattcaaacc accgaagatc acatttaaaa caaagatcta 3241 tcacccaaac atcgacgaaa aggggcaggt caaactgcca gtaattagtg ccgaaaactg 3301 gaagccagca accaaaaccg accaagtaat ccagtccctc atagcactgg tgaatgaccc 3361 ccagcctgag cacccgcttc gggctgacct agctgaagaa tactctaagg accgtaaaaa 3421 attctgtaag aatgctgaag agtttacaaa gaaatatggg gaaaagcgac ctgtggacgg 3481 cggtgaaaac ctgtacttgg ggggatccct gagcatctac atcaagacga acacgggtcg 3541 tacgctgagc gtaaacctgg agccacaatg ggacatcaag aacgtgaagg agatcgtggc 3601 acctcagctg ggtctgcaac ctgaggaggt aaagatcatc ttcgccggca aggagctgag 3661 cgacgcaact actatccagg agtgtgacct gggtcagcaa agcatcctgc atgcaatccg 3721 ttctcgtcct caaccacagc gtcagcgttc tgaggaacgt atcaacatca ccgaggagga 3781 ccgtcagcgt actaaagcac acttcttcgt ccactgcgca cagtgcaaca aactgtgcaa 3841 aggcaaactg cgcgtgcgtt gctctctgtg taaaggcggt gctttcaccg tacaccgtga 3901 cccggaatgt tgggatgacg ttctgaaacc gcgtcgtatc actggtcatt gcgaatccca 3961 ggaaatcgcc tgcttcgaca acgaaaccgg tgacccgccg tttactgaat tctacttcaa 4021 atgcggcgaa catgtgtccg gcggtgaaaa agacttcgcg gcgccgctga acctgattaa 4081 aattaacatt aaagatgtcc cgtgcctggc ttgcacggaa gtgagcgaaa ccgtgctggt 4141 gttcccgtgc gaaagcaaac acgtcacctg cctggaatgc ttcgaacagt actgccgctc 4201 ccgcctgtcc gaacgtcagt tcatgccgca tccggatatt ggttacaccc tgccgtgtcc 4261 ggctggttgt gaaaactcct tcattgaaga aattcaccac ttcaaactgc tgtcccgcga 4321 agaatacgcg cgctaccagc gttttgcgac cgaagaatac gttctgcagg cgggcggtgt 4381 tctgtgcccg cagccgggtt gtggtatggg tctgctggtt gaaccggaat gtaaaaaagt 4441 tacctgccag aacggttgcg gttatgtttt ttgccgcaac tgcctgcagg gctatcacct 4501 gggtgattgc ctgccggaat aaacttctac caatgcttcc ggctcttgcg aatattctgt 4561 tgatccgaat cgcgcggctg aagctcgttg ggatgaagcg tctaaagtta ccattaaagt 4621 tatgaccaaa ccgtgcccga aatgccgcac cccgaccgaa cgtgatggcg gcgctatgca 4681 catggtttgt acccgcgccg gctgtggctt tgaatggtgt tggatttgtc agaccgaatg 4741 gacccgtgat tgtatgggcg cccactggtt tggctaactc gagcggccgc atcgtgactg 4801 actgacgatc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 4861 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 4921 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag 4981 cggagtgtat aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa 5041 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg 5101 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata 5161 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg 5221 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac 5281 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact 5341 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat 5401 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga 5461 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac 5521 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat 5581 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac 5641 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct 5701 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac 5761 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga 5821 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg 5881 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact 5941 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac 6001 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta 6061 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt 6121 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga 6181 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc 6241 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt 6301 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc 6361 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc 6421 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 6481 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 6541 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga 6601 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc 6661 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 6721 gggaaac //