LOCUS       sequence-448404-        8829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a form of full length SRGAP2 with one point mutation 
            (W765A) in the SH3 domain abrogating its ability to bind to 
            Gephyrin..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8829)
  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 8829)
  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_SH3(W765A)_GFP
FEATURES             Location/Qualifiers
     source          1..8829
                     /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..5754)
                     /label=chicken beta-actin promoter
     enhancer        5756..6135
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6166..6270
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6271..6339,6340..7131)
                     /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(6489..6508)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7302..7890
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7791..7810
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8044..8061
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    8129..8263
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8153..8172)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8207..8226
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(8269..8464)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8283..8418
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8523..8544
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8559..8564,8565..8582,8583..8589)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8597..8613
                     /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     8602..8624
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8621..8637
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8621..8637
                     /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 ccacgcgtcg
     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 cttcaggaca
     2701 ccagctatgg aatccatgtc atggtcgttc tggtccccag ccagggggtc ctctcctctc
     2761 tcaaaggcat ttttgatgtc attcacttcc acctgggatc ctgacacccg 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 cgcscccccc
     5641 ccccccccgc ccccatcgct gcacaaaata attaaaaaat aaataaatac aaaattgggg
     5701 gtggggaggg gggggagatg gggagagtga agcagaacgt ggggctcacc tcgaccatgg
     5761 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     5821 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     5881 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     5941 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     6001 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     6061 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactag
     6121 tcaataatca atgtcgaccc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6181 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6241 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6301 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6361 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6421 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6481 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6541 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6601 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6661 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6721 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6781 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6841 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6901 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6961 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     7021 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     7081 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     7141 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7201 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7261 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7321 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7381 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7441 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7501 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7561 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7621 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7681 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7741 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7801 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7861 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7921 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     7981 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     8041 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     8101 gcgcgttggc cgattcatta atgcagcgga tccagacatg ataagataca ttgatgagtt
     8161 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     8221 tattgcttta tttgtaacca ttataagctg caataaacaa gttagctttt tgcaaaagcc
     8281 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     8341 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     8401 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     8461 atgcggatcc gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     8521 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     8581 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     8641 gattacgcca agctggtcga cggtgcaccg tcgaccagct ggtcgacggt gcaggtcgag
     8701 ggatctccat aagagaagag ggacagctat gactgggagt agtcaggaga ggaggaaaaa
     8761 tctggctagt aaaacatgta aggaaaattt tagggatgtt aaagaaaaaa ataacacaaa
     8821 acaaaatat
//
