LOCUS Exported 5434 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Encodes for a fusion protein between a mitochondrial anchor and the CIBN-GFP protein.. ACCESSION . VERSION . KEYWORDS mito-CIBN-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5434) AUTHORS Valon L, Marin-Llaurado A, Wyatt T, Charras G, Trepat X TITLE Optogenetic control of cellular forces and mechanotransduction. JOURNAL Nat Commun. 2017 Feb 10;8:14396. doi: 10.1038/ncomms14396. PUBMED 28186127 REFERENCE 2 (bases 1 to 5434) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5434 /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 1329..1338 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1335..1853 /codon_start=1 /product="N-terminal fragment of mVenus for use in bimolecular fluorescence complementation (BiFC) (Kodama and Hu, 2010)" /label=VN173 /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIE" primer_bind complement(1380..1401) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1641..1660) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1988..2009 /label=EGFP-C /note="EGFP, forward primer" misc_feature 2046..2098 /label=MCS /note="multiple cloning site" polyA_signal 2222..2343 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2259..2278) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2313..2332 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2350..2805) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2487..2508) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2699..2718 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2832..2936 /gene="bla" /label=AmpR promoter promoter 2938..3295 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2961..2981) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3146..3281 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3208..3227 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3330..4124 /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(3384..3403) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3994..4013 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4312..4331) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4356..4403 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4732..5320 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5221..5240 /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 gctagcatga 601 gtctgacttc cagttccagc gtacgagttg aatggatcgc agcagttacc attgctgctg 661 ggacagctgc aattggttat ctagcttaca aaagatttta tgttaaagat catcgaaata 721 aagctatgat aaaccttcac atccagaaag acaaccccaa gatagtacat gctggaagct 781 taccggtcgc cgaattcggt tctagaatga atggagctat aggaggtgac cttttgctca 841 attttcctga catgtcggtc ctagagcgcc aaagggctca cctcaagtac ctcaatccca 901 cctttgattc tcctctcgcc ggcttctttg ccgattcttc aatgattacc ggcggcgaga 961 tggacagcta tctttcgact gccggtttga atcttccgat gatgtacggt gagacgacgg 1021 tggaaggtga ttcaagactc tcaatttcgc cggaaacgac gcttgggact ggaaatttca 1081 aggcagcgaa gtttgataca gagactaagg attgtaatga ggcggcgaag aagatgacga 1141 tgaacagaga tgacctagta gaagaaggag aagaagagaa gtcgaaaata acagagcaaa 1201 acaatgggag cacaaaaagc atcaagaaga tgaaacacaa agccaagaaa gaagagaaca 1261 atttctctaa tgattcatct aaagtgacga aggaattgga gaaaacggat tatattcatg 1321 taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc 1381 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg 1441 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg 1501 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc 1561 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg 1621 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg 1681 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca 1741 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg 1801 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca 1861 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc 1921 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc 1981 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg 2041 agtagctcga gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac 2101 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt 2161 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt 2221 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 2281 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 2341 ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa 2401 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat 2461 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg 2521 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac 2581 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta 2641 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag 2701 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg 2761 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc 2821 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc 2881 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg 2941 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc 3001 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa 3061 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 3121 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc 3181 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 3241 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg 3301 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt 3361 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct 3421 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga 3481 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg 3541 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact 3601 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg 3661 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct 3721 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg 3781 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt 3841 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg 3901 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc 3961 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag 4021 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt 4081 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt 4141 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc 4201 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca 4261 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg 4321 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac 4381 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt 4441 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc 4501 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc 4561 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt 4621 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga 4681 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc 4741 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt 4801 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc 4861 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc 4921 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 4981 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 5041 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga 5101 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc 5161 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 5221 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg 5281 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt 5341 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc 5401 tgattctgtg gataaccgta ttaccgccat gcat //