LOCUS       sequence-334866-        7118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of F-BAR domain (aa1-516) of SRGAP1 with a C-terminal 
            EGFP fusion..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7118)
  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 7118)
  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-SRGAP1FBAR(516a.a.)-EGFP
FEATURES             Location/Qualifiers
     source          1..7118
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(48..636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(128..147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(807..1598,1599..1667))
                     /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     1430..1449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1668..1772)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1803..2182
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          2445..3461
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3385..3407
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3469..3488
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      5091..5100
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5097..5099,5100..5102,5103..5816)
                     /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(5097..5099,5100..5102,5103..5813)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5142..5163)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5403..5422)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5750..5771
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5908..5927)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5973..6028
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6027..6046)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6419..6435)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6419..6435)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6432..6454)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6443..6459
                     /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(6467..6473,6474..6491,6492..6497))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6512..6533
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6592..6787
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6638..6773
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6793..6927
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6830..6849)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6884..6903
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6995..7012)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
       61 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      121 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      181 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      241 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      301 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      361 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
      421 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      481 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      541 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
      601 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
      661 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
      721 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
      781 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
      841 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
      901 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
      961 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1021 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1081 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1141 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     1201 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     1261 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     1321 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     1381 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     1441 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     1501 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     1561 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     1621 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     1681 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     1741 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctggg
     1801 tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     1861 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     1921 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     1981 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     2041 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     2101 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     2161 attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat
     2221 ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc
     2281 gatgggggcg gggggggggg gggggggggc ggggcgaggg gcggggcggg gcgaggcgga
     2341 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
     2401 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct
     2461 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
     2521 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
     2581 gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc
     2641 gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg
     2701 ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg
     2761 ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt
     2821 gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag
     2881 cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct ccccgagttg
     2941 ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg
     3001 tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg
     3061 gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg
     3121 cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt
     3181 cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct agcgggcgcg
     3241 gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc
     3301 cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg gacggctgcc
     3361 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag
     3421 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     3481 ttattgtgct gtctcatcat tttggcaaag aattgctcga gataatgtcc accccgagcc
     3541 gattcaagaa ggacaaagag atcatagccg agtatgaaag tcaagtcaaa gaaattcgag
     3601 ctcaactggt agaacaacaa aaatgcctgg agcagcaaac ggagatgcga gttcagcttc
     3661 tccaggatct gcaagatttc ttccgaaaaa aagctgaaat tgagacggaa tattcccgga
     3721 atctagagaa gttagcagaa aggttcatgg caaaaacaag aagcactaag gatcatcaac
     3781 aatacaagaa agaccagaac ctgttgtctc cagtgaactg ctggtatttg ctcctgaacc
     3841 aagtaaggag agaaagcaaa gaccatgcaa ccttgagtga catctatctg aacaatgtga
     3901 ttatgcggtt catgcagata agtgaggatt ctaccaggat gtttaaaaag agcaaagaga
     3961 ttgcattcca acttcatgag gatttaatga aggttcttaa tgagctttat acggtgatga
     4021 aaacatacca tatgtatcat gcagagagca tcagtgcaga gagcaagctg aaagaggccg
     4081 aaaaacaaga ggaaaagcaa attgggagat ctggtgatcc agtcttccat attcgactag
     4141 aggagagaca tcaacggcga agctctgtga agaaaattga aaaaatgaaa gaaaaaagac
     4201 aagcaaaata ttcagaaaat aagctaaaat caattaaggc acggaacgaa tatctcctaa
     4261 cacttgaagc caccaatgcc tcagttttca agtactatat tcatgatctt tctgatttaa
     4321 ttgattgctg tgatcttggc taccatgcaa gtctgaacag agccctaaga acatatctgt
     4381 ctgcggagta caaccttgaa acctccagac atgagggctt agacattatt gagaatgcag
     4441 ttgataattt agagcccagg agcgataagc agagattcat ggagatgtac cctgctgcgt
     4501 tctgtccacc aatgaagttt gagtttcagt ctcacatggg tgatgaggtg tgccaggtca
     4561 gtgcccagca gccagtccag gcagagctca tgctcaggta ccaacagttg cagtcccgcc
     4621 ttgccacgct caaaatcgag aatgaagagg ttaagaaaac gactgaagcc accttgcaga
     4681 cgatacaaga tatggtcacc atcgaggact atgatgtttc tgaatgcttc cagcacagtc
     4741 gttccacaga atctgtgaag tccactgtct ctgaaaccta cctgagtaaa cccagcatcg
     4801 ccaagagaag agccaaccag caggaaactg aacagttcta cttcatgaaa ctcagagaat
     4861 atttggaagg cagtaatctc atcacaaaac ttcaagccaa acatgacttg ctgcagagga
     4921 ccctgggaga aggtcataga gctgaatata tgactacaag gcctccaaat gttcccccta
     4981 agccccagaa acacaggaag tccaggcccc gctcacagta taatactaag ttgtttaatg
     5041 gggatttgga aacattcgtc aaggactcag gagcccggga tccaccggtc gccaccatgg
     5101 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     5161 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     5221 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     5281 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     5341 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     5401 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     5461 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     5521 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     5581 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     5641 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     5701 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     5761 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
     5821 gccgcaattc actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa
     5881 tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat
     5941 catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat
     6001 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     6061 aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc
     6121 catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca
     6181 ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt
     6241 gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt
     6301 cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga gatccctcga
     6361 cctgcaccgt cgaccagctg gtcgacggtg caccgtcgac cagcttggcg taatcatggt
     6421 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6481 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6541 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa
     6601 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     6661 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     6721 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     6781 ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6841 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6901 atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa cgcgcgggga
     6961 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     7021 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     7081 aatcagggga taacgcagga aagaacatgt gagcaaaa
//
