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 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..1287 /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(743..761) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1011..1030 /label=mCherry-F /note="mCherry, forward primer" primer_bind 1205..1228 /label=DsRed1-C /note="DsRed1, forward primer" polyA_signal 2109..2230 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2146..2165) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2200..2219 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2237..2692) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2374..2395) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2586..2605 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2719..2823 /gene="bla" /label=AmpR promoter promoter 2825..3182 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2848..2868) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3033..3168 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3095..3114 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3217..4011 /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(3271..3290) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3881..3900 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4199..4218) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4243..4290 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4619..5207 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5108..5127 /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 ccatggcctc ctccgaggac gtcatcaagg agttcatgcg cttcaaggtg 661 cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc 721 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc 781 gcctgggaca tcctgtcccc tcagttccag tacggctcca aggcctacgt gaagcacccc 841 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg 901 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc 961 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg 1021 cagaagaaga ccatgggctg ggaggcctcc accgagcgga tgtaccccga ggacggcgcc 1081 ctgaagggcg agatcaagat gaggctgaag ctgaaggacg gcggccacta cgacgccgag 1141 gtcaagacca cctacatggc caagaagccc gtgcagctgc ccggcgccta caagaccgac 1201 atcaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc 1261 gccgagggcc gccactccac cggcgccctg tacaagtact cagatctcga gatgacctct 1321 aggaagaaag tgttgctgaa ggttatcatc ctgggagatt ctggagttgg taagacatca 1381 ctcatgaacc agtatgtgaa caagaaattc agtaatcagt acaaagctac aataggagca 1441 gactttctga caaaggaggt gatggtggat gacagactag ttacaatgca gatctgggac 1501 acagcaggcc aggaacggtt ccagtccctt ggtgtggcct tctacagagg tgcagactgc 1561 tgcgttctgg tatttgacgt tactgccccc aacacattca aaaccctcga tagctggaga 1621 gatgagtttc tcatccaggc cagtccccgg gatcctgaaa acttcccttt cgttgtgttg 1681 ggaaacaaga ttgacctcga aaacagacaa gtggccacaa agcgggcaca ggcctggtgc 1741 tacagcaaaa acaacattcc ctacttcgag accagtgcca aggaggccat caatgtggag 1801 caggcgttcc agacgattgc aaggaatgca cttaaacagg aaacagaggt ggagctgtac 1861 aatgaattcc ctgaacccat caaactggac aagaacgacc gggccaagac ctccgcagaa 1921 agctgcagtt gttaaatcga taagcttcga attctgcagt cgacggtacc gcgggcccgg 1981 gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac 2041 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg 2101 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 2161 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 2221 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt 2281 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca 2341 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta 2401 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta 2461 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg 2521 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga 2581 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg 2641 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc 2701 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat 2761 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag 2821 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc 2881 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt 2941 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 3001 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 3061 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 3121 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 3181 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac 3241 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca 3301 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt 3361 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg 3421 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga 3481 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct 3541 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg 3601 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg 3661 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc 3721 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat 3781 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac 3841 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt 3901 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct 3961 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc 4021 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca 4081 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga 4141 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg 4201 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga 4261 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc 4321 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt 4381 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg 4441 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt 4501 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt 4561 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt 4621 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag 4681 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca 4741 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca 4801 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 4861 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 4921 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 4981 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 5041 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 5101 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 5161 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 5221 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt 5281 tatcccctga ttctgtggat aaccgtatta ccgccatgca t //