LOCUS Exported 4741 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION photo-activatable mitochondrial marker for expression in mammalian cells. ACCESSION . VERSION . KEYWORDS mito-dendra2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4741) AUTHORS Pham AH, McCaffery JM, Chan DC TITLE Mouse lines with photo-activatable mitochondria to study mitochondrial dynamics. JOURNAL Genesis. 2012 Nov;50(11):833-43. doi: 10.1002/dvg.22050. Epub 2012 Aug 11. PUBMED 22821887 REFERENCE 2 (bases 1 to 4741) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4741 /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" CDS 597..671 /codon_start=1 /product="mitochondrial presequence of human cytochrome c oxidase subunit VIII (Rizutto et al., 1989)" /label=COX8 presequence /translation="MSVLTPLLLRGLTGSARRLPVPRAK" CDS 693..1379 /codon_start=1 /product="monomeric photoswitchable green-to-red fluorescent protein from Dendronephthya" /label=Dendra2 /note="mammalian codon-optimized" /translation="NTPGINLIKEDMRVKVHMEGNVNGHAFVIEGEGKGKPYEGTQTAN LTVKEGAPLPFSYDILTTAVHYGNRVFTKYPEDIPDYFKQSFPEGYSWERTMTFEDKGI CTIRSDISLEGDCFFQNVRFKGTNFPPNGPVMQKKTLKWEPSTEKLHVRDGLLVGNINM ALLLEGGGHYLCDFKTTYKAKKVVQLPDAHFVDHRIEILGNDSDYNKVKLYEHAVARYS PLPSQVW" polyA_signal 1529..1650 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1566..1585) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1620..1639 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1657..2112) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1794..1815) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2006..2025 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2139..2243 /gene="bla" /label=AmpR promoter promoter 2245..2602 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2268..2288) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2453..2588 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2515..2534 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2637..3431 /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(2691..2710) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3301..3320 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3619..3638) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3663..3710 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4039..4627 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4528..4547 /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 gctagcatgt 601 ccgtcctgac gccgctgctg ctgcggggct tgacaggctc ggcccggcgg ctcccagtgc 661 cgcgcgccaa gatccattcg ttgggggatc cgaacacccc gggaattaac ctgatcaagg 721 aggacatgcg cgtgaaggtg cacatggagg gcaacgtgaa cggccacgcc ttcgtgatcg 781 agggcgaggg caagggcaag ccctacgagg gcacccagac cgccaacctg accgtgaagg 841 agggcgcccc cctgcccttc agctacgaca tcctgaccac cgccgtgcac tacggcaacc 901 gggtgttcac caagtacccc gaggacatcc ccgactactt caagcagagc ttccccgagg 961 gctacagctg ggagcgcacc atgaccttcg aggacaaggg catctgcacc atccgcagcg 1021 acatcagcct ggagggcgac tgcttcttcc agaacgtgcg cttcaagggc accaacttcc 1081 cccccaacgg ccccgtgatg cagaagaaga ccctgaagtg ggagcccagc accgagaagc 1141 tgcacgtgcg cgacggcctg ctggtgggca acatcaacat ggccctgctg ctggagggcg 1201 gcggccacta cctgtgcgac ttcaagacca cctacaaggc caagaaggtg gtgcagctgc 1261 ccgacgccca cttcgtggac caccgcatcg agatcctggg caacgacagc gactacaaca 1321 aggtgaagct gtacgagcac gccgtggccc gctacagccc cctgcccagc caggtgtggt 1381 ccggcgacag cggcgtgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac 1441 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa 1501 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa 1561 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 1621 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt 1681 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg 1741 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt 1801 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct 1861 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt 1921 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa 1981 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc 2041 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc 2101 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat 2161 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc 2221 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca 2281 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 2341 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 2401 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc 2461 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 2521 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt 2581 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga 2641 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct 2701 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc 2761 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag 2821 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg 2881 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc 2941 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc 3001 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg 3061 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc 3121 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg 3181 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc 3241 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag 3301 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg 3361 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg 3421 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc 3481 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt 3541 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca 3601 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat 3661 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg 3721 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata 3781 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct 3841 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta 3901 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa 3961 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga 4021 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac 4081 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt 4141 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc 4201 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat 4261 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag 4321 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc 4381 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag 4441 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac 4501 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg 4561 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct 4621 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc 4681 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca 4741 t //