LOCUS Exported 5073 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION In vivo visualization of the Golgi apparatus (can be used for colocalization studies). ACCESSION . VERSION . KEYWORDS mNeonGreen-Giantin SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5073) 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 5073) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5073 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 213..233 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter complement(258..362) /gene="bla" /label=AmpR promoter rep_origin 389..844 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(476..495) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 686..707 /label=F1ori-F /note="F1 origin, forward primer" polyA_signal 851..972 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(862..881) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 916..935 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS complement(1513..2220) /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 1956..1965 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 2217..2226 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter complement(2265..2468) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(2294..2314) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 2469..2772 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" rep_origin complement(2947..3535) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3027..3046) /label=pBR322ori-F /note="pBR322 origin, forward primer" polyA_signal 3864..3911 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" primer_bind 3936..3955 /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" CDS complement(4143..4937) /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(4254..4273) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 4864..4883 /label=Neo-R /note="Neomycin resistance gene, reverse primer" promoter complement(4972..256) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4986..48 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(5040..5059) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" ORIGIN 1 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag 61 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg 121 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact 181 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagctg 241 gttctttccg cctcaggact cttccttttt caatattatt gaagcattta tcagggttat 301 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 361 cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg 421 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct 481 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 541 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 601 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg 661 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac 721 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta 781 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca 841 atttacgcct taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa 901 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc 961 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg 1021 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcag 1081 ttatctagat ccggtggatc cctatagatg gcccgtaaaa cacagaatga gcaggacatg 1141 aatcattaga aagtagatgg ctgctagaag tggcactcgg gtccgtgaat gacagagtga 1201 acgcaggact cgcttccatc caacgccact ccgggtcctt cgacaactgt tgcttgtaag 1261 atctattaac agtgcctgct cctgagtgcc acaggagcca atgataggag tccgggaaga 1321 gtcccattca ctgtgctcta accgtctgat ctgctcctcg gccacagaga gagcattctc 1381 agcagccact ctttggtctc gttcttcttc cagcagcttc cttaattcat tcacttcctg 1441 tctggtgttg catagctgct gctgagcttc agaaaagctt tgctgcggtt ctcctcgaga 1501 tctgagtccg gacttgtaca gctcgtccat gcccatcaca tcggtaaagg ccttttgcca 1561 ctccttgaag ttgagctcgg tcttggagtg cttgagctcc gtcttacgga acacgtacat 1621 cggctggttc ttcagatagt tagccgccat tggcttggca aaggtgtagg tggtccgcgc 1681 agtgctccgg tagcgcttgc catttccagt ggtgtaactc cacttaaagg tactgatgat 1741 ggttttgtcg ttggggtaag tcttcttcga cctgcaccag tccgcagcgg tcagcgagtt 1801 ggtcatcaca ggaccgtcag cagggaaacc agtccccttc acctgggcct ctcctttgat 1861 gtggcttccc tcgtaggtgt agcggtagtt aacagtaagg gaggcaccat cttcaaactg 1921 cattgtgcga tggacttggt agccggagcc atctaccatg gcggcctgga aaggcgacat 1981 cccgtcaggg tagggcaggt actgatggaa gccatacccg atatgaggga ccagaatcca 2041 gggggagaac tggaggtcac ccttggtgga cttcaggttt aactcctcat aaccatcatt 2101 tggattgccg gtgccctgac ccaccatgtc aaagtccaca ccgttgatgg agccaaagat 2161 gtgtaactca tgtgtcgctg ggagagaggc catgttatcc tcctcgccct tgctcaccat 2221 ggtggcgacc ggtagcgcta gcggatctga cggttcacta aaccagctct gcttatatag 2281 acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat 2341 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg 2401 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac 2461 cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc 2521 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata 2581 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa 2641 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc 2701 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta 2761 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac 2821 tattaataac taatgcatgg cggtaatacg gttatccaca gaatcagggg ataacgcagg 2881 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 2941 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 3001 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 3061 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 3121 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 3181 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 3241 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 3301 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 3361 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc 3421 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 3481 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 3541 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 3601 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 3661 ttttaaatca atctaaagta tatatgagta acctgaggct atggcagggc ctgccgcccc 3721 gacgttggct gcgagccctg ggccttcacc cgaacttggg gggtggggtg gggaaaagga 3781 agaaacgcgg gcgtattggc cccaatgggg tctcggtggg gtatcgacag agtgccagcc 3841 ctgggaccga accccgcgtt tatgaacaaa cgacccaaca ccgtgcgttt tattctgtct 3901 ttttattgcc gtcatagcgc gggttccttc cggtattgtc tccttccgtg tttcagttag 3961 cctcccccta gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca 4021 gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc 4081 gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc 4141 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg 4201 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca 4261 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg 4321 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc 4381 acgacgagat cctcgccgtc gggcatgctc gccttgagcc tggcgaacag ttcggctggc 4441 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga 4501 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca 4561 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg 4621 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct 4681 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc 4741 cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga 4801 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt 4861 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat 4921 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcgat ctttgcaaaa 4981 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc 5041 ctcggcctct gcataaataa aaaaaattag tca //