LOCUS       sequence-475413-        6672 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing fluorescent Grx1-roGFP2 fusion protein in mammalian cells
            with localization in paraspeckles..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6672)
  TITLE     Rivera-Fuentes plasmids March 2025
REFERENCE   2  (bases 1 to 6672)
  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-PSPC1
FEATURES             Location/Qualifiers
     source          1..6672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(831..1541,1542..1544))
                     /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(873..894)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1222..1241
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1481..1502
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1928..1937
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2020..2223)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2049..2069)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2224..2527
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2702..3290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2782..2801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3619..3666
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3691..3710
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3898..4692)
                     /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             complement(3898..4692)
                     /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(4009..4028)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4619..4638
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4727..5084)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4741..4876
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4893..5084
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5041..5061
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5086..5190)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5217..5672
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5304..5323)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5514..5535
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5679..5800
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5690..5709)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5744..5763
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6140..6151)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
ORIGIN
        1 gatgcatact caaattcaaa tgtcccaggt tgagcaaaac gtggtggttg ttctctttcc
       61 ttatgatatt gttgagtttt ctgcatcagc ttctctggca agccatcttc atcatcaaac
      121 tgctccatgg gttccacaat gactggacga ggggtcgttg ttagcaagaa tgccccatca
      181 ccacatcttt ccagagcctt tcgtgcagga ggttttgctg caaactctac aaaacctttt
      241 cctgtagctc taccgcgatc atccacaacc acaacagctt tctctactgg accaaactga
      301 gaaaatgctt gctctagcag ctcattggaa acaactggag aaaggttctt gacagtcaag
      361 gctgctccat gtgtagcgaa gcgaatccgt agaggtctgc tcttgagaat ggtgccgtcc
      421 agctctgctt ttgcaatttc agccagggtt ctggattcca agcggatgaa gccgaagcca
      481 cggtcccggt tgatgaagac ttcgctgggc tcgccatagc gttcgaagag cctcttgaag
      541 tcctcctccg tgatgtcggt gggcagattt cccacgaaga ggcggcagcg ctgcgtgtac
      601 gtcttctcgc ccggcttgag gaaactcttg atgtcgatag tgaaccccat ctcctcgtcc
      661 gggtggtcct ctggaggcgc gggcgcgggc ggtgccggct ccccggcaag agcgagcgcc
      721 atggctgccg cggccgccgg ctcgctctcg cccaccgcgg actccagggc gcgaaggcgg
      781 gccgggtttt tctcaatgcg cacttgcttc aggtttcctc ttaacatcat cttgtacagc
      841 tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg
      901 cgcttctcgt tggggtcttt gctcagggcg gagcaggtgc tcaggtagtg gttgtcgggc
      961 agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg
     1021 ctgccgtcct cgatgttgtg gcggatcttg aagttcacct tgatgccgtt cttctgcttg
     1081 tcggccatga tatagacgtt gtggcagttg tagttgtact ccagcttgtg ccccaggatg
     1141 ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc
     1201 tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc
     1261 tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg
     1321 gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc
     1381 acgggcagct tgccggtggt ggagatgaac ttcagggtca gcttgccgta ggtggcatcg
     1441 ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc
     1501 aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcacgaattc ctcgggcagc
     1561 ttcctggcct tcttctccag ctccttgatc ttctcgccgg cctcctcctc agtctgcaga
     1621 gctccaatct gctttagccg cgtcagcagt tccccactct gttgcaaaga gactagatca
     1681 ctgcatccgc ctatacaatc tttaccaata aagactcgag gcaccgttct tgctcccgtg
     1741 agctgttgca aataatcttg aatctcgtta gtgtggttgg tggctgtgat atcgacaaat
     1801 tccagaagcc cttgtttgat gggcaattga ctgaggatct cttgggccct cctgcagtac
     1861 gggcaggtgg gcttgatgaa cacaaccacc ttcccaggct ggattttgca gttcacaaac
     1921 tcttgagcca tggtggcggt accgtcgact gcagaattcg aagcttgagc tcgagatctg
     1981 agtccggtag cgctagcgga tctgacggtt cactaaacca gctctgctta tatagacctc
     2041 ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg
     2101 aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact
     2161 tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat
     2221 caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa
     2281 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg
     2341 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
     2401 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
     2461 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta
     2521 tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg attactatta
     2581 ataactaatg catggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2641 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2701 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2761 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2821 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2881 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2941 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3001 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3061 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3121 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3181 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3241 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3301 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3361 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3421 aatcaatcta aagtatatat gagtaacctg aggctatggc agggcctgcc gccccgacgt
     3481 tggctgcgag ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa
     3541 cgcgggcgta ttgaccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg
     3601 accgaacccc gcgtttatga acaaacgacc caacaccgtg cgttttattc tgtcttttta
     3661 ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc
     3721 ccctagggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg
     3781 cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat
     3841 ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca
     3901 gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc
     3961 gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt
     4021 agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc
     4081 agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac
     4141 gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag
     4201 cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg
     4261 tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt
     4321 atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga
     4381 tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt
     4441 gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc
     4501 tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg
     4561 gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc
     4621 ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc
     4681 ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcgatctttg caaaagccta
     4741 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     4801 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     4861 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
     4921 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
     4981 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     5041 accctaactg acacacattc cacagctggt tctttccgcc tcaggactct tcctttttca
     5101 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5161 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc
     5221 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5281 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5341 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5401 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     5461 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     5521 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     5581 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5641 gaattttaac aaaatattaa cgcttacaat ttacgcctta agatacattg atgagtttgg
     5701 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     5761 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     5821 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     5881 caaatgtggt atggctgatt atgatctaga gtcgcggccg cttagctctc ttaatatcta
     5941 cggcgcttat tagggccttc aaagttgccc ccttgacttc ctctaccaaa gccaccagga
     6001 ccaccactca caggacctac accactcatt ggtgcttgag gggtttcaga acctgttcta
     6061 ctccccatag gtgaacccat ctgagatggt ggtccttgag gaaatctgtc attgtgcact
     6121 gctccattat ctggcatcat tggagttccc atatttgcgg ctccttctgg tcccatggca
     6181 ggaccaggac ccattggtgg gccaggtata gttgctctgt tgttcatatt catacccatc
     6241 attggaggag gaccttggtt accagcaggg gctgggctaa acgcatctcc catgtttatt
     6301 gctccacggg gacccatatc acccattctc atttcctgtt ctctattttc catgtagttt
     6361 ggcttaaagc cctcttgctg tcgcctcagt tcctcctgtt ctctgtgtcg gatcatttct
     6421 tcctcacgcc gccgatgctc ctcttcatgt cttagttgta tttgcttccg tttttgcaac
     6481 tcttggtttc tgagttcttc caagcgtctg agttcttctt gacgcctcat tagatcttgc
     6541 ctcattagca ttaattggtg ttcatgccta gctgcttcca tttctgcctc cagtttctct
     6601 ttggcttctc tgatgtttct atcaacctgc tcacgctgct gcttttccat ttcatcaaga
     6661 gccttccatc ga
//
