LOCUS       sequence-448408-        8830 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses full length SRGAP2 with R527L, K566A mt in the 
            Rac1-specific GAP domain that abrogates the GTPase activating 
            function of SRGAP2 without locking the GAP-domain in a form bound to
            Rac1-GDP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8830)
  AUTHORS   Fossati M, Pizzarelli R, Schmidt ER, Kupferman JV, Stroebel D, 
            Polleux F, Charrier C
  TITLE     SRGAP2 and Its Human-Specific Paralog Co-Regulate the Development of
            Excitatory and Inhibitory Synapses.
  JOURNAL   Neuron. 2016 Jul 20;91(2):356-69. doi: 10.1016/j.neuron.2016.06.013.
            Epub 2016 Jun 30.
  PUBMED    27373832
REFERENCE   2  (bases 1 to 8830)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.neuron.2016.06.013"; journalName: "Neuron"; date: 
            "2016-07-20- 20"; volume: "91"; issue: "2"; pages: "356-69"
COMMENT     Sequence Label: pCAG_SRGAP2A_GAP(R527L_K566A)
FEATURES             Location/Qualifiers
     source          1..8830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     181..200
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    199..254
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     300..319
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(411..1124,1125..1127,1128..1130))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(414..1124,1125..1127,1128..1130))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(456..477)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     805..824
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1064..1085
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1127..1136
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4435..4454)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          4462..5478
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(4516..4538)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(5479..5755)
                     /label=chicken beta-actin promoter
     enhancer        5757..6136
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6167..6271
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6272..6340,6341..7132)
                     /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     complement(6490..6509)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7303..7891
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7792..7811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8045..8062
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    8130..8264
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8154..8173)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8208..8227
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(8270..8465)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8284..8419
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8524..8545
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8560..8565,8566..8583,8584..8590)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8598..8614
                     /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)."
     primer_bind     8603..8625
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8622..8638
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8622..8638
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaaaaaaatc taacctcaag tcaaggcttt tctatggaat aaggaatgga cagcaggggg
       61 ctgtttcata tactgatgac ctctttatag ccaacctttg ttcatggcag ccagcatatg
      121 ggcatatgtt gccaaactct aaaccaaata ctcattctga tgttttaaat gatttgccct
      181 cccatatgtc cttccgagtg agagacacaa aaaattccaa cacactattg caatgaaaat
      241 aaatttcctt tattagccag aagtcagatg ctcaaggggc ttcatgatgt ccccataatt
      301 tttggcagag ggaaaaagat ctcagtggta tttgtgagcc agggcattgg ccacaccagc
      361 caccaccttc tgataggcag cctgcacctg aggagtgaat tgcggccgct ttacttgtac
      421 agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga
      481 tcgcgcttct cgttggggtc tttgctcagg gcggactggg tgctcaggta gtggttgtcg
      541 ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc
      601 acgctgccgt cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc gttcttctgc
      661 ttgtcggcca tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg
      721 atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg
      781 ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg
      841 cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg
      901 tcggggtagc ggctgaagca ctgcacgccg taggtcaggg tggtcacgag ggtgggccag
      961 ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca
     1021 tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg
     1081 accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc
     1141 ggtggatccc gggcccgcgg taccgtcgat cgagcgacag tacaagactt gtcagtagac
     1201 tgtggggaag ttgatgagtt gacaccaggg ctagtggcat ttgttttggg gaagacagtg
     1261 ggcttgggcc gtgtggctgg tggtttgacc ggggcagcca tcttgacaga cttgacaggc
     1321 cggaaggcgc aggcgatgtc cccagcagta ctggcgctgc gctggaaggc gggctcgggg
     1381 tccttgagga gctgggtgtg cagagggctg gagggctctg acgtgggggc caccactggg
     1441 gaggttttga ggggctccaa ggtgtccaga accacgtcag gggtgtgttt gacactgctc
     1501 tgccgttcta gttcccgtag ctcattcagg gccgagttca ttgttgcctc aatatcctga
     1561 gcaatgacct cagggtccat gggccgatgg ttattgaagc tttttgacct tcccgctgtg
     1621 gcagtcttcc ggatctgtgg actatccagc cgattcttca gggatgagtg gcggctgatg
     1681 gagttgaggc tcccgtgccc actgatggaa cacttatctg gcccttcttt ggggaggctc
     1741 tggctcatga tgggctggga ggatgggcgg cagctagccc ccacccctgg ggaggaggag
     1801 tcagtcaggg aactgctcag cccatggctg tcactccgaa aagttttccg gatgctccca
     1861 gattctggac gcttcctttg cttgttgatg tttgcaagat aaatatcggc tacatgaccc
     1921 ccactgggac agctggcccc cgctctggct gtcacctttt cttcaggtgg ctcagaaatg
     1981 acctcaatct cagacttggg gctggacctc tccacgacac cgtcctcggt gtcttggacc
     2041 acgatgtact gatgggggat gagtccgtcg atgccattgt gccggccttc ccaccagtcg
     2101 tcggaagccc gctggtaaag cagcagggat gctcccttct taaaggacag ctctcgggct
     2161 gtccggccca cgtagtcaaa cttggcaatg gcctcgatgg gctcacattc gtcatcgctc
     2221 gtgtggtgct ctgcggtcac atcctgggtt gagtcttcaa ccgaggtagt ctctccatga
     2281 gggctatcac agtaatcctc catgcttcct cctctgctgt agacagggcc ctccagctcc
     2341 ctggggcttg ggaagatgtt ctcatgctgg atgatgatgg ttttgatcag ctcattcacg
     2401 tgggcttggc aggacacctg gtcgtggccc tctggcactg acattagcga gggcccgaag
     2461 cagatggcga ggttgtaggg gtccatcatg ttctcttcac tgaactgtga taaatgattg
     2521 aggaaggcaa agaggtatct catgataatc agagtggttt tgggcaggac taggaggact
     2581 ttccggatgt gcagagctct ctcctgcagg ttgtccattg tgacgcaggc catcaggtca
     2641 tgaaagatgt ccttggggaa gagagggtgt tccagccccc ggaagtaaag cgccaggaca
     2701 ccagctatgg aatccatgtc atggtcgttc tggtccccag ccagggggtc ctctcctctc
     2761 tcaaaggcat ttttgatgtc attcacttcc acctgggatc ctgacaccag gaaaattcct
     2821 tcatgctgta gtccgtgtct gctgataaac cggatacagc tttccaccac cagaggaatt
     2881 gcctggctgg agtcctgttt cctcacagtt gagctgcgcc tggctagact gcaatctgtc
     2941 cgctgacttt ctcccagggt tttctgcaga aggtcatgct tggcttgtaa cttggtgatg
     3001 aggttcctgc cctccaggta ctctttcatt tttgtgaaat aaaactgctc tgtctcttgc
     3061 tggttggctc tcctcttagc aatgctgggc ttgctcatga aggtttcaga gaccgtggac
     3121 ttgacggact ccatggagtt gctgtactgg aagcagtcag acacatcaaa gtcctcgaca
     3181 gtcacaatgt cctggatggt ttgcagggtg gcctccattg tcttctttac ctcttcgttt
     3241 tcaatcttta gagtggataa gcgagactgc agttgttggc atctctgtac cagctcactc
     3301 tggacaggct gctgggcaca gagctgggaa gccatatccc ccatgtgggg ctgaaactca
     3361 aacttcatag gggggcagaa gacgttgttg tacatctcca tgaggcgctg cttgtcactg
     3421 gtggcatcca ggttttctac tgcattctcg atggcatcca gaccctcatg cttcgactgt
     3481 tccaggttta actcagcaga gaggaaggtg cgtagagccc ggttcagact tgcatggtag
     3541 cctaagtcac aacactgatc aataaggtca gataggtcat ggatgtagta cttgaagaca
     3601 gatgcattgg ttgcctccaa agccagcaag tactcattcc gggctttgat ggccttcagc
     3661 ttattctccg tgtacttggc ttgacgcttc tccttcatct tctcaatctt cttcactgag
     3721 ctcctccgga catgtttctc ctcaatgcga acgttggccg tggagtcagg ggagcgtggg
     3781 gtctgccggt cctcctgctt taccgattta ccaatttgct tctcctcctg cttctccgcc
     3841 tcctttagtt tgctctgagc actgatgctg tcggcattgt acatgtgata tgtcttcatc
     3901 accgagtaga gctcgttcag gaccttcatc aaatcatctt ggagctgctg gccgacttct
     3961 ttactctttt taaagagtct tcctgagtcc tcgctgactt gtacaaatcg aggaatgata
     4021 ttattcaggt agatgtcact cagggtggta tggtccctgc tttcccgctt cacctggttt
     4081 aagaggagat tccagcagtt gactggagag agaacattct gatccttctt gaattgctgg
     4141 tccttggtgc tgcgtgtctt ggccaggaag cgttctgcca gcttctccag gttgcgggag
     4201 tagtccatct caatctctgc cttctttcgg aagaagtcct ggaggtcctg caacagttgc
     4261 acccgaagct cacactgctg gtccaggcat ttcatctgct ctgtgagctg agcacggatc
     4321 tctttgacct gagtatcgta ctctgctatg atctccttat cctttttgaa tttggctgga
     4381 gacgtcatga attagcttga gctcgagcaa ttctttgcca aaatgatgag acagcacaat
     4441 aaccagcacg ttgcccagga gctgtaggaa aaagaagaag gcatgaacat ggttagcaga
     4501 ggctctagag ccgccggtca cacgccagaa gccgaacccc gccctgcccc gtcccccccg
     4561 aaggcagccg tccccccgcg gacagccccg aggctggaga gggagaaggg gacggcggcg
     4621 cggcgacgca cgaaggccct ccccgcccat ttccttcctg ccggcgccgc accgcttcgc
     4681 cccgcgcccg ctagaggggg tgcggcggcg cctcccagat ttcggctccg cacagatttg
     4741 ggacaaagga agtccctgcg ccctctcgca cgattaccat aaaaggcaat ggctgcggct
     4801 cgccgcgcct cgacagccgc cggcgctccg ggggccgccg cgcccctccc ccgagccctc
     4861 cccggcccga ggcggccccg ccccgcccgg cacccccacc tgccgccacc ccccgcccgg
     4921 cacggcgagc cccgcgccac gccccgtacg gagccccgca cccgaagccg ggccgtgctc
     4981 agcaactcgg ggaggggggt gcaggggggg ttgcagcccg accgacgcgc ccacaccccc
     5041 tgctcacccc cccacgcaca caccccgcac gcagcctttg ttcccctcgc agcccccccc
     5101 gcaccgcggg gcaccgcccc cggccgcgct cccctcgcgc acactgcgga gcgcacaaag
     5161 ccccgcgccg cgcccgcagc gctcacagcc gccgggcagc gcggagccgc acgcggcgct
     5221 ccccacgcac acacacacgc acgcaccccc cgagccgctc cccccgcaca aagggccctc
     5281 ccggagcccc tcaaggcttt cacgcagcca cagaaaagaa acaagccgtc attaaaccaa
     5341 gcgctaatta cagcccggag gagaagggcc gtcccgcccg ctcacctgtg ggagtaacgc
     5401 ggtcagtcag agccggggcg ggcggcgcga ggcggcggcg gagcggggca cggggcgaag
     5461 gcagcgcgca gcgactcccg cccgccgcgc gcttcgcttt ttatagggcc gccgccgccg
     5521 ccgcctcgcc ataaaaggaa actttcggag cgcgccgctc tgattggctg ccgccgcacc
     5581 tctccgcctc gccccgcccc gcccctcgcc ccgccccgcc ccgcctggcg cgcgcccccc
     5641 cccccccccg cccccatcgc tgcacaaaat aattaaaaaa taaataaata caaaattggg
     5701 ggtggggagg ggggggagat ggggagagtg aagcagaacg tggggctcac ctcgaccatg
     5761 gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg
     5821 tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt
     5881 ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat
     5941 tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc
     6001 aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg
     6061 cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta
     6121 gtcaataatc aatgtcgacc caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6181 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6241 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     6301 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6361 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     6421 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     6481 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6541 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6601 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6661 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6721 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6781 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     6841 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6901 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6961 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     7021 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     7081 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     7141 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     7201 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     7261 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     7321 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     7381 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     7441 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     7501 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7561 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7621 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7681 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7741 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7801 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7861 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7921 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     7981 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     8041 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     8101 cgcgcgttgg ccgattcatt aatgcagcgg atccagacat gataagatac attgatgagt
     8161 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     8221 ctattgcttt atttgtaacc attataagct gcaataaaca agttagcttt ttgcaaaagc
     8281 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct
     8341 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     8401 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     8461 gatgcggatc cgctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     8521 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     8581 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     8641 tgattacgcc aagctggtcg acggtgcacc gtcgaccagc tggtcgacgg tgcaggtcga
     8701 gggatctcca taagagaaga gggacagcta tgactgggag tagtcaggag aggaggaaaa
     8761 atctggctag taaaacatgt aaggaaaatt ttagggatgt taaagaaaaa aataacacaa
     8821 aacaaaatat
//
