LOCUS       sequence-448399-        8828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a form of full length SRGAP2 with two point mutations 
            (P339L and F342C) that abrogate EVH1 binding site for Homer1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8828)
  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 8828)
  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_EVH1(P339L F342C)
FEATURES             Location/Qualifiers
     source          1..8828
                     /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..5753)
                     /label=chicken beta-actin promoter
     enhancer        5755..6134
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6165..6269
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6270..6338,6339..7130)
                     /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(6488..6507)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7301..7889
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7790..7809
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8043..8060
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    8128..8262
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8152..8171)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8206..8225
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(8268..8463)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8282..8417
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8522..8543
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8558..8563,8564..8581,8582..8588)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8596..8612
                     /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     8601..8623
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8620..8636
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8620..8636
                     /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 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 cacttcataa 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 cscccccccc
     5641 cccccccgcc cccatcgctg cacaaaataa ttaaaaaata aataaataca aaattggggg
     5701 tggggagggg ggggagatgg ggagagtgaa gcagaacgtg gggctcacct cgaccatggt
     5761 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta
     5821 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg
     5881 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg
     5941 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
     6001 tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg
     6061 gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt aataactagt
     6121 caataatcaa tgtcgaccca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     6181 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6241 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     6301 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6361 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6421 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     6481 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6541 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6601 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6661 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     6721 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6781 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6841 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6901 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6961 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     7021 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     7081 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     7141 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     7201 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     7261 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     7321 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     7381 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7441 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7501 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7561 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7621 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7681 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7741 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7801 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7861 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7921 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7981 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     8041 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     8101 cgcgttggcc gattcattaa tgcagcggat ccagacatga taagatacat tgatgagttt
     8161 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     8221 attgctttat ttgtaaccat tataagctgc aataaacaag ttagcttttt gcaaaagcct
     8281 aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg
     8341 gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc
     8401 ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga
     8461 tgcggatccg ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     8521 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     8581 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     8641 attacgccaa gctggtcgac ggtgcaccgt cgaccagctg gtcgacggtg caggtcgagg
     8701 gatctccata agagaagagg gacagctatg actgggagta gtcaggagag gaggaaaaat
     8761 ctggctagta aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa taacacaaaa
     8821 caaaatat
//
