LOCUS       sequence-197924-        5382 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Point mutation in Rab5 that reestablishes interaction with APPL1 
            when co-expressed with APPL1-N308D.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5382)
  TITLE     Webb Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5382)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: Rab5-L38R-mCherry
FEATURES             Location/Qualifiers
     source          1..5382
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     50..70
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(95..199)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      226..681
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(313..332)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     523..544
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    688..809
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(699..718)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     753..772
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1653..2360)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1928..1947)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2197..2215
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2357..2366
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2411..2614)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2440..2460)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2615..2918
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3093..3681)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3173..3192)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4010..4057
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4082..4101
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4289..5083)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4289..5083)
                     /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(4400..4419)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5010..5029
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5118..5382,1..93))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5132..5267
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        join(5284..5382,1..93)
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
ORIGIN
        1 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga
       61 cacacattcc acagctggtt ctttccgcct caggactctt cctttttcaa tattattgaa
      121 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
      181 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg
      241 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
      301 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
      361 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
      421 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
      481 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
      541 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
      601 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
      661 aaatattaac gcttacaatt tacgcgttaa gatacattga tgagtttgga caaaccacaa
      721 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
      781 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
      841 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
      901 tggctgatta tgatcagtta tctagatccg gtggatcccg ggcccgcggt accgtcgact
      961 tagttactac aacactgatt cctggttggt tgtgtgggtt cggtaaggtc tactcctctt
     1021 cctctggcag aatttgctcc tggattttgt ggttcattct ttggcaattt tttagctatt
     1081 gccatgaata tttcatttac attcattgat gttttagcgg atgtctccat gaataataaa
     1141 ctattgtcat ctgcatagga ctgtgcttcc tggaaatcta ctgctctttt atttgctagg
     1201 tcggccttgt ttcccgataa agctattaca atgttaggac ttgcttgcct ctgaagttct
     1261 ttaacccaat tttttgctct tgcaaaggac tcctcatttg tgatatcata tacaactatg
     1321 gctgcttgtg ctcctctgta gtacattggt gctaggctat ggtatcgttc ttgaccagct
     1381 gtatcccata tttcaaactt tactgtagtg tcatcaagac atacagtttg ggttagaaaa
     1441 gcagccccaa tggtactctc ttgaaattca tgaaattggc ctttcacaaa acgacgcact
     1501 aggcttgatt tgccaacagc ggactctccc agaagtacta gtttgaactg gcatatttta
     1561 tttccagtat ttggcccgtt gggtcttgtt gcgcctcgac tagccatgaa ttcagccgag
     1621 ttcgaagctt gtgctcgaga tctgagtccg gacttgtaca gctcgtccat gccgccggtg
     1681 gagtggcggc cctcggcgcg ttcgtactgt tccacgatgg tgtagtcctc gttgtgggag
     1741 gtgatgtcca acttgatgtt gacgttgtag gcgccgggca gctgcacggg cttcttggcc
     1801 ttgtaggtgg tcttgacctc agcgtcgtag tggccgccgt ccttcagctt cagcctctgc
     1861 ttgatctcgc ccttcagggc gccgtcctcg gggtacatcc gctcggagga ggcctcccag
     1921 cccatggtct tcttctgcat tacggggccg tcggagggga agttggtgcc gcgcagcttc
     1981 accttgtaga tgaactcgcc gtcctgcagg gaggagtcct gggtcacggt caccacgccg
     2041 ccgtcctcga agttcatcac gcgctcccac ttgaagccct cggggaagga cagcttcaag
     2101 tagtcgggga tgtcggcggg gtgcttcacg taggccttgg agccgtacat gaactgaggg
     2161 gacaggatgt cccaggcgaa gggcaggggg ccacccttgg tcaccttcag cttggcggtc
     2221 tgggtgccct cgtaggggcg gccctcgccc tcgccctcga tctcgaactc gtggccgttc
     2281 acggagccct ccatgtgcac cttgaagcgc atgaactcct tgatgatggc catgttatcc
     2341 tcctcgccct tgctcaccat ggtggcaagc ttaagtttaa acgctagcgg atctgacggt
     2401 tcactaaacc agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg
     2461 tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca
     2521 aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc
     2581 cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt
     2641 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg
     2701 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta
     2761 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
     2821 ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg
     2881 gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat
     2941 gaactaatga ccccgtaatt gattactatt aataactaat gcatggcggt aatacggtta
     3001 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3061 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3121 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3181 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3241 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3301 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3361 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3421 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3481 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3541 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3601 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3661 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3721 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3781 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaacct
     3841 gaggctatgg cagggcctgc cgccccgacg ttggctgcga gccctgggcc ttcacccgaa
     3901 cttggggggt ggggtgggga aaaggaagaa acgcgggcgt attggcccca atggggtctc
     3961 ggtggggtat cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac
     4021 ccaacaccgt gcgttttatt ctgtcttttt attgccgtca tagcgcgggt tccttccggt
     4081 attgtctcct tccgtgtttc agttagcctc cccctagggt gggcgaagaa ctccagcatg
     4141 agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac
     4201 ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg
     4261 tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg
     4321 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
     4381 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg
     4441 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
     4501 tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct
     4561 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
     4621 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt
     4681 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
     4741 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc
     4801 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa
     4861 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac
     4921 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg
     4981 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
     5041 aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc
     5101 ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
     5161 tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
     5221 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
     5281 ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     5341 gagcctgggg actttccaca cctggttgct gactaattga ga
//
