LOCUS       sequence-469939-        5496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing fluorescent Grx1-roGFP2 fusion protein in mammalian cells
            with localization in the nucleus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5496)
  TITLE     Rivera-Fuentes plasmids March 2025
REFERENCE   2  (bases 1 to 5496)
  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: H2B-Grx1-roGFP2
FEATURES             Location/Qualifiers
     source          1..5496
                     /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"
     CDS             693..1070
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSQAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             join(1473..1475,1476..2189)
                     /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(1515..1536)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1776..1795)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2123..2144
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2320..2441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2357..2376)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2411..2430
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2448..2903)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2585..2606)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2797..2816
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2930..3034
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3036..3393
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3036..3227
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3059..3079)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3244..3379
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3428..4222
                     /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             3428..4222
                     /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(3482..3501)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4092..4111
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4410..4429)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4454..4501
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4830..5418
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5319..5338
                     /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 ccatgccaga gccagcgaag tctgctcccg
      721 ccccgaaaaa gggctccaag aaggcggtga ctaaggcgca gaagaaaggc ggcaagaagc
      781 gcaagcgcag ccgcaaggag agctattcca tctatgtgta caaggttctg aagcaggtcc
      841 accctgacac cggcatttcg tcccaggcca tgggcatcat gaattcgttt gtgaacgaca
      901 ttttcgagcg catcgcaggt gaggcttccc gcctggcgca ttacaacaag cgctcgacca
      961 tcacctccag ggagatccag acggccgtgc gcctgctgct gcctggggag ttggccaagc
     1021 acgccgtgtc cgagggtact aaggccatca ccaagtacac tagcgctaag cagtcgaccg
     1081 gcagcatggc tcaagagttt gtgaactgca aaatccagcc tgggaaggtg gttgtgttca
     1141 tcaagcccac ctgcccgtac tgcaggaggg cccaagagat cctcagtcaa ttgcccatca
     1201 aacaagggct tctggaattt gtcgatatca cagccaccaa ccacactaac gagattcaag
     1261 attatttgca acagctcacg ggagcaagaa cggtgcctcg agtctttatt ggtaaagatt
     1321 gtataggcgg atgcagtgat ctagtctctt tgcaacagag tggggaactg ctgacgcggc
     1381 taaagcagat tggagctctg cagactgagg aggaggccgg cgagaagatc aaggagctgg
     1441 agaagaaggc caggaagctg cccgaggaat tcgtgagcaa gggcgaggag ctgttcaccg
     1501 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     1561 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctccacca
     1621 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     1681 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1741 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1801 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1861 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac tgccacaacg
     1921 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     1981 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     2041 acggccccgt gctgctgccc gacaaccact acctgagcac ctgctccgcc ctgagcaaag
     2101 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     2161 ctctcggcat ggacgagctg tacaagtaag agagctaagc ggccgcgact ctagatcata
     2221 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2281 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2341 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2401 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     2461 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     2521 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     2581 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     2641 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     2701 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2761 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     2821 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     2881 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2941 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3001 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     3061 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3121 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3181 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3241 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3301 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3361 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     3421 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3481 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3541 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3601 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3661 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3721 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3781 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3841 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3901 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     3961 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4021 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4081 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4141 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4201 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4261 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4321 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4381 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     4441 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     4501 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     4561 tggggtcaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     4621 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     4681 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4741 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4801 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4861 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4921 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4981 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5041 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5101 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5161 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5221 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5281 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5341 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5401 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5461 tggccttttg ctcacatgtt ctttcctgcg ttatcc
//
