LOCUS       sequence-456471-        6900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pEGFP-C3 plasmid that expresses Exo84.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6900)
  AUTHORS   Martin TD, Chen XW, Kaplan RE, Saltiel AR, Walker CL, Reiner DJ, Der
            CJ
  TITLE     Ral and Rheb GTPase activating proteins integrate mTOR and GTPase 
            signaling in aging, autophagy, and tumor cell invasion.
  JOURNAL   Mol Cell. 2014 Jan 23;53(2):209-20. doi: 
            10.1016/j.molcel.2013.12.004. Epub 2014 Jan 2.
  PUBMED    24389102
REFERENCE   2  (bases 1 to 6900)
  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"; doi: 
            "10.1016/j.molcel.2013.12.004"; journalName: "Mol Cell"; date: 
            "2014-01-23- 23"; volume: "53"; issue: "2"; pages: "209-20"
COMMENT     Sequence Label: pEGFP-C3-Exo84
FEATURES             Location/Qualifiers
     source          1..6900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3688..3809
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3725..3744)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3779..3798
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3816..4271)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3953..3974)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4165..4184
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4298..4402
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4404..4761
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4404..4595
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4427..4447)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4612..4747
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4796..5590
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4796..5590
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4850..4869)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5460..5479
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5778..5797)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5822..5869
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6198..6786
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6687..6706
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt actcagatct cgagctcaag cttcgaatta tggcgatggc gatgtcggac
     1381 agtggggcga gccgcctgcg tcggcagctg gagtcagggg gttttgaggc gcggctgtac
     1441 gtgaagcagc tctcgcagca gtcggatggg gaccgggacc tccaggagca ccggcagcgc
     1501 atccaggcgc tggcggagga gacggcgcag aacctgaagc gcaacgtcta ccagaactac
     1561 cggcagttca tagagacggc ccgcgagatc tcctacctgg agagcgagat gtaccagctc
     1621 agccatttgc tgaccgagca gaaaagcagc ctggagagca tcccgcttac gttgctgcct
     1681 gccgctgctg ccgccggagc cgccgccgcc tctggagggg aggagggagt cggtggggcg
     1741 gggggccgag accacctccg aggccaggcc ggctttttct ccacccccgg gggtgcctcc
     1801 cgcgacggct ccggtccagg cgaggaagga aagcagcgca ctctcaccac cctgcttgag
     1861 aaggtggaag gctgcaggca tctgctggag acgccgggac agtacttggt gtacaatggg
     1921 gacctagtgg aatacgatgc ggaccacatg gcccaactgc agcgggtgca cggctttctc
     1981 atgaacgatt gcttgttggt ggctacctgg ctgcctcagc ggcgtgggat gtatcgctac
     2041 aacgctctct attccctaga tggtttggcc gtagtcaatg tcaaggacaa cccgcccatg
     2101 aaggacatgt tcaagctgct tatgttcccc gagagccgta ttttccaggc cgaaaatgct
     2161 aaaatcaaac gagagtggct ggaagtgctg gaggacacca agagggccct cagtgagaaa
     2221 aggcgaaggg agcaggagga ggcagcggcc cctcgagggc caccccaagt gacttccaag
     2281 gccactaacc catttgagga tgacgaagaa gaagaaccag ctgttcctga ggtagaggaa
     2341 gagaaggtgg acctctccat ggaatggatc caggagttac ctgaagacct ggatgtctgc
     2401 attgcgcaga gagactttga aggggcggtt gacctgctgg ataaattgaa ccattacctg
     2461 gaagataaac ctagcccacc tcctgtaaaa gaactaaggg ccaaagtgga ggagcgagtt
     2521 cgacagctca ctgaggtgct agttttcgaa ctctccccag atcgttccct gagaggtggt
     2581 ccgaaggcta ctcgcagagc agtttcgcaa ctgatccggc tgggccagtg cacgaaggcc
     2641 tgtgagctat ttttgagaaa cagggcagcc gctgttcata ctgcaattcg tcagcttcgc
     2701 atcgaaggtg ccactttact ctatattcat aagctgtgcc atgtcttctt taccagcctt
     2761 ctcgagactg caagagaatt tgagatcgat tttgcaggca ctgacagcgg ctgctactct
     2821 gcctttgtgg tctgggcaag atcagccatg ggcatgttcg tggatgcttt tagcaagcag
     2881 gtgtttgata gtaaggagag cctctctaca gcagctgagt gtgtaaaagt ggctaaggag
     2941 cattgccagc aactgggtga tatcggactg gatctcacct tcatcatcca tgcccttctg
     3001 gtgaaagaca tccaaggggc cttgcacagt tacaaagaaa tcatcattga agccactaaa
     3061 catcgcaact ctgaagagat gtggaggagg atgaacttga tgacgccaga agccctgggt
     3121 aagctcaaag aagagatgaa aagttgtggg gtaagtaact ttgagcagta cacaggggat
     3181 gactgctggg tgaacctaag ttacacagtg gttgctttca ccaaacagac catgggcttc
     3241 ttggaagagg ccctgaagct gtatttccca gagctgcaca tggtactttt ggagagcctg
     3301 gtggaaatca ttttggttgc tgttcagcat gtggattata gtcttcgatg tgagcaggat
     3361 ccagagaaga aagcttttat cagacagaat gcatcctttt tatatgaaac agtcctccct
     3421 gtggtggaga aaaggtttga agaaggtgtg gggaaacctg ccaagcaact ccaagatctg
     3481 aggaatgcat ctagacttat tcgtgtgaat cctgaaagta caacatcagt ggtctaagtc
     3541 gacggtaccg cgggcccggg atccaccgga tctagataac tgatcataat cagccatacc
     3601 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     3661 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     3721 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3781 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
     3841 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     3901 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     3961 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     4021 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     4081 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     4141 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     4201 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     4261 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4321 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4381 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     4441 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4501 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4561 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4621 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4681 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4741 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     4801 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4861 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4921 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     4981 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     5041 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     5101 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     5161 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     5221 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     5281 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     5341 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     5401 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5461 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5521 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5581 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     5641 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5701 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5761 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     5821 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     5881 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     5941 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     6001 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     6061 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6121 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6181 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6241 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6301 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6361 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6421 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6481 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6541 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6601 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6661 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6721 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6781 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6841 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
