LOCUS       sequence-488620-        6164 bp    DNA     circular SYN 07-OCT-2025
DEFINITION  Rab4a (Dominant negative).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6164)
  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 6164)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 7, 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: pCAGGS-mEGFP-Rab4a(S27N)
FEATURES             Location/Qualifiers
     source          1..6164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        19..398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        400..677
                     /label=chicken beta-actin promoter
     intron          678..1698
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1618..1640
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1706..1725
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1769..1778
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1775..1777,1778..1780,1781..2491)
                     /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(1820..1841)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2081..2100)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2428..2449
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3196..3215)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3261..3316
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3315..3334)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3677..3693)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3677..3693)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3690..3712)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3701..3717
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3725..3731,3732..3749,3750..3755))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3770..3791
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3850..4045
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3896..4031
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4051..4185
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4088..4107)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4142..4161
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4253..4270)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4424..5012)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4504..4523)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5183..5974,5975..6043))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5806..5825
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6044..6148)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg ggtcgatcga cattgattat tgactagtta ttaatagtaa tcaattacgg
       61 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      121 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      181 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      241 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      301 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      361 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct
      421 gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt
      481 aattattttg tgcagcgatg ggggcggggg gggggggggg gcgcgcgcca ggcggggcgg
      541 ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg
      601 gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc
      661 gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg
      721 ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc
      781 gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt
      841 ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg
      901 gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc
      961 ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg
     1021 aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag
     1081 gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc
     1141 tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg
     1201 ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg
     1261 gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg
     1321 gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt
     1381 aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg
     1441 ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga
     1501 aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc
     1561 agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt
     1621 tcggcttctg gcgtgtgacc ggcggctcta gctagagcct ctgctaacca tgttcatgcc
     1681 ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt
     1741 ggcaaagaat tcgctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt
     1801 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1861 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1921 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     1981 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     2041 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     2101 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     2161 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     2221 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     2281 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     2341 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag
     2401 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     2461 gatcactctc ggcatggacg agctgtacaa gtccggactc agatctcgag ggatggcgca
     2521 gaccgccatg tccgagactt acgatttctt gtttaagttc ttggtcattg gaaatgcggg
     2581 aactggcaaa aactgcttgc tccatcagtt cattgagaag aaattcaaag atgactcaaa
     2641 tcataccata ggagtggaat tcggctcaaa gataataaat gttggtggta aatatgtgaa
     2701 gttacagata tgggacacgg ctggacagga gcggttcagg tctgtgacga gaagctacta
     2761 cagaggtgcg gctggggcac tcctcgtcta tgacatcacc agccgagaaa cctacaatgc
     2821 gcttactaat tggttaacag atgccagaat gctggcgagc cagaacatcg tcatcattct
     2881 ctgcgggaac aagaaggacc tggatgccga ccgggaagtc accttccttg aagcctccag
     2941 gttcgcacaa gagaatgagc tcatgttcct ggaaaccagt gcactgactg gcgagaacgt
     3001 cgaagaggct ttcatgcagt gcgcaaggaa gatacttaac aaaattgaat caggtgagct
     3061 ggaccccgag aggatgggct ctggtatcca gtatggagac gccgccttga gacagctacg
     3121 gtcaccccga cgtacacagg ctccaagtgc acaggagtgt ggctgctagg cggccgcgac
     3181 tctagagtcg acggatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct
     3241 tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     3301 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa
     3361 tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg
     3421 ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca
     3481 tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc
     3541 tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg
     3601 actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa
     3661 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3721 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     3781 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3841 agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3901 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3961 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     4021 ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac
     4081 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4141 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa
     4201 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4261 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4321 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4381 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4441 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4501 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4561 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4621 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4681 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4741 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4801 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4861 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4921 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4981 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5041 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5101 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5161 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5221 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5281 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5341 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5401 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5461 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5521 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5581 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5641 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5701 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5761 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5821 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5881 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5941 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6001 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6061 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6121 agaaaaataa acaaataggg gttccgcgca catttccccg aaaa
//
