LOCUS       sequence-493001-        5316 bp    DNA     circular SYN 18-NOV-2025
DEFINITION  Rab8 sensor donor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5316)
  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 5316)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 18, 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-Rab8a
FEATURES             Location/Qualifiers
     source          1..5316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    305..426
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(342..361)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     396..415
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(433..888)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(570..591)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     782..801
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        915..1019
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1021..1378
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1021..1212
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1044..1064)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1229..1364
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1413..2207
                     /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             1413..2207
                     /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(1467..1486)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2077..2096
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2395..2414)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2439..2486
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2815..3403
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3304..3323
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3578..3881
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3882..4085
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4036..4056
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4124..4133
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4130..4132,4133..4135,4136..4846)
                     /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(4175..4196)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4436..4455)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4783..4804
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 cagcgcgaag gccaacatca atgtggagaa cgcatttttc actctcgcca gggatatcaa
       61 agcaaaaatg gacaaaaaat tggaagggaa cagcccgcag gggagcagcc atggagtcaa
      121 gatcacagtg gagcagcaga agaggaccag cttcttccgg tgcagtctcc tgtgaggatc
      181 caccggatct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
      241 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
      301 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
      361 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
      421 atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
      481 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
      541 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
      601 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
      661 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
      721 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
      781 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
      841 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
      901 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
      961 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     1021 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     1081 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     1141 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     1201 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     1261 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     1321 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     1381 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     1441 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     1501 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     1561 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     1621 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     1681 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     1741 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     1801 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     1861 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     1921 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     1981 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     2041 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     2101 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     2161 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     2221 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     2281 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     2341 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     2401 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     2461 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     2521 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     2581 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     2641 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     2701 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2761 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2821 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2881 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2941 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3001 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3061 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3121 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3181 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3241 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3301 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3361 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3421 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3481 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
     3541 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     3601 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     3661 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     3721 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     3781 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     3841 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     3901 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     3961 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     4021 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     4081 gagctggttt agtgaaccgt cagatccgct agcgctaccg gtcgccacca tggtgagcaa
     4141 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     4201 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     4261 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     4321 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     4381 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     4441 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     4501 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     4561 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     4621 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     4681 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     4741 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     4801 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtccg gactcagatc
     4861 tcgagggatg gcgaagacct acgattacct gttcaagctg ctgctgatcg gggactcggg
     4921 ggtagggaag acctgtgtcc tgttccgctt ctccgaggac gccttcaatt ccacgttcat
     4981 ctctaccata ggaattgact ttaaaattag gaccatagag ctcgatggca agaggattaa
     5041 actgcagata tgggacacag ccggccagga gcggtttcgg acaatcacga cagcctacta
     5101 caggggtgcc atgggtatca tgctggtcta cgacattacc aatgagaagt ccttcgacaa
     5161 catccggaat tggattcgga acattgagga gcatgcctct gcagatgtgg agaagatgat
     5221 actggggaat aagtgtgatg tgaacgacaa gaggcaggtg tccaaggaac ggggagaaaa
     5281 gctggcactc gactatggga tcaagttcat ggagac
//
