LOCUS       sequence-326690-        8816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of full length SRGAP3 with a C-terminal EGFP fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8816)
  AUTHORS   Coutinho-Budd J, Ghukasyan V, Zylka MJ, Polleux F
  TITLE     The F-BAR domains from srGAP1, srGAP2 and srGAP3 regulate membrane 
            deformation differently.
  JOURNAL   J Cell Sci. 2012 Jul 15;125(Pt 14):3390-401. doi: 
            10.1242/jcs.098962. Epub 2012 Mar 30.
  PUBMED    22467852
REFERENCE   2  (bases 1 to 8816)
  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"; journalName: "J Cell 
            Sci."; date: "2012-07-15"; volume: "125(Pt 14):3390-401. doi"; 
            pages: " 10.1242/jcs.098962. Epub 2012 Mar 30"
COMMENT     Sequence Label: pCAG-SRGAP3-EGFP
FEATURES             Location/Qualifiers
     source          1..8816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(42..629)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(122..141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(800..1591,1592..1660))
                     /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     1423..1442
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1661..1765)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1796..2175
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2177..2454
                     /label=chicken beta-actin promoter
     intron          2455..3467
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3391..3413
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3475..3494
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      6783..6792
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6789..6791,6792..6794,6795..7508)
                     /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             join(6789..6791,6792..6794,6795..7505)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6834..6855)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7095..7114)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7442..7463
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7600..7619)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7665..7720
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7719..7738)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(8111..8127)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8111..8127)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8124..8146)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8135..8151
                     /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(8159..8165,8166..8183,8184..8189))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8204..8225
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8284..8479
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8330..8465
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    8485..8619
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8522..8541)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8576..8595
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8687..8704)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
       61 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      121 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      181 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      241 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      301 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      361 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      421 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      481 agacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
      541 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
      601 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
      661 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
      721 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
      781 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
      841 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
      901 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
      961 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     1021 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     1081 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     1141 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     1201 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     1261 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     1321 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     1381 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     1441 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     1501 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     1561 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     1621 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     1681 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     1741 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gggtcgacat
     1801 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     1861 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     1921 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     1981 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     2041 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     2101 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     2161 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc
     2221 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
     2281 gcgggggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg
     2341 gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
     2401 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
     2461 gctgcgttgc cttcgccccg tgccccgctc cgcgccgcct cgcgccgccc gccccggctc
     2521 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
     2581 aattagcgct tggtttaatg acggctcgtt tcttttctgt ggctgcgtga aagccttaaa
     2641 gggctccggg agggcccttt gtgcgggggg gagcggctcg gggggtgcgt gcgtgtgtgt
     2701 gtgcgtgggg agcgccgcgt gcggcccgcg ctgcccggcg gctgtgagcg ctgcgggcgc
     2761 ggcgcggggc tttgtgcgct ccgcgtgtgc gcgaggggag cgcggccggg ggcggtgccc
     2821 cgcggtgcgg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg
     2881 gtgagcaggg ggtgtgggcg cggcggtcgg gctgtaaccc ccccctgcac ccccctcccc
     2941 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc
     3001 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc
     3061 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
     3121 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
     3181 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
     3241 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
     3301 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
     3361 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     3421 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     3481 tgctggttat tgtgctgtct catcattttg gcaaagaatt gctcgacgcc atgtcatctc
     3541 aaactaagtt caagaaagac aaagagatca ttgctgaata tgaagcccaa ataaaagaga
     3601 tccgcacgca gctggtggag cagttcaaat gtctggagca gcaatcagag tcgcgactgc
     3661 agctgcttca agacctccag gagtttttcc gccggaaagc tgagattgag ctcgagtact
     3721 cccgcagcct ggagaagctg gctgagcgct tctcctccaa aatccgcagc tcccgggagc
     3781 accagttcaa gaaggaccag tacctcctct cgcctgtgaa ctgttggtat ctggttctgc
     3841 atcagacccg gcgggagagc cgagaccatg ccaccctcaa tgacatcttc atgaacaatg
     3901 tcatcgtccg cctctcccag atcagtgagg atgtcatcag actcttcaaa aagagcaagg
     3961 agattggcct gcagatgcac gaggagctcc tgaaggtgac caatgagctc tacacagtca
     4021 tgaaaaccta ccacatgtac catgcagaga gcatcagtgc ggaaagcaag ctgaaggagg
     4081 ctgagaagca ggaggagaag cagttcaata agtcaggaga cctcagcatg aacctgctcc
     4141 ggcacgagga ccggccccag cgccgcagct ctgtgaagaa gattgagaag atgaaggaga
     4201 agaggcaggc caagtactct gagaacaagc tgaaatgcac aaaggcccgg aatgactact
     4261 tgctcaatct ggcagccacc aacgcagcta taagcaaata ctacatccat gatgtctctg
     4321 atctgatcga ttgctgtgat ttgggcttcc atgccagcct ggcccgcacc ttccggacct
     4381 atctctcagc tgaatacaac ctggagacct ctcgccacga agggctggat gtcattgaga
     4441 atgcagtgga caacctggat tcccgaagtg acaagcacac agtcatggac atgtgcaatc
     4501 aagtcttctg ccctccactc aagttcgagt tccagcccca catgggggat gaggtctgcc
     4561 aggtcagcgc tcagcagccc gtccagacag aactgctcat gcgttatcac cagctgcagt
     4621 ccagactggc caccctcaag atagagaatg aggaggttag gaaaaccctg gatgccacca
     4681 tgcagacatt acaggacatg ctgactgtgg aggactttga tgtctccgat gccttccaac
     4741 acagtcgatc gacagagtcc gtcaagtcgg ctgcctctga gacctacatg agcaagatca
     4801 acattgccaa gaggagagcc aaccagcagg aaacagaaat gttttatttt acaaaattta
     4861 aagagtatgt gaatggcagt aacctcatca ccaagctgca ggccaagcac gatttactca
     4921 agcagaccct gggcgaaggg gaaagagcag aatgcggcac caccagagga agaagaaatg
     4981 ctcgaaccag gaaccaggat tcaggacaag ctataccgct tgtagtcgag agctgcatcc
     5041 gttacatcaa tttatatgga ctccagcagc aggggatctt cagagtgcca ggatctcagg
     5101 tggaagtcaa tgacatcaaa aattcctttg agagaggtga agaccccctt gtggacgatc
     5161 aaaatgaacg agatatcaat tcagtcgctg gtgttttaaa actgtatttc cgaggactgg
     5221 aaaacccact ctttcctaag gaaaggtttc aagatttgat atctactatt aaactggaga
     5281 acccagccga gagggtgcac cagatccaac aaatcctcgt cacccttccc cgcgtggtca
     5341 ttgtggtcat gagatacctc ttcgctttcc tcaaccacct ctcccagtat agcgacgaga
     5401 acatgatgga tccctacaac ctggccatct gcttcgggcc taccctcatg cacatccctg
     5461 atgggcagga ccctgtgtcc tgccaggcac acatcaatga agtcatcaaa accatcatca
     5521 tccatcatga agccatcttc cccagccccc gggagctaga gggacctgtg tatgaaaaat
     5581 gcatggctgg aggggaagaa tattgtgaca gcccacacag cgagccaggg gccatcgatg
     5641 aagttgacca tgacaatggc actgagcctc ataccagcga tgaagaagtg gagcagatcg
     5701 aggctattgc caagtttgac tacatggggc ggtccccgcg tgagctatcc ttcaagaagg
     5761 gggcctcgct gctcctgtac caccgcgcct cggaggactg gtgggagggc cggcacaacg
     5821 gcgtggatgg actcatcccc catcagtaca tagttgtaca ggacatggat gatgccttct
     5881 ccgacagcct gagccagaag gctgacagcg aggccagcag tgggccattg ctggatgaca
     5941 aggcctcttc caaaaacgac ctccagtccc ccacggagca catctcggat tacggctttg
     6001 ggggggtgat gggccgagtg cggttacgat ctgatggagc agccatcccc agacgccgaa
     6061 gcgggggcga cacacacagc ccgccccggg gcctgggccc cagcatagac acaccacccc
     6121 gggctgctgc ctgccccagc agcccccaca aaatccccct cacccggggg aggatcgaga
     6181 gccctgagaa gcggaggatg gcgacgttcg ggagcgctgg cagcatcaac taccctgaca
     6241 agaaggcgct ctccgaaggg cactcgatga ggtcgacctg cggttccacc aggcacagca
     6301 gcctagggga ccacaagtcc ctggaggccg aggccctggc agaagacatc gagaagacca
     6361 tgagcacggc tctgcacgag ttgcgggaac tcgagaggca gaacacggtc aagcaggcgc
     6421 cagatgtggt gctggacacc ctggagcccc tgaagaaccc gccaggcccc gtcagctcgg
     6481 agcccgccag tccccttcac accatcgtca tccgcgaccc cgatgccgcc atgcgccgca
     6541 gcagcagctc ctccaccgag atgatgacca ccttcaagcc agccctgtcc gcccgcctgg
     6601 ctggcgccca gctccgcccg ccccccatgc ggcccgtgcg gccggtggtc cagcaccggt
     6661 ccagcagcag cagcagctcg ggcgtgggca gcccggccgt gacgcccacc gagaagatgt
     6721 tccccaacag ctcagcggac aagtcgggca ccatggtttc gcgggcccgg gatccaccgg
     6781 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     6841 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     6901 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     6961 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     7021 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     7081 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     7141 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     7201 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     7261 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     7321 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     7381 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     7441 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     7501 acaagtaaag cggccgcaat tcactcctca ggtgcaggct gcctatcaga aggtggtggc
     7561 tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa
     7621 ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat
     7681 tttcattgca atagtgtgtt ggaatttttt gtgtctatca ctcggaagga catatgggag
     7741 ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca
     7801 tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc
     7861 ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat
     7921 attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact
     7981 agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg
     8041 gagatccctc gacctgcacc gtcgaccagc tggtcgacgg tgcaccgtcg accagcttgg
     8101 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     8161 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     8221 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga
     8281 tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     8341 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     8401 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     8461 aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa
     8521 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     8581 ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc
     8641 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     8701 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     8761 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccag
//
