LOCUS Exported 5339 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION In vivo visualization of Rab7 (can be used for colocalization studies). ACCESSION . VERSION . KEYWORDS mScarletI-Rab7 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5339) 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 5339) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5339 /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..1308 /codon_start=1 /product="bright monomeric red fluorescent protein evolved from a synthetic template (Bindels et al., 2016)" /label=mScarlet-I /note="matures faster than mScarlet" /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH STGGMDELYK" primer_bind complement(746..764) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1014..1033 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 2127..2248 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2164..2183) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2218..2237 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2255..2710) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2392..2413) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2604..2623 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2737..2841 /gene="bla" /label=AmpR promoter promoter 2843..3200 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2866..2886) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3051..3186 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3113..3132 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3235..4029 /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(3289..3308) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3899..3918 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4217..4236) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4261..4308 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4637..5225 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5126..5145 /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 gcagtgatca aggagttcat gcggttcaag 661 gtgcacatgg agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc 721 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc 781 ttctcctggg acatcctgtc ccctcagttc atgtacggct ccagggcctt catcaagcac 841 cccgccgaca tccccgacta ctataagcag tccttccccg agggcttcaa gtgggagcgc 901 gtgatgaact tcgaggacgg cggcgccgtg accgtgaccc aggacacctc cctggaggac 961 ggcaccctga tctacaaggt gaagctccgc ggcaccaact tccctcctga cggccccgta 1021 atgcagaaga agacaatggg ctgggaagca tccaccgagc ggttgtaccc cgaggacggc 1081 gtgctgaagg gcgacattaa gatggccctg cgcctgaagg acggcggccg ctacctggcg 1141 gacttcaaga ccacctacaa ggccaagaag cccgtgcaga tgcccggcgc ctacaacgtc 1201 gaccgcaagt tggacatcac ctcccacaac gaggactaca ccgtggtgga acagtacgaa 1261 cgctccgagg gccgccactc caccggcggc atggacgagc tgtacaagtc cggactcaga 1321 tctcgagctc aagcttatac ctctaggaag aaagtgttgc tgaaggttat catcctggga 1381 gattctggag ttggtaagac atcactcatg aaccagtatg tgaacaagaa attcagtaat 1441 cagtacaaag ctacaatagg agcagacttt ctgacaaagg aggtgatggt ggatgacaga 1501 ctagttacaa tgcagatctg ggacacagca ggccaggaac ggttccagtc ccttggtgtg 1561 gccttctaca gaggtgcaga ctgctgcgtt ctggtatttg acgttactgc ccccaacaca 1621 ttcaaaaccc tcgatagctg gagagatgag tttctcatcc aggccagtcc ccgggatcct 1681 gaaaacttcc ctttcgttgt gttgggaaac aagattgacc tcgaaaacag acaagtggcc 1741 acaaagcggg cacaggcctg gtgctacagc aaaaacaaca ttccctactt cgagaccagt 1801 gccaaggagg ccatcaatgt ggagcaggcg ttccagacga ttgcaaggaa tgcacttaaa 1861 caggaaacag aggtggagct gtacaatgaa ttccctgaac ccatcaaact ggacaagaac 1921 gaccgggcca agacctcagc ggaaagctgc agttgctgaa agcttcgaat tctgcagtcg 1981 acggtaccgc gggcccggga tccaccggat ctagataact gatcataatc agccatacca 2041 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac 2101 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat 2161 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 2221 gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta 2281 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc 2341 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg 2401 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat 2461 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc 2521 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag 2581 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg 2641 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta 2701 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt 2761 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa 2821 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt 2881 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 2941 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3001 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 3061 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 3121 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg 3181 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt 3241 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat 3301 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag 3361 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac 3421 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac 3481 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc 3541 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg 3601 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag 3661 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat 3721 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag 3781 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc 3841 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg 3901 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg 3961 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag 4021 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat 4081 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 4141 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 4201 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga 4261 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg 4321 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg 4381 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg 4441 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga 4501 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc 4561 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa 4621 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 4681 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 4741 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt 4801 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 4861 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 4921 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 4981 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 5041 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 5101 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 5161 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 5221 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc 5281 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat //