LOCUS       sequence-197993-        6864 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Point mutation in APPL1 that diminishes its interaction with Rab5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6864)
  TITLE     Webb Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6864)
  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            "Unpublished"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: APPL1-N308D-mCherry
FEATURES             Location/Qualifiers
     source          1..6864
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        43..234
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     191..211
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(236..340)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      367..822
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(454..473)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     664..685
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    829..950
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(840..859)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     894..913
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3282..3989)
                     /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(3557..3576)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3826..3844
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3986..3995
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4034..4237)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4063..4083)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4238..4541
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(4716..5304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4796..4815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5633..5680
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5705..5724
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5912..6706)
                     /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(5912..6706)
                     /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(6023..6042)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6633..6652
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6741..6864,1..234))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6755..6864,1..26)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
       61 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
      121 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
      181 gactttccac accctaactg acacacattc cacagctggt tctttccgcc tcaggactct
      241 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
      301 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
      361 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
      421 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      481 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      541 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      601 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      661 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      721 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      781 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttacgcgtta agatacattg
      841 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
      901 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
      961 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1021 aaaacctcta caaatgtggt atggctgatt atgatcagtt atctagatcc ggtggatccc
     1081 gggcccgcgg taccgtcgac tgcagaattc ggctttgctt ctgattctct cttctttcct
     1141 ccttctccca aatcactctc ttctgactgg ctactgctaa ggacaacaaa ctgcccctca
     1201 ctactggctt ggtttggtct actggaagca gctatcaacc gactttgttc ttccaagtcc
     1261 ttttcaatct gtttttgttt attgagttct ttctgttgct tctcttttac tctctctatt
     1321 tctttttgtt tttctgatgc cctacgatcc agttcagcat gcaaagctat ctgttttgcc
     1381 agtccaacag aatcacatat cttttccccc tcattgtttg actcaaatat atagcagact
     1441 gatgacagat tactttcact tctcccgctt gatgtccgaa gaacaaatcc aaaaaggcgc
     1501 ttattttcct ggtgtgtagc atacaaaact acacaaggta atggaaacgt gagccttgta
     1561 acttgtgtct gtggatcaat taactttaaa cagtcacaag tgactaataa atgcgattct
     1621 gtcatacgaa agatgttatg gatggcccgg gcagctaaga tttggcgcat agtttcataa
     1681 acaacatctg gatggtcatc tgatttcacc tccattgaac caaggaatcg gacaataaat
     1741 aactgatgaa gaatagaatc ttcagtttca gattttgtac ttcctccaga ttctccaaat
     1801 ggatttgtac gcctgcctcc ctggccaaag gcttttgcct ggccaggctg atcttcacac
     1861 acaggagaaa ttatgtcaaa ctgtattggg gtgtctgggg caacaagaga atctagagag
     1921 agggcagcca aatttgtaga ctcagatcct aaggatcctg aactgctggt tcgagctgtc
     1981 ggtggccgag attgtcctgc tgctggccgc aggctctcgt gcctctgctg gaaagatggg
     2041 gaaggagtga cagcttccag agctgattga tttactcgtg cagcagtttc ctctggattt
     2101 tcacttaagt atatttgttt agatatgttg tttattgtac agatccactc ttcatgatct
     2161 tttttactct ctgcttgcaa aattgaagat ttttttccat cgaaagaggt gatctgaaaa
     2221 caatatcgtc tgtcttcaca gtccacagcc atcactgaac agttgtctat gtccatggcc
     2281 aggcctcctg ctacatcccc acgggcctga ctcattaaat ctccaccctg cgtgaagtaa
     2341 aactgtctgt cccaggtaga tgacaccaag cctgttttat tcctagcatt aaggtatcca
     2401 gcctttcggg ttaaatttcg attaacagga aatttggtgg ggtctgggtc aggcacatat
     2461 aagggatcac tggctacttc caaatcctct attgtctgtt gcatggtctc tatatcactg
     2521 tccatttccc tgcgaacatt ctgaacgctt gttccaatat tagctaaaaa ttcttccagt
     2581 tgttcattaa gattttcaga acccatctta aagaaactta tctgagcttg catgtaccca
     2641 agtagaggtt ctaacaatgc tattttcttc ttgtactgaa gagtatttaa tgcacaaaaa
     2701 taatgcatca tggtctggtg ttgtttcttt ctggatgtgt acacatcttc tgttacttca
     2761 tacttcacct tgtcattttc tctttttttt gataaacggc tatatctatt aatcgcagca
     2821 tcatgatcat tacttgcaat ctgaaatact tcctttaatg ttagtatttc tttcagatct
     2881 ctttctttaa actgggtaat ggggaacatc atggcatcag caagttgagt tgaaagcact
     2941 gcatgacaag agctaagctc atctataact tttgaaaact gttgcaatgt agagctcata
     3001 acttcatcat cacctcccaa tggaaaacgc tgtttttcat attcttttaa aagttttgag
     3061 gtcaggtgtg ttgctgcact taattcattc tgtgcatcat aaatccgatg catagcttga
     3121 tacaactggt tcatatagtt ggaaatagct gtggcatctt cttcaaatac acctagtaaa
     3181 gaccttgtct gcgggctgtc ctccagcgtc tcctcgatgg gcagcttgtc gatccccggc
     3241 ataagccgaa ttcgaagctt gagctcgaga tctgagtccg gcttgtacag ctcgtccatg
     3301 ccgccggtgg agtggcggcc ctcggcgcgt tcgtactgtt ccacgatggt gtagtcctcg
     3361 ttgtgggagg tgatgtccaa cttgatgttg acgttgtagg cgccgggcag ctgcacgggc
     3421 ttcttggcct tgtaggtggt cttgacctca gcgtcgtagt ggccgccgtc cttcagcttc
     3481 agcctctgct tgatctcgcc cttcagggcg ccgtcctcgg ggtacatccg ctcggaggag
     3541 gcctcccagc ccatggtctt cttctgcatt acggggccgt cggaggggaa gttggtgccg
     3601 cgcagcttca ccttgtagat gaactcgccg tcctgcaggg aggagtcctg ggtcacggtc
     3661 accacgccgc cgtcctcgaa gttcatcacg cgctcccact tgaagccctc ggggaaggac
     3721 agcttcaagt agtcggggat gtcggcgggg tgcttcacgt aggccttgga gccgtacatg
     3781 aactgagggg acaggatgtc ccaggcgaag ggcagggggc cacccttggt caccttcagc
     3841 ttggcggtct gggtgccctc gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg
     3901 tggccgttca cggagccctc catgtgcacc ttgaagcgca tgaactcctt gatgatggcc
     3961 atgttatcct cctcgccctt gctcaccatg gtggcgaccg gtagcgctag cggatctgac
     4021 ggttcactaa accagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     4081 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     4141 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     4201 atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata
     4261 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     4321 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     4381 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     4441 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     4501 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     4561 tatgaactaa tgaccccgta attgattact attaataact aatgcatggc ggtaatacgg
     4621 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4681 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4741 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4801 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4861 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4921 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4981 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5041 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5101 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5161 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5221 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5281 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5341 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5401 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5461 cctgaggcta tggcagggcc tgccgccccg acgttggctg cgagccctgg gccttcaccc
     5521 gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt
     5581 ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac
     5641 gacccaacac cgtgcgtttt attctgtctt tttattgccg tcatagcgcg ggttccttcc
     5701 ggtattgtct ccttccgtgt ttcagttagc ctccccctag ggtgggcgaa gaactccagc
     5761 atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat tccgaagccc
     5821 aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct
     5881 tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga
     5941 aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc
     6001 attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt
     6061 ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga
     6121 tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg
     6181 ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat
     6241 cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt
     6301 ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca
     6361 tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt
     6421 cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag
     6481 gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg
     6541 caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca
     6601 cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca
     6661 cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc
     6721 atcctgtctc ttgatcgatc tttgcaaaag cctaggcctc caaaaaagcc tcctcactac
     6781 ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt
     6841 cagccatggg gcggagaatg ggcg
//
