LOCUS       sequence-470046-        6516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing fluorescent Grx1-roGFP2 fusion protein in mammalian cells
            with localization in stress granules..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6516)
  TITLE     Rivera-Fuentes plasmids March 2025
REFERENCE   2  (bases 1 to 6516)
  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"
COMMENT     Sequence Label: G3BP1-Grx1-roGFP2
FEATURES             Location/Qualifiers
     source          1..6516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     140..159
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        414..717
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        718..921
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     872..892
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1004..1013
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2810..2812,2813..3526)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2852..2873)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3113..3132)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3460..3481
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3657..3778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3694..3713)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3748..3767
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3785..4240)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3922..3943)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4134..4153
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4267..4371
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4373..4730
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4373..4564
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4396..4416)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4581..4716
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4765..5559
                     /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             4765..5559
                     /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(4819..4838)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5429..5448
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5747..5766)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5791..5838
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(6167..6516,1..239)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
       61 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      121 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      181 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
      241 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      301 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
      361 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      421 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      481 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      541 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      601 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      661 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      721 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      781 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      841 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      901 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact
      961 cagatctcga gctcaagctt cgaattctgc agtcgacggt accgccacca tggtgatgga
     1021 gaagcctagt cccctgctgg tcgggcggga atttgtgaga cagtattaca cactgctgaa
     1081 ccaggcccca gacatgctgc atagatttta tggaaagaac tcttcttatg tccatggggg
     1141 attggattca aatggaaagc cagcagatgc agtctacgga cagaaagaaa tccacaggaa
     1201 agtgatgtca caaaacttca ccaactgcca caccaagatt cgccatgttg atgctcatgc
     1261 cacgctaaat gatggtgtgg tagtccaggt gatggggctt ctctctaaca acaaccaggc
     1321 tttgaggaga ttcatgcaaa cgtttgtcct tgctcctgag gggtctgttg caaataaatt
     1381 ctatgttcac aatgatatct tcagatacca agatgaggtc tttggtgggt ttgtcactga
     1441 gcctcaggag gagtctgaag aagaagtaga ggaacctgaa gaaagacagc aaacacctga
     1501 ggtggtacct gatgattctg gaactttcta tgatcaggca gttgtcagta atgacatgga
     1561 agaacattta gaggagcctg ttgctgaacc agagcctgat cctgaaccag aaccagaaca
     1621 agaacctgta tctgaaatcc aagaggaaaa gcctgagcca gtattagaag aaactgcccc
     1681 tgaggatgct cagaagagtt cttctccagc acctgcagac atagctcaga cagtacagga
     1741 agacttgagg acattttctt gggcatctgt gaccagtaag aatcttccac ccagtggagc
     1801 tgttccagtt actgggatac cacctcatgt tgttaaagta ccagcttcac agccccgtcc
     1861 agagtctaag cctgaatctc agattccacc acaaagacct cagcgggatc aaagagtgcg
     1921 agaacaacga ataaatattc ctccccaaag gggacccaga ccaatccgtg aggctggtga
     1981 gcaaggtgac attgaacccc gaagaatggt gagacaccct gacagtcacc aactcttcat
     2041 tggcaacctg cctcatgaag tggacaaatc agagcttaaa gatttctttc aaagttatgg
     2101 aaacgtggtg gagttgcgca ttaacagtgg tgggaaatta cccaattttg gttttgttgt
     2161 gtttgatgat tctgagcctg ttcagaaagt ccttagcaac aggcccatca tgttcagagg
     2221 tgaggtccgt ctgaatgtcg aagagaagaa gactcgagct gccagggaag gcgaccgacg
     2281 agataatcgc cttcggggac ctggaggccc tcgaggtggg ctgggtggtg gaatgagagg
     2341 ccctccccgt ggaggcatgg tgcagaaacc aggatttgga gtgggaaggg ggcttgcgcc
     2401 acggcagcag tcgaccggca gcatggctca agagtttgtg aactgcaaaa tccagcctgg
     2461 gaaggtggtt gtgttcatca agcccacctg cccgtactgc aggagggccc aagagatcct
     2521 cagtcaattg cccatcaaac aagggcttct ggaatttgtc gatatcacag ccaccaacca
     2581 cactaacgag attcaagatt atttgcaaca gctcacggga gcaagaacgg tgcctcgagt
     2641 ctttattggt aaagattgta taggcggatg cagtgatcta gtctctttgc aacagagtgg
     2701 ggaactgctg acgcggctaa agcagattgg agctctgcag actgaggagg aggccggcga
     2761 gaagatcaag gagctggaga agaaggccag gaagctgccc gaggaattcg tgagcaaggg
     2821 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2881 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2941 gaagttcatc tccaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     3001 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     3061 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     3121 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     3181 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     3241 ctacaactgc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     3301 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     3361 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcacctg
     3421 ctccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     3481 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagaga gctaagcggc
     3541 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     3601 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     3661 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3721 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     3781 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     3841 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     3901 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     3961 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     4021 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     4081 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     4141 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     4201 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttctggga
     4261 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4321 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     4381 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4441 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4501 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4561 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4621 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     4681 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     4741 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4801 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4861 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4921 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     4981 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5041 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5101 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5161 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5221 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5281 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5341 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5401 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5461 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5521 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5581 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5641 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5701 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     5761 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     5821 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     5881 ccccaccgag accccattgg ggtcaatacg cccgcgtttc ttccttttcc ccaccccacc
     5941 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     6001 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     6061 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6121 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6181 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6241 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6301 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6361 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6421 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6481 gctgaacggg gggttcgtgc acacagccca gcttgg
//
