LOCUS       Exported                6116 bp ds-DNA     circular SYN 20-DEC-2021
DEFINITION  WT G3BP1 inserted with GFP tagged on the N-terminus used to stably 
            integrate into cells.
ACCESSION   .
VERSION     .
KEYWORDS    pEGFP-C1-G3BP1-WT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6116)
  AUTHORS   Fischer , J.W., Busa, V.F., Shao, S. Leung, A. K.L.
  TITLE     Structure-mediated RNA decay by UPF1 and G3BP1
  JOURNAL   Molecular Cell, Feb 3, 2020
REFERENCE   2  (bases 1 to 6116)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 20, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6116
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     178..197
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     437..458
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      500..509
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        complement(548..751)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(577..597)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        752..1055
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1230..1818)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1310..1329)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2147..2194
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2219..2238
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2426..3220)
                     /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(2537..2556)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3147..3166
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3255..3612)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3269..3404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3323..3342)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3569..3589
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(3614..3718)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3745..4200
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3832..3851)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4042..4063
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4207..4328
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4218..4237)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4272..4291
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5903..503)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(5903..503)
                     /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(5945..5966)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg
       61 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
      121 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
      181 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
      241 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
      301 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
      361 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
      421 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
      481 agctcctcgc ccttgctcac catggtggcg accggtagcg ctagcggatc tgacggttca
      541 ctaaaccagc tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca
      601 atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac
      661 tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac
      721 gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga
      781 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
      841 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
      901 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
      961 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
     1021 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
     1081 ctaatgaccc cgtaattgat tactattaat aactaatgca tggcggtaat acggttatcc
     1141 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1201 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1261 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1321 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1381 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1441 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1501 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1561 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1621 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     1681 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1741 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1801 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     1861 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     1921 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaacctgag
     1981 gctatggcag ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc acccgaactt
     2041 ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt
     2101 ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac aaacgaccca
     2161 acaccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc ttccggtatt
     2221 gtctccttcc gtgtttcagt tagcctcccc ctagggtggg cgaagaactc cagcatgaga
     2281 tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt
     2341 tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg
     2401 gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga
     2461 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
     2521 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
     2581 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
     2641 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga
     2701 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
     2761 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
     2821 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
     2881 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
     2941 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
     3001 ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc agggcaccgg
     3061 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     3121 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     3181 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
     3241 tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg
     3301 aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca
     3361 tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc
     3421 gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag
     3481 cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg
     3541 cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca cagctggttc
     3601 tttccgcctc aggactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3661 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3721 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3781 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3841 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3901 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3961 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4021 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4081 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     4141 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     4201 acgcgttaag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     4261 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     4321 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     4381 aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcagttat
     4441 ctagatccgg tggatcccgg gcccgcggta ccgtcgactg cagaattccc tgccgtggcg
     4501 caagccccct tcccactcca aatcctggtt tctgcaccat gcctccacgg ggagggcctc
     4561 tcattccacc acccagccca cctcgagggc ctccaggtcc ccgaaggcga ttatctcgtc
     4621 ggtcgccttc cctggcagct cgagtcttct tctcttcgac attcagacgg acctcacctc
     4681 tgaacatgat gggtctgttg ctaaggactt tctgaacagg ctcagaatca tcaaacacaa
     4741 caaaaccaaa attgggtaat ttcccaccac tgttaatgcg caactccacc acgtttccat
     4801 aactttgaaa gaaatcttta agctctgatt tgtccacttc atgaggcagg ttgccaatga
     4861 agagttggtg actgtcaggg tgtctcacca ttcttcgggg ttcaatgtca ccttgctcac
     4921 cagcctcacg gattggtctg ggtccccttt ggggaggaat atttattcgt tgttctcgca
     4981 ctctttgatc ccgctgaggt ctttgtggtg gaatctgaga ttcaggctta gactctggac
     5041 ggggctgtga agctggtact ttaacaacat gaggtggtat cccagtaact ggaacagctc
     5101 cactgggtgg aagattctta ctggtcacag atgcccaaga aaatgtcctc aagtcttcct
     5161 gtactgtctg agctatgtct gcaggtgctg gagaagaact cttctgagca tcctcagggg
     5221 cagtttcttc taatactggc tcaggctttt cctcttggat ttcagataca ggttcttgtt
     5281 ctggttctgg ttcaggatca ggctctggtt cagcaacagg ctcctctaaa tgttcttcca
     5341 tgtcattact gacaactgcc tgatcataga aagttccaga atcatcaggt accacctcag
     5401 gtgtttgctg tctttcttca ggttcctcta cttcttcttc agactcctcc tgaggctcag
     5461 tgacaaaccc accaaagacc tcatcttggt atctgaagat atcattgtga acatagaatt
     5521 tatttgcaac agacccctca ggagcaagga caaacgtttg catgaatctc ctcaaagcct
     5581 ggttgttgtt agagagaagc cccatcacct ggactaccac accatcattt agcgtggcat
     5641 gagcatcaac atggcgaatc ttggtgtggc agttggtgaa gttttgtgac atcactttcc
     5701 tgtggatttc tttctgtccg tagactgcat ctgctggctt tccatttgaa tccaatcccc
     5761 catggacata agaagagttc tttccataaa atctatgcag catgtctggg gcctggttca
     5821 gcagtgtgta atactgtctc acaaattccc gcccgaccag caggggacta ggcttctcca
     5881 tcaccataga tctgagtccg gacttgtaca gctcgtccat gccgagagtg atcccggcgg
     5941 cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcaggg
     6001 cggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg
     6061 tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcgg
//
