LOCUS       sequence-326692-        7130 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 7130)
  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 7130)
  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..7130
                     /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      5097..5106
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5103..5105,5106..5108,5109..5822)
                     /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(5103..5105,5106..5108,5109..5819)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5148..5169)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5409..5428)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5756..5777
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5914..5933)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5979..6034
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6033..6052)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6425..6441)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6425..6441)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6438..6460)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6449..6465
                     /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(6473..6479,6480..6497,6498..6503))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6518..6539
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6598..6793
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6644..6779
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6799..6933
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6836..6855)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6890..6909
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7001..7018)
                     /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 gctcgagata atgtccaccc
     3541 cgagccgatt caagaaggac aaagagatca tagccgagta tgaaagtcaa gtcaaagaaa
     3601 ttcgagctca actggtagaa caacaaaaat gcctggagca gcaaacggag atgcgagttc
     3661 agcttctcca ggatctgcaa gatttcttcc gaaaaaaagc tgaaattgag acggaatatt
     3721 cccggaatct agagaagtta gcagaaaggt tcatggcaaa aacaagaagc actaaggatc
     3781 atcaacaata caagaaagac cagaacctgt tgtctccagt gaactgctgg tatttgctcc
     3841 tgaaccaagt aaggagagaa agcaaagacc atgcaacctt gagtgacatc tatctgaaca
     3901 atgtgattat gcggttcatg cagataagtg aggattctac caggatgttt aaaaagagca
     3961 aagagattgc attccaactt catgaggatt taatgaaggt tcttaatgag ctttatacgg
     4021 tgatgaaaac ataccatatg tatcatgcag agagcatcag tgcagagagc aagctgaaag
     4081 aggccgaaaa acaagaggaa aagcaaattg ggagatctgg tgatccagtc ttccatattc
     4141 gactagagga gagacatcaa cggcgaagct ctgtgaagaa aattgaaaaa atgaaagaaa
     4201 aaagacaagc aaaatattca gaaaataagc taaaatcaat taaggcacgg aacgaatatc
     4261 tcctaacact tgaagccacc aatgcctcag ttttcaagta ctatattcat gatctttctg
     4321 atttaattga ttgctgtgat cttggctacc atgcaagtct gaacagagcc ctaagaacat
     4381 atctgtctgc ggagtacaac cttgaaacct ccagacatga gggcttagac attattgaga
     4441 atgcagttga taatttagag cccaggagcg ataagcagag attcatggag atgtaccctg
     4501 ctgcgttctg tccaccaatg aagtttgagt ttcagtctca catgggtgat gaggtgtgcc
     4561 aggtcagtgc ccagcagcca gtccaggcag agctcatgct caggtaccaa cagttgcagt
     4621 cccgccttgc cacgctcaaa atcgagaatg aagaggttaa gaaaacgact gaagccacct
     4681 tgcagacgat acaagatatg gtcaccatcg aggactatga tgtttctgaa tgcttccagc
     4741 acagtcgttc cacagaatct gtgaagtcca ctgtctctga aacctacctg agtaaaccca
     4801 gcatcgccaa gagaagagcc aaccagcagg aaactgaaca gttctacttc atgaaactca
     4861 gagaatattt ggaaggcagt aatctcatca caaaacttca agccaaacat gacttgctgc
     4921 agaggaccct gggagaaggt catagagctg aatatatgac tacaaggcct ccaaatgttc
     4981 cccctaagcc ccagaaacac aggaagtcca ggccccgctc acagtataat actaagttgt
     5041 ttaatgggga tttggaaaca ttcgtcaagg actcaggagc ccgggatcca ccggtcgcca
     5101 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     5161 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     5221 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     5281 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     5341 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     5401 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     5461 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     5521 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     5581 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     5641 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     5701 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     5761 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     5821 aaagcggccg caattcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     5881 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     5941 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     6001 tgcaatagtg tgttggaatt ttttgtgtct atcactcgga aggacatatg ggagggcaaa
     6061 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     6121 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     6181 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     6241 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     6301 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc
     6361 cctcgacctg caccgtcgac cagctggtcg acggtgcacc gtcgaccagc ttggcgtaat
     6421 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     6481 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     6541 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatccgca
     6601 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     6661 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     6721 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     6781 aggcttttgc aaaaagctaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     6841 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6901 aaactcatca atgtatctta tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg
     6961 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     7021 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     7081 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
//
