LOCUS       sequence-489525-        5357 bp    DNA     circular SYN 16-OCT-2025
DEFINITION  Rab5a (Dominant negative).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5357)
  AUTHORS   Wang J, Nishiyama J, Parra-Bueno P, Okaz E, Oz G, Liu X, Watabe T, 
            Suponitsky-Kroyter I, McGraw TE, Szatmari EM, Yasuda R
  TITLE     Rab10 inactivation promotes AMPAR trafficking and spine enlargement 
            during long-term potentiation.
  JOURNAL   Elife. 2025 Sep 23;13:RP103879. doi: 10.7554/eLife.103879.
  PUBMED    40986440
REFERENCE   2  (bases 1 to 5357)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2025-09-23"; volume: "13:RP103879. doi"; pages: " 
            10.7554/eLife.103879"
COMMENT     Sequence Label: CMV-mEGFP-Rab5a(S34N)
FEATURES             Location/Qualifiers
     source          1..5357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        33..336
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        337..540
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     491..511
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      579..588
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(585..587,588..590,591..1301)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(630..651)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(891..910)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1238..1259
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2117..2238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2154..2173)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2208..2227
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2245..2700)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2382..2403)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2594..2613
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2727..2831
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2833..3190
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2833..3024
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2856..2876)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3041..3176
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3225..4019
                     /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             3225..4019
                     /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(3279..3298)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3889..3908
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4207..4226)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4251..4298
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4627..5215
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5116..5135
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
       61 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      121 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      181 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      241 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      301 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      361 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      421 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      481 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      541 ggtttagtga accgtcagat ccgctagcgc taccggtcgc caccatggtg agcaagggcg
      601 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      661 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      721 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
      781 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
      841 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
      901 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
      961 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     1021 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     1081 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     1141 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     1201 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1261 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtccggactc agatctcgag
     1321 ctcgggctaa tcgaggagca acaagaccca acgggccaaa tactggaaat aaaatatgcc
     1381 agttcaaact ggttcttcta ggagagtctg ctgttggcaa aaacagtctg gtgcttcgct
     1441 ttgtgaaagg ccaatttcat gagtttcaag agagtaccat tggggctgcc tttctaaccc
     1501 aaactgtgtg tcttgatgac acaacagtaa aatttgaaat atgggataca gctggccaag
     1561 aacggtatca tagcctagca ccaatgtact accgaggagc acaagccgcc atagttgtgt
     1621 atgatattac aaatgaggag tccttttcga gagcaaaaaa ctgggttaaa gaacttcaaa
     1681 ggcaagcaag tcctaatatt gtgatagctt tgtcaggaaa caaggctgac ttagcaaata
     1741 aaagagctgt tgacttccag gaagcacagt cctatgcaga tgacaacagc ttattattta
     1801 tggagacatc agctaagaca tcaatgaatg taaatgaaat atttatggca atagctaaaa
     1861 agctgccaaa gaacgaacca caaaatccag gcgcaaactc agccagagga cgaggagtag
     1921 accttactga gcctgcacag ccagccagaa gccagtgttg tagtaactaa ggatccgcgg
     1981 ccgctaggga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
     2041 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     2101 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     2161 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2221 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     2281 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     2341 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     2401 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     2461 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     2521 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     2581 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     2641 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     2701 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2761 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     2821 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     2881 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2941 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3001 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3061 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3121 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3181 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3241 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3301 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3361 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     3421 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3481 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3541 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3601 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3661 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3721 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     3781 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     3841 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     3901 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     3961 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4021 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4081 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4141 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     4201 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     4261 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     4321 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     4381 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     4441 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     4501 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4561 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4621 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4681 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4741 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4801 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4861 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4921 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4981 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5041 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5101 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5161 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5221 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5281 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat
     5341 agtaatcaat tacgggg
//
