LOCUS       sequence-329931-        7091 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of F-BAR domain (aa 1-492) of SRGAP3 with a C-terminal 
            EGFP fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7091)
  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 7091)
  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-SRGAP3FBAR-EGFP
FEATURES             Location/Qualifiers
     source          1..7091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        12..391
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        393..670
                     /label=chicken beta-actin promoter
     intron          671..1687
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1611..1633
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1695..1714
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3273..3282
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3279..3281,3282..3284,3285..3998)
                     /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(3279..3281,3282..3284,3285..3995)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3324..3345)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3585..3604)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3932..3953
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4090..4109)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4155..4210
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4209..4228)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4601..4617)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4601..4617)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4614..4636)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4625..4641
                     /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(4649..4655,4656..4673,4674..4679))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4694..4715
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4774..4969
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4820..4955
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4975..5109
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5012..5031)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5066..5085
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5177..5194)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5348..5936)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5428..5447)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6107..6898,6899..6967))
                     /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     6730..6749
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6968..7072)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctgggt cgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
       61 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      121 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      181 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      241 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      301 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      361 catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac
      421 tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt
      481 ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg ccrggsgggg sggggsgggg
      541 sggggggsgg ggsggggsgr ggcggagagg tgcggcggca gccaatcaga gcggcgcgct
      601 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
      661 gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc
      721 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
      781 cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg
      841 gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg
      901 gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg
      961 ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag
     1021 cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt
     1081 gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc
     1141 ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg
     1201 tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc
     1261 gggcggggcg gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg
     1321 gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg
     1381 cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg
     1441 ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat
     1501 gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg
     1561 gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt
     1621 ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt
     1681 cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt
     1741 gctcgagctc aagcttcgaa ttcaagatgt catctcaaac taagttcaag aaagacaaag
     1801 agatcattgc tgaatatgaa gcccaaataa aagagatccg cacgcagctg gtggagcagt
     1861 tcaaatgtct ggagcagcaa tcagagtcgc gactgcagct gcttcaagac ctccaggagt
     1921 ttttccgccg gaaagctgag attgagctcg agtactcccg cagcctggag aagctggctg
     1981 agcgcttctc ctccaaaatc cgcagctccc gggagcacca gttcaagaag gaccagtacc
     2041 tcctctcgcc tgtgaactgt tggtatctgg ttctgcatca gacccggcgg gagagccgag
     2101 accatgccac cctcaatgac atcttcatga acaatgtcat cgtccgcctc tcccagatca
     2161 gtgaggatgt catcagactc ttcaaaaaga gcaaggagat tggcctgcag atgcacgagg
     2221 agctcctgaa ggtgaccaat gagctctaca cagtcatgaa aacctaccac atgtaccatg
     2281 cagagagcat cagtgcggaa agcaagctga aggaggctga gaagcaggag gagaagcagt
     2341 tcaataagtc aggagacctc agcatgaacc tgctccggca cgaggaccgg ccccagcgcc
     2401 gcagctctgt gaagaagatt gagaagatga aggagaagag gcaggccaag tactctgaga
     2461 acaagctgaa atgcacaaag gcccggaatg actacttgct caatctggca gccaccaacg
     2521 cagctataag caaatactac atccatgatg tctctgatct gatcgattgc tgtgatttgg
     2581 gcttccatgc cagcctggcc cgcaccttcc ggacctatct ctcagctgaa tacaacctgg
     2641 agacctctcg ccacgaaggg ctggatgtca ttgagaatgc agtggacaac ctggattccc
     2701 gaagtgacaa gcacacagtc atggacatgt gcaatcaagt cttctgccct ccactcaagt
     2761 tcgagttcca gccccacatg ggggatgagg tctgccaggt cagcgctcag cagcccgtcc
     2821 agacagaact gctcatgcgt tatcaccagc tgcagtccag actggccacc ctcaagatag
     2881 agaatgagga ggttaggaaa accctggatg ccaccatgca gacattacag gacatgctga
     2941 ctgtggagga ctttgatgtc tccgatgcct tccaacacag tcgatcgaca gagtccgtca
     3001 agtcggctgc ctctgagacc tacatgagca agatcaacat tgccaagagg agagccaacc
     3061 agcaggaaac agaaatgttt tattttacaa aatttaaaga gtatgtgaat ggcagtaacc
     3121 tcatcaccaa gctgcaggcc aagcacgatt tactcaagca gaccctgggc gaaggggaaa
     3181 gagcagaatg cggcaccacc agaggaagaa gaaatgctcg aaccaggaac caggattcag
     3241 gaacggtacc gcgggcccgg gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc
     3301 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     3361 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     3421 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     3481 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     3541 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     3601 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     3661 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     3721 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     3781 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     3841 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     3901 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     3961 ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcaat tcactcctca
     4021 ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata
     4081 ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca
     4141 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     4201 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta
     4261 tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct
     4321 ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa
     4381 agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac
     4441 atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct cctgactact
     4501 cccagtcata gctgtccctc ttctcttatg gagatccctc gacctgcacc gtcgaccagc
     4561 tggtcgacgg tgcaccgtcg accagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4621 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4681 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4741 tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt
     4801 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4861 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4921 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg
     4981 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5041 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5101 gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5161 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5221 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5281 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5341 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5401 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5461 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5521 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5581 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5641 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5701 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5761 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5821 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5881 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5941 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6001 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6061 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6121 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6181 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6241 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6301 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6361 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6421 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6481 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6541 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6601 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6661 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6721 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6781 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6841 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6901 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6961 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7021 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7081 cccgaaaagt g
//
