LOCUS       sequence-287409-        6741 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pEGFP-C3 plasmid that expresses Exo70.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6741)
  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 6741)
  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-Exo70
FEATURES             Location/Qualifiers
     source          1..6741
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        54..357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        358..561
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     512..532
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      600..609
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(606..608,609..611,612..1322)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(606..608,609..611,612..1322)
                     /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(651..672)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(912..931)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1259..1280
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3522..3643
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3559..3578)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3613..3632
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3650..4105)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3787..3808)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3999..4018
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4132..4236
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4238..4595
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4238..4429
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4261..4281)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4446..4581
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4630..5424
                     /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             4630..5424
                     /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(4684..4703)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5294..5313
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5612..5631)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5656..5703
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6032..6620
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6521..6540
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
       61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg
      601 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      661 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
      721 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      781 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
      841 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
      901 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
      961 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1021 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1081 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1141 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1201 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1261 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1321 agtactcaga tctcgagatg attcccccac aggaggcatc cgctcgacgg cgggagattg
     1381 aggacaagct gaagcaggag gaggagactc tgtccttcat ccgagacagc ctggagaaga
     1441 gcgaccagct cactaagaac atggtgtcta tcttatcatc ctttgagagc cgccttatga
     1501 agctggagaa ctccatcatc cctgtgcaca agcagacgga gaatctgcag cggctgcagg
     1561 agaatgttga gaagacgctg tcctgcctgg accatgtcat cagctactac catgtggcca
     1621 gtgacactga gaagatcatc agagagggcc ccacaggtag gctggaagag tacctgggaa
     1681 gcatggccaa gattcagaag gcagtggagt atttccagga caacagccca gacagcccgg
     1741 aactcaacaa agtgaaactg ctctttgagc gcgggaagga ggccctggag tccgaatttc
     1801 gcagcctgat gacgcggcac agtaaggtcg tctcgcccgt gctcatcttg gatctgatca
     1861 gtggtgacga tgatctggag gcccaggagg acgtgaccct ggagcacctg cccgagagcg
     1921 tgctccagga tgtcattcgc atctcccgct ggctggtgga atatggccgc aaccaagatt
     1981 tcatgaacgt ctactaccag atacgctcca gccagctgga ccgctccatc aaaggactga
     2041 aggagcattt ccataagagc agttcttcct ctggggttcc ctactcccct gctatcccca
     2101 acaagaggaa agacacacct accaagaagc cagtcaagcg gccagggacg atccgtaagg
     2161 ctcagaacct tctgaaacag tattcccagc atggtctaga tgggaaaaag gggggctcta
     2221 acctcattcc tctggaaggg agagatgaca tgctggacgt ggagaccgat gcctacatcc
     2281 actgcgtcag tgccttcgtc aagctggcgc agagcgagta ccagctgctg gccgacatca
     2341 tccccgagca ccaccagaag aagaccttcg actccctgat acaggatgcc ctggatgggc
     2401 tgatgcttga aggggagaac atcgtgtctg ctgcccggaa ggccattgtg cgacacgact
     2461 tctccacggt gctcaccgtc ttccccatcc tgcgacacct caagcagacc aagcctgagt
     2521 ttgaccaggt gctccagggc acggctgcca gcacaaagaa caagctgcct ggcctcatca
     2581 catccatgga gaccatcggt gccaaagcgc tggaggactt cgcagacaac atcaagaatg
     2641 acccggacaa ggagtacaac atgccgaagg acggcaccgt acacgagctc accagcaatg
     2701 ccatcctctt cctgcagcag cttttggact tccaggagac ggcaggcgcc atgctggcct
     2761 cccaagagac cagctcttcg gccaccagct acagctctga gttcagcaag cggctgctaa
     2821 gcacctatat ctgtaaagtg ctgggcaacc tgcagttgaa cttgctgagc aagtccaagg
     2881 tgtacgagga cccagctctg agcgccatct tcctgcacaa caactacaat tacatcctca
     2941 agtccctgga gaagtctgaa ctgatccagc tggtggcagt gacacagaag actgctgagc
     3001 gctcctaccg ggagcacatt gagcagcaga tccagaccta ccagcgcagc tggttaaagg
     3061 tgactgatta catcgcagag aagaatctac ctgtgttcca gccgggagtc aagctccggg
     3121 acaaggagcg gcagattatc aaggagcgtt ttaagggctt caatgatggc ctcgaagaac
     3181 tgtgcaaaat ccagaaggcc tgggctattc cagacacaga gcagagggac aggattcgcc
     3241 aggcccagaa gaccattgtc aaggagacct acggggcctt tctacagaag tttggcagcg
     3301 tgcccttcac caagaacccg gagaagtaca tcaagtacgg ggtggagcag gtgggcgaca
     3361 tgatcgatcg ccttttcgac acctctgcct gaggatccac cggatctaga taactgatca
     3421 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     3481 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     3541 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3601 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc
     3661 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3721 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3781 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3841 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3901 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3961 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4021 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4081 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4141 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4201 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     4261 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4321 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4381 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4441 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4501 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     4561 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     4621 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4681 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4741 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4801 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4861 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4921 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4981 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5041 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5101 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     5161 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5221 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5281 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5341 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5401 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5461 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5521 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5581 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     5641 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     5701 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     5761 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     5821 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     5881 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5941 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6001 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6061 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6121 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6181 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6241 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6301 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6361 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6421 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6481 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6541 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6601 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6661 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6721 ttaccgccat gcattagtta t
//
