LOCUS Exported 4674 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Visualization of peroxisomes. ACCESSION . VERSION . KEYWORDS mNeonGreen-peroxi SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4674) AUTHORS Chertkova AO, Mastop M, Postma M, van Bommel N, van der Niet S, Batenburg KL, Joosen L, Gadella TWJ, Okada Y, Goedhart J TITLE Robust and Bright Genetically Encoded Fluorescent Markers for Highlighting Structures and Compartments in Mammalian Cells JOURNAL bioRxiv 160374 REFERENCE 2 (bases 1 to 4674) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4674 /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..1320 /codon_start=1 /product="bright monomeric yellow-green fluorescent protein derived from LanYFP (Shaner et al., 2013)" /label=mNeonGreen /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ KAFTDVMGMDELYK" regulatory 868..877 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" polyA_signal 1462..1583 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1499..1518) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1553..1572 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1590..2045) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1727..1748) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1939..1958 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2072..2176 /gene="bla" /label=AmpR promoter promoter 2178..2535 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2201..2221) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2386..2521 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2448..2467 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2570..3364 /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(2624..2643) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3234..3253 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3552..3571) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3596..3643 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 3972..4560 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4461..4480 /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 gaggataaca tggcctctct cccagcgaca 661 catgagttac acatctttgg ctccatcaac ggtgtggact ttgacatggt gggtcagggc 721 accggcaatc caaatgatgg ttatgaggag ttaaacctga agtccaccaa gggtgacctc 781 cagttctccc cctggattct ggtccctcat atcgggtatg gcttccatca gtacctgccc 841 taccctgacg ggatgtcgcc tttccaggcc gccatggtag atggctccgg ctaccaagtc 901 catcgcacaa tgcagtttga agatggtgcc tcccttactg ttaactaccg ctacacctac 961 gagggaagcc acatcaaagg agaggcccag gtgaagggga ctggtttccc tgctgacggt 1021 cctgtgatga ccaactcgct gaccgctgcg gactggtgca ggtcgaagaa gacttacccc 1081 aacgacaaaa ccatcatcag tacctttaag tggagttaca ccactggaaa tggcaagcgc 1141 taccggagca ctgcgcggac cacctacacc tttgccaagc caatggcggc taactatctg 1201 aagaaccagc cgatgtacgt gttccgtaag acggagctca agcactccaa gaccgagctc 1261 aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg gcatggacga gctgtacaag 1321 tcccgcctct aaggatccac cggatctaga taactgatca taatcagcca taccacattt 1381 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa 1441 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc 1501 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 1561 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt 1621 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat 1681 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa 1741 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg 1801 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa 1861 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc 1921 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag 1981 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg 2041 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 2101 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 2161 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg 2221 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 2281 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 2341 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 2401 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 2461 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 2521 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca 2581 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg 2641 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg 2701 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc 2761 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt 2821 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc 2881 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca 2941 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc 3001 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg 3061 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct 3121 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc 3181 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc 3241 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta 3301 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt 3361 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga 3421 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac 3481 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg 3541 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca 3601 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg 3661 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc 3721 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc 3781 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat 3841 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 3901 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 3961 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 4021 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 4081 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta 4141 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 4201 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 4261 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 4321 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 4381 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 4441 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 4501 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 4561 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 4621 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat //