LOCUS Exported 6096 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression of human Park2 fused to EYFP. ACCESSION . VERSION . KEYWORDS YFP-Parkin SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6096) AUTHORS Narendra D, Tanaka A, Suen DF, Youle RJ TITLE Parkin is recruited selectively to impaired mitochondria and promotes their autophagy. JOURNAL J Cell Biol. 2008 Dec 1. 183(5):795-803. PUBMED 19029340 REFERENCE 2 (bases 1 to 6096) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6096 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 607..616 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 613..1329 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(658..679) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(919..938) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1266..1287 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2884..3005 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2921..2940) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2975..2994 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3012..3467) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3149..3170) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3361..3380 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3494..3598 /gene="bla" /label=AmpR promoter promoter 3600..3957 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3623..3643) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3808..3943 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3870..3889 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3992..4786 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4046..4065) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4656..4675 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4974..4993) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5018..5065 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5394..5982 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5883..5902 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 781 ccctggccca ccctcgtgac caccttcggc tacggcctgc agtgcttcgc ccgctacccc 841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac 1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc 1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1201 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc 1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 1321 ctgtacaagt ccggactcag atctcgagct caagctttga tagtgtttgt caggttcaac 1381 tccagccatg gtttcccagt ggaggtcgat tctgacacca gcatcttcca gctcaaggag 1441 gtggttgcta agcgacaggg ggttccggct gaccagttgc gtgtgatttt cgcagggaag 1501 gagctgagga atgactggac tgtgcagaat tgtgacctgg atcagcagag cattgttcac 1561 attgtgcaga gaccgtggag aaaaggtcaa gaaatgaatg caactggagg cgacgacccc 1621 agaaacgcgg cgggaggctg tgagcgggag ccccagagct tgactcgggt ggacctcagc 1681 agctcagtcc tcccaggaga ctctgtgggg ctggctgtca ttctgcacac tgacagcagg 1741 aaggactcac caccagctgg aagtccagca ggtagatcaa tctacaacag cttttatgtg 1801 tattgcaaag gcccctgtca aagagtgcag ccgggaaaac tcagggtaca gtgcagcacc 1861 tgcaggcagg caacgctcac cttgacccag ggtccatctt gctgggatga tgttttaatt 1921 ccaaaccgga tgagtggtga atgccaatcc ccacactgcc ctgggactag tgcagaattt 1981 ttctttaaat gtggagcaca ccccacctct gacaaggaaa catcagtagc tttgcacctg 2041 atcgcaacaa atagtcggaa catcacttgc attacgtgca cagacgtcag gagccccgtc 2101 ctggttttcc agtgcaactc ccgccacgtg atttgcttag actgtttcca cttatactgt 2161 gtgacaagac tcaatgatcg gcagtttgtc cacgaccctc agcttggcta ctccctgcct 2221 tgtgtggctg gctgtcccaa ctccttgatt aaagagctcc atcacttcag gattctggga 2281 gaagagcagt acaaccggta ccagcagtat ggtgcagagg agtgtgtcct gcagatgggg 2341 ggcgtgttat gcccccgccc tggctgtgga gcggggctgc tgccggagcc tgaccagagg 2401 aaagtcacct gcgaaggggg caatggcctg ggctgtgggt ttgccttctg ccgggaatgt 2461 aaagaagcgt accatgaagg ggagtgcagt gccgtatttg aagcctcagg aacaactact 2521 caggcctaca gagtcgatga aagagccgcc gagcaggctc gttgggaagc agcctccaaa 2581 gaaaccatca agaaaaccac caagccctgt ccccgctgcc atgtaccagt ggaaaaaaat 2641 ggaggctgca tgcacatgaa gtgtccgcag ccccagtgca ggctcgagtg gtgctggaac 2701 tgtggctgcg agtggaaccg cgtctgcatg ggggaccact ggttcgacgt gtaaggatcc 2761 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct 2821 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt 2881 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 2941 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 3001 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta 3061 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga 3121 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa 3181 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga 3241 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc 3301 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga 3361 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg 3421 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt 3481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 3541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc 3601 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 3661 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 3721 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 3781 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 3841 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc 3901 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat 3961 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg 4021 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg 4081 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa 4141 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct 4201 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga 4261 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc 4321 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac 4381 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc 4441 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact 4501 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga 4561 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg 4621 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga 4681 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga 4741 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg 4801 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc 4861 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc 4921 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca 4981 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa 5041 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct 5101 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca 5161 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc 5221 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa 5281 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt 5341 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat 5401 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 5461 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga 5521 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac 5581 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt 5641 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag 5701 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc 5761 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag 5821 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca 5881 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt 5941 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc 6001 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc 6061 cctgattctg tggataaccg tattaccgcc atgcat //