LOCUS       sequence-469938-        6852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing fluorescent Grx1-roGFP2 fusion protein in mammalian cells
            with localization in Cajal bodies..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6852)
  TITLE     Rivera-Fuentes plasmids March 2025
REFERENCE   2  (bases 1 to 6852)
  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-Coilin
FEATURES             Location/Qualifiers
     source          1..6852
                     /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    3676..3797
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3713..3732)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3767..3786
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3804..4259)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3941..3962)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4153..4172
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4286..4390
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4392..4749
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4392..4583
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4415..4435)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4600..4735
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4784..5578
                     /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             4784..5578
                     /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(4838..4857)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5448..5467
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5766..5785)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5810..5857
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6186..6774
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6675..6694
                     /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 aagtactcag
     1801 atccaccaag caagatggca gcttccgaga cggttaggct acggcttcaa tttgattacc
     1861 cgccgccagc taccccgcac tgtacggcct tctggcttct ggtcgacttg aacagatgcc
     1921 gagtcgtcac agatctcatt agtctcatcc gccagcgctt cggcttcagt tctggggcct
     1981 tcctaggcct ctacctggag ggggggctct tgccccccgc cgagagcgcg cgccttgtga
     2041 gagacaacga ctgcctcaga gttaaattag aagagagagg agttgctgag aattctgtag
     2101 tcatcagtaa tggtgacatt aatttatctc ttagaaaagc aaagaagcgg gcatttcagt
     2161 tagaggaggg tgaagaaact gaaccagatt gcaaatattc aaagaagcat tggaagagtc
     2221 gagagaacaa taacaataat gagaaggtct tggatctgga accaaaagct gtcacagatc
     2281 agactgtcag caaaaaaaac aagagaaaaa ataaagcaac ctgtggcaca gtgggtgatg
     2341 ataacgaaga ggccaaaaga aaatcaccaa agaaaaagga gaaatgtgaa tataaaaaaa
     2401 aggctaagaa tcccaagtct ccgaaagtac aggcagtgaa agactgggcc aatcagagat
     2461 gtagttctcc aaaaggttct gctagaaaca gccttgttaa agccaaaagg aaaggtagtg
     2521 taagcgtttg ctcaaaagag agtcccagtt cctcctcgga gtctgagtct tgtgatgaat
     2581 ctatcagtga tggtcccagc aaagtcactt tggaggccag aaattcctca gagaaattac
     2641 caactgagtt atcaaaggaa gaaccctcta ccaaaaatac aactgcagac aaactggcta
     2701 taaaacttgg ctttagcctt acccccagca agggcaagac ctctggaaca acatcttcca
     2761 gttcagactc tagtgcagag tcagacgacc aatgcttgat gtcatcgagc accccggagt
     2821 gtgctgcggg tttcttaaag acagtaggcc tttttgcagg aagaggtcgt ccaggcccag
     2881 ggctgtcatc acagactgca ggtgctgctg gatggaggcg ttctggctca aatggtggtg
     2941 gacaggctcc tggtgcttct cccagtgtgt ctctccctgc tagtttagga agaggatggg
     3001 gtagagaaga gaaccttttt tcttggaagg gagctaaggg acggggcatg cgggggagag
     3061 gtcgaggacg agggcatcct gtttcctgtg ttgtaaatag aagcactgac aaccagaggc
     3121 aacagcaatt aaatgacgtg gtaaaaaatt catctactat tatccagaat ccagtagaga
     3181 cacccaagaa ggactatagt ctgttaccac tgttagcagc tgcccctcaa gttggagaaa
     3241 agattgcatt taagcttttg gagctaacat ccagttactc tcctgatgtc tctgactacg
     3301 aggaaggaag aatattaagc cacaatccag agacccagca agtagatata gaaattcttt
     3361 catccttacc tgccttgaga gaacctggga aatttgattt agtttatcac aatgaaaatg
     3421 gagctgaggt agtggagtac gctgtgacac aggagagcaa gatcactgta ttttggaaag
     3481 agttgattga cccaagactg attattgaat ctccaagtaa cacatcaagt acagaacctg
     3541 cctaagagag ctaagcggcc gcgactctag atcataatca gccataccac atttgtagag
     3601 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     3661 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3721 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3781 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     3841 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     3901 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     3961 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     4021 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     4081 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     4141 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     4201 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     4261 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4321 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4381 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     4441 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4501 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4561 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4621 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4681 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4741 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4801 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4861 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4921 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     4981 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5041 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5101 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5161 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5221 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5281 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     5341 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5401 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5461 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5521 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5581 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     5641 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5701 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     5761 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     5821 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     5881 ggctggcact ctgtcgatac cccaccgaga ccccattggg gtcaatacgc ccgcgtttct
     5941 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     6001 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     6061 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6121 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6181 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6241 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6301 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6361 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6421 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6481 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6541 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6601 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6661 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6721 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6781 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6841 cctgcgttat cc
//
