LOCUS Exported 5321 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression of Rab7 fused to mRFP. Used for localization to the late endosome, and other early endosomal vacuoles.. ACCESSION . VERSION . KEYWORDS mRFP-Rab7 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5321) AUTHORS Vonderheit A, Helenius A TITLE Rab7 associates with early endosomes to mediate sorting and transport of Semliki forest virus to late endosomes. JOURNAL PLoS Biol. 2005 Jul . 3(7):e233. PUBMED 15954801 REFERENCE 2 (bases 1 to 5321) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5321 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 119..422 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 423..626 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 577..597 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 665..674 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 671..1345 /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /note="mammalian codon-optimized" /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS TGA" primer_bind complement(801..819) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1069..1088 /label=mCherry-F /note="mCherry, forward primer" primer_bind 1263..1286 /label=DsRed1-C /note="DsRed1, forward primer" polyA_signal 2167..2288 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2204..2223) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2258..2277 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2295..2750) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2432..2453) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2644..2663 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2777..2881 /gene="bla" /label=AmpR promoter promoter 2883..3240 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2906..2926) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3091..3226 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3153..3172 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3275..4069 /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(3329..3348) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3939..3958 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4257..4276) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4301..4348 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4677..5265 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5166..5185 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta 61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat 481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac 601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc 661 ggtcgccacc atggcctcct ccgaggacgt catcaaggag ttcatgcgct tcaaggtgcg 721 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc 781 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc 841 ctgggacatc ctgtcccctc agttccagta cggctccaag gcctacgtga agcaccccgc 901 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat 961 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga 1021 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca 1081 gaagaagacc atgggctggg aggcctccac cgagcggatg taccccgagg acggcgccct 1141 gaagggcgag atcaagatga ggctgaagct gaaggacggc ggccactacg acgccgaggt 1201 caagaccacc tacatggcca agaagcccgt gcagctgccc ggcgcctaca agaccgacat 1261 caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgagcgcgc 1321 cgagggccgc cactccaccg gcgccctgta caagtactca gatctcgaga tgacctctag 1381 gaagaaagtg ttgctgaagg ttatcatcct gggagattct ggagttggta agacatcact 1441 catgaaccag tatgtgaaca agaaattcag taatcagtac aaagctacaa taggagcaga 1501 ctttctgaca aaggaggtga tggtggatga cagactagtt acaatgcaga tctgggacac 1561 agcaggccag gaacggttcc agtcccttgg tgtggccttc tacagaggtg cagactgctg 1621 cgttctggta tttgacgtta ctgcccccaa cacattcaaa accctcgata gctggagaga 1681 tgagtttctc atccaggcca gtccccggga tcctgaaaac ttccctttcg ttgtgttggg 1741 aaacaagatt gacctcgaaa acagacaagt ggccacaaag cgggcacagg cctggtgcta 1801 cagcaaaaac aacattccct acttcgagac cagtgccaag gaggccatca atgtggagca 1861 ggcgttccag acgattgcaa ggaatgcact taaacaggaa acagaggtgg agctgtacaa 1921 tgaattccct gaacccatca aactggacaa gaacgaccgg gccaagacct ccgcagaaag 1981 ctgcagttgt taaatcgata agcttcgaat tctgcagtcg acggtaccgc gggcccggga 2041 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt 2101 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt 2161 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 2221 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 2281 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg 2341 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa 2401 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa 2461 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg 2521 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa 2581 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa 2641 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct 2701 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac 2761 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat 2821 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag 2881 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca 2941 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt 3001 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 3061 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 3121 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 3181 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa 3241 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc 3301 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 3361 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 3421 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg 3481 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 3541 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc 3601 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 3661 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 3721 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 3781 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg 3841 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 3901 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 3961 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 4021 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg 4081 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc 4141 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 4201 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa 4261 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat 4321 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac 4381 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc 4441 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca 4501 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt 4561 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 4621 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 4681 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 4741 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 4801 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag 4861 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 4921 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 4981 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 5041 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 5101 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 5161 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 5221 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 5281 gcctttttac ggttcctggc cttttgctgg ccttttgctc a //