LOCUS       sequence-329933-        8821 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 8821)
  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 8821)
  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..8821
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(42..630)
                     /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(801..1592,1593..1661))
                     /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     1424..1443
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1662..1766)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1797..2176
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          2456..3472
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3396..3418
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3480..3499
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      6788..6797
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6794..6796,6797..6799,6800..7513)
                     /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(6794..6796,6797..6799,6800..7510)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6839..6860)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7100..7119)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7447..7468
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7605..7624)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7670..7725
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7724..7743)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(8116..8132)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8116..8132)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8129..8151)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8140..8156
                     /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(8164..8170,8171..8188,8189..8194))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8209..8230
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8289..8484
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8335..8470
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    8490..8624
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8527..8546)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8581..8600
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8692..8709)
                     /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 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      541 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      601 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      661 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      721 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      781 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
      841 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
      901 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
      961 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1021 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1081 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1141 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1201 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1261 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1321 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1381 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1441 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1501 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1561 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1621 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1681 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1741 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca
     1801 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     1861 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     1921 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     1981 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     2041 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     2101 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     2161 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
     2221 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg
     2281 ggcggggggg gggggggggc gcgcgccrgg gggggggggg gggggsgggg ggsggggsgg
     2341 ggsgrggcgg rgaggkgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
     2401 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt
     2461 cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg
     2521 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
     2581 ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct
     2641 tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg
     2701 tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg
     2761 gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg
     2821 tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt
     2881 gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc
     2941 tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc
     3001 ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc
     3061 gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg
     3121 tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg
     3181 acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc
     3241 tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
     3301 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg
     3361 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
     3421 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
     3481 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattgctcg acgccatgtc
     3541 atctcaaact aagttcaaga aagacaaaga gatcattgct gaatatgaag cccaaataaa
     3601 agagatccgc acgcagctgg tggagcagtt caaatgtctg gagcagcaat cagagtcgcg
     3661 actgcagctg cttcaagacc tccaggagtt tttccgccgg aaagctgaga ttgagctcga
     3721 gtactcccgc agcctggaga agctggctga gcgcttctcc tccaaaatcc gcagctcccg
     3781 ggagcaccag ttcaagaagg accagtacct cctctcgcct gtgaactgtt ggtatctggt
     3841 tctgcatcag acccggcggg agagccgaga ccatgccacc ctcaatgaca tcttcatgaa
     3901 caatgtcatc gtccgcctct cccagatcag tgaggatgtc atcagactct tcaaaaagag
     3961 caaggagatt ggcctgcaga tgcacgagga gctcctgaag gtgaccaatg agctctacac
     4021 agtcatgaaa acctaccaca tgtaccatgc agagagcatc agtgcggaaa gcaagctgaa
     4081 ggaggctgag aagcaggagg agaagcagtt caataagtca ggagacctca gcatgaacct
     4141 gctccggcac gaggaccggc cccagcgccg cagctctgtg aagaagattg agaagatgaa
     4201 ggagaagagg caggccaagt actctgagaa caagctgaaa tgcacaaagg cccggaatga
     4261 ctacttgctc aatctggcag ccaccaacgc agctataagc aaatactaca tccatgatgt
     4321 ctctgatctg atcgattgct gtgatttggg cttccatgcc agcctggccc gcaccttccg
     4381 gacctatctc tcagctgaat acaacctgga gacctctcgc cacgaagggc tggatgtcat
     4441 tgagaatgca gtggacaacc tggattcccg aagtgacaag cacacagtca tggacatgtg
     4501 caatcaagtc ttctgccctc cactcaagtt cgagttccag ccccacatgg gggatgaggt
     4561 ctgccaggtc agcgctcagc agcccgtcca gacagaactg ctcatgcgtt atcaccagct
     4621 gcagtccaga ctggccaccc tcaagataga gaatgaggag gttaggaaaa ccctggatgc
     4681 caccatgcag acattacagg acatgctgac tgtggaggac tttgatgtct ccgatgcctt
     4741 ccaacacagt cgatcgacag agtccgtcaa gtcggctgcc tctgagacct acatgagcaa
     4801 gatcaacatt gccaagagga gagccaacca gcaggaaaca gaaatgtttt attttacaaa
     4861 atttaaagag tatgtgaatg gcagtaacct catcaccaag ctgcaggcca agcacgattt
     4921 actcaagcag accctgggcg aaggggaaag agcagaatgc ggcaccacca gaggaagaag
     4981 aaatgctcga accaggaacc aggattcagg acaagctata ccgcttgtag tcgagagctg
     5041 catccgttac atcaatttat atggactcca gcagcagggg atcttcagag tgccaggatc
     5101 tcaggtggaa gtcaatgaca tcaaaaattc ctttgagaga ggtgaagacc cccttgtgga
     5161 cgatcaaaat gaacgagata tcaattcagt cgctggtgtt ttaaaactgt atttccgagg
     5221 actggaaaac ccactctttc ctaaggaaag gtttcaagat ttgatatcta ctattaaact
     5281 ggagaaccca gccgagaggg tgcaccagat ccaacaaatc ctcgtcaccc ttccccgcgt
     5341 ggtcattgtg gtcatgagat acctcttcgc tttcctcaac cacctctccc agtatagcga
     5401 cgagaacatg atggatccct acaacctggc catctgcttc gggcctaccc tcatgcacat
     5461 ccctgatggg caggaccctg tgtcctgcca ggcacacatc aatgaagtca tcaaaaccat
     5521 catcatccat catgaagcca tcttccccag cccccgggag ctagagggac ctgtgtatga
     5581 aaaatgcatg gctggagggg aagaatattg tgacagccca cacagcgagc caggggccat
     5641 cgatgaagtt gaccatgaca atggcactga gcctcatacc agcgatgaag aagtggagca
     5701 gatcgaggct attgccaagt ttgactacat ggggcggtcc ccgcgtgagc tatccttcaa
     5761 gaagggggcc tcgctgctcc tgtaccaccg cgcctcggag gactggtggg agggccggca
     5821 caacggcgtg gatggactca tcccccatca gtacatagtt gtacaggaca tggatgatgc
     5881 cttctccgac agcctgagcc agaaggctga cagcgaggcc agcagtgggc cattgctgga
     5941 tgacaaggcc tcttccaaaa acgacctcca gtcccccacg gagcacatct cggattacgg
     6001 ctttgggggg gtgatgggcc gagtgcggtt acgatctgat ggagcagcca tccccagacg
     6061 ccgaagcggg ggcgacacac acagcccgcc ccggggcctg ggccccagca tagacacacc
     6121 accccgggct gctgcctgcc ccagcagccc ccacaaaatc cccctcaccc gggggaggat
     6181 cgagagccct gagaagcgga ggatggcgac gttcgggagc gctggcagca tcaactaccc
     6241 tgacaagaag gcgctctccg aagggcactc gatgaggtcg acctgcggtt ccaccaggca
     6301 cagcagccta ggggaccaca agtccctgga ggccgaggcc ctggcagaag acatcgagaa
     6361 gaccatgagc acggctctgc acgagttgcg ggaactcgag aggcagaaca cggtcaagca
     6421 ggcgccagat gtggtgctgg acaccctgga gcccctgaag aacccgccag gccccgtcag
     6481 ctcggagccc gccagtcccc ttcacaccat cgtcatccgc gaccccgatg ccgccatgcg
     6541 ccgcagcagc agctcctcca ccgagatgat gaccaccttc aagccagccc tgtccgcccg
     6601 cctggctggc gcccagctcc gcccgccccc catgcggccc gtgcggccgg tggtccagca
     6661 ccggtccagc agcagcagca gctcgggcgt gggcagcccg gccgtgacgc ccaccgagaa
     6721 gatgttcccc aacagctcag cggacaagtc gggcaccatg gtttcgcggg cccgggatcc
     6781 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     6841 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     6901 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     6961 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     7021 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     7081 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     7141 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     7201 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     7261 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     7321 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     7381 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     7441 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     7501 gctgtacaag taaagcggcc gcaattcact cctcaggtgc aggctgccta tcagaaggtg
     7561 gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc
     7621 aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa
     7681 tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat
     7741 gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat
     7801 gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa
     7861 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
     7921 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
     7981 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
     8041 ttatggagat ccctcgacct gcaccgtcga ccagctyssy skamyyryrs yswyrrcyrg
     8101 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     8161 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     8221 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     8281 gcggatccgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     8341 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     8401 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     8461 tttggaggcc taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca
     8521 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     8581 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat
     8641 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     8701 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     8761 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     8821 g
//
