LOCUS       sequence-469940-        6975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing fluorescent Grx1-roGFP2 fusion protein in mammalian cells
            with localization in processing bodies..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6975)
  TITLE     Rivera-Fuentes plasmids March 2025
REFERENCE   2  (bases 1 to 6975)
  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: Grx1-roGFP2-Dcp1b
FEATURES             Location/Qualifiers
     source          1..6975
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        97..400
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        401..604
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     555..575
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      687..696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1080..1082,1083..1793)
                     /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(1122..1143)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1383..1402)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1730..1751
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3799..3920
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3836..3855)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3890..3909
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3927..4382)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4064..4085)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4276..4295
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4409..4513
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4515..4872
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4515..4706
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4538..4558)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4723..4858
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4907..5701
                     /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             4907..5701
                     /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(4961..4980)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5571..5590
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5889..5908)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5933..5980
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6309..6897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6798..6817
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
       61 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      121 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      181 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      241 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      301 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      361 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      421 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      481 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      541 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      601 agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct cgagctcaag
      661 cttcgaattc tgcagtcgac ggtaccgcca ccatggctca agagtttgtg aactgcaaaa
      721 tccagcctgg gaaggtggtt gtgttcatca agcccacctg cccgtactgc aggagggccc
      781 aagagatcct cagtcaattg cccatcaaac aagggcttct ggaatttgtc gatatcacag
      841 ccaccaacca cactaacgag attcaagatt atttgcaaca gctcacggga gcaagaacgg
      901 tgcctcgagt ctttattggt aaagattgta taggcggatg cagtgatcta gtctctttgc
      961 aacagagtgg ggaactgctg acgcggctaa agcagattgg agctctgcag actgaggagg
     1021 aggccggcga gaagatcaag gagctggaga agaaggccag gaagctgccc gaggaattcg
     1081 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1141 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1201 agctgaccct gaagttcatc tccaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1261 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     1321 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1381 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1441 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1501 tggagtacaa ctacaactgc cacaacgtct atatcatggc cgacaagcag aagaacggca
     1561 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     1621 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1681 tgagcacctg ctccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1741 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtccggac
     1801 tcagatctcg agctgcagcc gtggcggcag gcggcctggt gggaaagggg cgcgacatca
     1861 gcctagcggc cctgcagcgc cacgacccct atatcaaccg catcgtggac gtggccagcc
     1921 aggtggctct gtacaccttc ggccatcggg ccaacgagtg ggagaaaact gatgtggaag
     1981 gaaccttatt tgtttataca aggtctgctt ctccaaagca tggattcacc attatgaata
     2041 ggctgagcat ggaaaatagg acagaaccta ttactaaaga cttggatttc caactccagg
     2101 accctttcct tctctacaga aatgccagat tgtccatcta tggaatttgg ttttatgata
     2161 aggaagaatg ccaaagaatt gcagagctta tgaaaaacct aactcagtat gaacagttga
     2221 aagcccatca gggaactgga gcaggaattt caccagtgat cctcaattca ggagagggca
     2281 aagaagtaga cattttacga atgctcatca aggccaaaga cgaatacaca aagtgtaaaa
     2341 cctgttctga gccaaaaaag ataaccagtt cctctgccat ctatgacaat ccaaatctca
     2401 tcaaaccaat tccagtgaaa cccagtgaaa accagcaaca gcgtatacct cagcccaacc
     2461 agaccttaga ccctgaaccc caacacttat ccttgacagc tctgtttggg aagcaggaca
     2521 aagctacatg tcaggaaact gtggagcctc cgcagactct ccaccagcag cagcagcagc
     2581 agcagcagca gcagcaagag aagcttccaa ttaggcaggg ggttgtacgc tccctgtcct
     2641 atgaggaacc cagaagacac tcacccccca ttgagaagca gctctgtcca gccattcaga
     2701 aactcatggt caggagcgca gacctccacc cattgtcaga gctgcctgaa aaccggcctt
     2761 gtgaaaatgg cagtacccat tctgcgggag aattttttac aggacctgtc cagccagggt
     2821 ctcctcacaa cattggaact tctcgtggtg tacaaaatgc ttccagaact cagaacctgt
     2881 tcgagaaact tcagagtacc ccaggggcag caaacaagtg tgaccctagt acaccagcac
     2941 ctgccagctc agctgccctg aaccgcagca gagctcccac ttctgtcacc cctgtggctc
     3001 caggaaaggg tctggctcag ccaccacagg cctatttcaa tggctccctt ccacctcaga
     3061 cagtaggaca tcaggctcat ggaagagaac agtccacact cccaagacaa acactcgcca
     3121 tctctggtag tcagactggc agctctggag tgatctcccc tcaagagtta ctgaagaagc
     3181 ttcagattgt acagcaggag cagcagctgc atgcctctaa ccggccagcc ttggccgcta
     3241 agtttcctgt gctcgctcag agctctggaa cagggaaacc cttggaatcc tggatcaaca
     3301 agacacccaa cacagaacag cagactcctc ttttccaggt catctcacct cagcgcatcc
     3361 ctgctacagc agctccgtct ctgcttatgt cccccatggt gttcgcacaa cccacctccg
     3421 tcccgccaaa ggaaagggag agcggcctct tgcctgtggg aggccaggag ccacctgttg
     3481 ctgccaccag cctcctcctg cccatacaga gcccggagcc ctccgtgatc accagcagcc
     3541 cactcaccaa gctccagctc caggaggcac tgctgtacct cattcagaat gatgacaact
     3601 tcttaaatat aatctatgaa gcctatctct tcagcatgac tcaagcagcc atgaaaaaga
     3661 ctatgtgaga gagctaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     3721 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     3781 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3841 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3901 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     3961 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     4021 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     4081 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     4141 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     4201 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     4261 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     4321 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     4381 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4441 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4501 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     4561 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4621 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4681 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4741 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4801 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4861 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4921 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4981 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     5041 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     5101 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     5161 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     5221 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     5281 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     5341 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     5401 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     5461 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5521 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5581 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5641 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5701 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5761 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5821 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     5881 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     5941 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     6001 cagggctggc actctgtcga taccccaccg agaccccatt ggggtcaata cgcccgcgtt
     6061 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     6121 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     6181 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6241 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6301 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6361 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6421 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6481 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6541 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6601 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6661 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6721 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6781 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6841 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6901 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6961 tttcctgcgt tatcc
//
