LOCUS       Exported                5319 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression of Rab5 fused to mRFP. Used for localization to
            the early endosome, and other early endosomal vacuoles..
ACCESSION   .
VERSION     .
KEYWORDS    mRFP-Rab5
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5319)
  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 5319)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5319
                     /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    2107..2228
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2144..2163)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2198..2217
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2235..2690)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2372..2393)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2584..2603
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2717..2821
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2823..3180
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2846..2866)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3031..3166
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3093..3112
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3215..4009
                     /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(3269..3288)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3879..3898
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4197..4216)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4241..4288
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4617..5205
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5106..5125
                     /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 gctcaagctt
     1321 cgaattccga tggctagtcg aggcgcaaca agacccaacg ggccaaatac tggaaataaa
     1381 atatgccagt tcaaactagt acttctggga gagtccgctg ttggcaaatc aagcctagtg
     1441 cttcgttttg tgaaaggcca atttcatgaa tttcaagaga gtaccattgg ggctgctttt
     1501 ctaacccaaa ctgtatgtct tgatgacact acagtaaagt ttgaaatatg ggatacagct
     1561 ggtcaagaac gataccatag cctagcacca atgtactaca gaggagcaca agcagccata
     1621 gttgtatatg atatcacaaa tgaggagtcc tttgcaagag caaaaaattg ggttaaagaa
     1681 cttcagaggc aagcaagtcc taacattgta atagctttat cgggaaacaa ggccgaccta
     1741 gcaaataaaa gagcagtaga tttccaggaa gcacagtcct atgcagatga caatagttta
     1801 ttattcatgg agacatccgc taaaacatca atgaatgtaa atgaaatatt catggcaata
     1861 gctaaaaaat tgccaaagaa tgaaccacaa aatccaggag caaattctgc cagaggaaga
     1921 ggagtagacc ttaccgaacc cacacaacca accaggaatc agtgttgtag taactaagga
     1981 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     2041 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     2101 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2161 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2221 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     2281 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     2341 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     2401 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     2461 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     2521 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     2581 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     2641 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     2701 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2761 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2821 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     2881 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     2941 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3001 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3061 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3121 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3181 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     3241 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     3301 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     3361 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     3421 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     3481 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     3541 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     3601 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     3661 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     3721 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     3781 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     3841 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     3901 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     3961 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4021 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4081 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4141 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     4201 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     4261 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     4321 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     4381 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     4441 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     4501 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4561 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4621 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4681 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4741 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4801 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4861 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4921 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4981 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5041 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5101 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5161 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5221 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5281 tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
