LOCUS Exported 5360 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION In vivo visualization of Rab7 (can be used for colocalization studies). ACCESSION . VERSION . KEYWORDS mTurquoise2-Rab7(WT) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5360) 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 5360) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5360 /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 monomeric variant of CFP (Goedhart et al., 2012)" /label=mTurquoise2 /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL 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 2148..2269 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2185..2204) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2239..2258 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2276..2731) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2413..2434) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2625..2644 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2758..2862 /gene="bla" /label=AmpR promoter promoter 2864..3221 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2887..2907) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3072..3207 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3134..3153 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3256..4050 /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(3310..3329) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3920..3939 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4238..4257) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4282..4329 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4658..5246 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5147..5166 /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 caccctgtcc tggggcgtgc 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 tttagcgaca acgtctatat caccgccgac 1081 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc 1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1201 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc 1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 1321 ctgtacaagt ccggactcag atctcgagct caagcttata cctctaggaa gaaagtgttg 1381 ctgaaggtta tcatcctggg agattctgga gttggtaaga catcactcat gaaccagtat 1441 gtgaacaaga aattcagtaa tcagtacaaa gctacaatag gagcagactt tctgacaaag 1501 gaggtgatgg tggatgacag actagttaca atgcagatct gggacacagc aggccaggaa 1561 cggttccagt cccttggtgt ggccttctac agaggtgcag actgctgcgt tctggtattt 1621 gacgttactg cccccaacac attcaaaacc ctcgatagct ggagagatga gtttctcatc 1681 caggccagtc cccgggatcc tgaaaacttc cctttcgttg tgttgggaaa caagattgac 1741 ctcgaaaaca gacaagtggc cacaaagcgg gcacaggcct ggtgctacag caaaaacaac 1801 attccctact tcgagaccag tgccaaggag gccatcaatg tggagcaggc gttccagacg 1861 attgcaagga atgcacttaa acaggaaaca gaggtggagc tgtacaatga attccctgaa 1921 cccatcaaac tggacaagaa cgaccgggcc aagacctcag cggaaagctg cagttgctga 1981 aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccgga tctagataac 2041 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac 2101 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg 2161 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 2221 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt 2281 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt 2341 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg 2401 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc 2461 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca 2521 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga 2581 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa 2641 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc 2701 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc 2761 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa 2821 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc 2881 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa 2941 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc 3001 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc 3061 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct 3121 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga 3181 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga 3241 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg 3301 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc 3361 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt 3421 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt 3481 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc 3541 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc 3601 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac 3661 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag 3721 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag 3781 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat 3841 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg 3901 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa 3961 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc 4021 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc 4081 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt 4141 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca 4201 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac 4261 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt 4321 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga 4381 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt 4441 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag 4501 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg 4561 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact 4621 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 4681 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 4741 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 4801 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 4861 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 4921 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 4981 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 5041 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 5101 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 5161 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 5221 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 5281 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata 5341 accgtattac cgccatgcat //