LOCUS       sequence-488619-        6120 bp    DNA     circular SYN 07-OCT-2025
DEFINITION  Rab7a (Constitutively active).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6120)
  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 6120)
  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-Rab7a(Q67L)
FEATURES             Location/Qualifiers
     source          1..6120
                     /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(3152..3171)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3217..3272
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3271..3290)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3633..3649)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3633..3649)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3646..3668)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3657..3673
                     /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(3681..3687,3688..3705,3706..3711))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3726..3747
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3806..4001
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3852..3987
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4007..4141
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4044..4063)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4098..4117
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4209..4226)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4380..4968)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4460..4479)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5139..5930,5931..5999))
                     /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     5762..5781
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6000..6104)
                     /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 gcatgacctc
     2521 taggaagaaa gtgttgctga aggtcatcat cctgggagac tctggtgttg gaaagacatc
     2581 gctcatgaac cagtatgtga ataagaagtt cagtaaccag tacaaagcca caataggagc
     2641 agactttctg accaaggagg tgatggtgga tgacagactt gttaccatgc agatctggga
     2701 cacagctggt ctagaacggt tccagtctct tggtgtggcc ttctacagag gtgcagattg
     2761 ctgcgttctg gtgtttgatg tgactgcccc caacacattc aaaacccttg acagctggag
     2821 agacgagttt ctcatccagg ccagcccccg ggatcctgag aacttccctt ttgttgtgtt
     2881 gggaaacaag attgacctgg aaaacagaca agtggccaca aagagggcac aggcttggtg
     2941 ctacagcaaa aacaacattc cttacttcga gaccagtgcc aaggaggcca tcaatgtgga
     3001 acaggccttc cagacaattg ctcggaatgc ccttaaacag gaaacggaag tggagctgta
     3061 caatgaattc cctgaaccca tcaaactgga caagaacgag cgggccaagg cctccgcaga
     3121 gagctgcagc tgttgatcta gagtcgacgg atctttttcc ctctgccaaa aattatgggg
     3181 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     3241 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     3301 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     3361 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     3421 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     3481 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     3541 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc
     3601 tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     3661 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3721 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3781 ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc
     3841 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3901 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3961 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc
     4021 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4081 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     4141 ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4201 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4261 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4321 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4381 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4441 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4501 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4561 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4621 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4681 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4741 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4801 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4861 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4921 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4981 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5041 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5101 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5161 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5221 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5281 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5341 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5401 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5461 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5521 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5581 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5641 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5701 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5761 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5821 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5881 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5941 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6001 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6061 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
//
