LOCUS       sequence-488603-        7217 bp    DNA     circular SYN 07-OCT-2025
DEFINITION  Rab5 GAP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7217)
  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 7217)
  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-USP6NL
FEATURES             Location/Qualifiers
     source          1..7217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        19..398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        400..675
                     /label=chicken beta-actin promoter
     intron          676..1696
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1616..1638
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1704..1723
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3269..3280
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4249..4268)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4314..4369
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4368..4387)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4730..4746)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4730..4746)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4743..4765)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4754..4770
                     /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(4778..4784,4785..4802,4803..4808))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4823..4844
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4903..5098
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4949..5084
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5104..5238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5141..5160)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5195..5214
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5306..5323)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5477..6065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5557..5576)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6236..7027,7028..7096))
                     /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     6859..6878
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7097..7201)
                     /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 gggggggggc gcgcgccagg cggggcgggg
      541 cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc
      601 gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga
      661 agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc
      721 gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg
      781 gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt
      841 ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc
      901 tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc
      961 ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag
     1021 gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc
     1081 tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg
     1141 caaccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg
     1201 ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg
     1261 gtgccgggcg gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc
     1321 ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa
     1381 tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg
     1441 aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag
     1501 gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag
     1561 cctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc
     1621 ggcttctggc gtgtgaccgg cggctctagc tagagcctct gctaaccatg ttcatgcctt
     1681 cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg
     1741 caaagaattc gctagcgcta ccggtgccac catgaattcg gaccaggatg tagcgctcaa
     1801 actcgctcag gaacgagctg aaatagttgc taaatatgac agaggacggg aaggtgcaga
     1861 gatcgagccc tgggaagatg ctgattatct tgtttacaaa gtcaccgata gatttggctt
     1921 tttacatgag gaggagctcc cgtatcctaa cgcagccgcg gaccggcaaa agcaactgga
     1981 aatcgaaagg acttccaagt ggctgaaaat gctgaagaga tgggagagat acaagaacac
     2041 tgagaagttc catcgacgaa tttacaaagg aattccactc cagctccgag gtgaagtctg
     2101 ggcccttctt cttgagatcc ctaaaatgaa agaagaaaca agagacctct atagtaaatt
     2161 aaagcacaga gcacggggtt gttcgcctga tatcagacaa atagaccttg acgtgaaccg
     2221 cacattcagg gaccatatca tgtttagaga cagatatggt gtcaagcagc aatccttgtt
     2281 ccacgttctt gctgcctact ccatctacaa tacggaggtc gggtactgtc aagggatgag
     2341 tcagatcaca gctctgctcc tgatgtacat gaatgaggag gacgccttct gggccctggt
     2401 caagctcttc tcaggcccca agcacgccat gcatggtttt tttgtccaag gtttccctaa
     2461 actcttgaga tttcaagaac atcatgagaa aattctgaac aaatttttgt ccaaactaaa
     2521 gcagcacttg gattctcaag aaatctatac aagtttttac acaatgaagt ggttttttca
     2581 gtgtttcctt gaccgaactc ccttcaggct gaacctccgg atctgggaca tctacatctt
     2641 tgagggagag cgaattctca ctgctatgtc ttatacaatc ttgaagctac acagaaagca
     2701 tctaatgaag ttgtctatgg aagaacttgt agaatttctt caggagactc tcgccaagga
     2761 ctttttcttc gaggatgatt tcgtaataga gcaactgcag gtctctatga cagagttaaa
     2821 gcgggccaag ttagatctcc cagaacctgg taaggaggat gaatacccaa ggaaacccct
     2881 ggggcagctc ccacctgagt ataccaatca cttgagcaat ggacaaagaa gtgttggcag
     2941 gcccagcccc aaaataagca gcaggagaga agatgggtcg cctcacaagg cacatgagca
     3001 ctctcctgcc catcacagca gaaatggcac cccagaaaga gtcgggcagt caagacgcaa
     3061 gtctgtggat gagggtagta aaaatctgaa gcaggaggca gattttcaga gaaaaccttc
     3121 atcgggcaca caggacaact ccaagcatta taatcatgca actgctaacc aaaatagcaa
     3181 tgcactttcc aacatcagga aggagttcgt gcccaaatgg aggaagccat cggaggcctc
     3241 agctatcgaa agaactgcta aatacgccat tgaaggcaga agtcactcag tgtttcccgc
     3301 tctctctgtc gcagtcccaa gttctgctga gacccgattg tcaaactcaa ggcaaaagat
     3361 gaaggctttt gatggtggtg aggggaaccg gggttctaat gcttcccagt atgacaacgt
     3421 acctgggagt gagagcgagc ctggagcgtc tgcagaggag ggaccggagc ggactcaccc
     3481 tcacagtcca aggaagcact ctgagccaag tcccagtcca tcaaaagtac ccaacaagtt
     3541 tacctttaaa gtgcagcctc ccagccatgt acgataccca tcccagccag caggggagga
     3601 tcacagagct gcatatccac cctcctacag caacccgcct gtctaccatg gaaattctcc
     3661 aaaacacatc cctacggccc acagtggctt tgcgtccaca cagatcagcc cggggcttca
     3721 agtgaaccct tccaggagac cttacggttc ctctctttca ggtgacactt ctccagagaa
     3781 gtcctacggc cgcctaactc ctgttcttcc ctctagtcga attgaagtcc ttcccataga
     3841 cattggtgcc aggggatacg gcagttcagg gtcaccaaag agcggaaagt tcatccttcc
     3901 cccggtggac tatttgcctg agaacagaaa atggtcagaa tttagttaca catacagacc
     3961 tgaggtgcat ggtcagtcat ggactcagga tgctacccgt agccacttaa gcaatttacc
     4021 aaaatacaca gcctttccgc acatacctct tcaggaccac ggccttccag aagtgtctgt
     4081 agacagtcca gtgaggtata agacttcacc agctgtggaa gaggccagtc ctcctgggta
     4141 tccatacgca gggcccacac cttcagcaca tcactacaga aatcgagagg gcctgtctgt
     4201 tcaggaatca gtgttgctat gagcggccgc gactctagag tcgacggatc tttttccctc
     4261 tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag
     4321 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac
     4381 atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac
     4441 atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat
     4501 gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat
     4561 tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca
     4621 tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct
     4681 tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt
     4741 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4801 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4861 tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc
     4921 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4981 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     5041 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     5101 gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5161 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5221 tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5281 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5341 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5401 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5461 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5521 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5581 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5641 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5701 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5761 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5821 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5881 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5941 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6001 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6061 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6121 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6181 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6241 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6301 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6361 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6421 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6481 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6541 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6601 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6661 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6721 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6781 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6841 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6901 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6961 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7021 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7081 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7141 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7201 gcacatttcc ccgaaaa
//
