LOCUS       sequence-362760-        6219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses fluorescent HaloTag7 fusion protein in mammalian cells 
            with nuclear localization..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6219)
  AUTHORS   Emmert S, Quargnali G, Thallmair S, Rivera-Fuentes P
  TITLE     A locally activatable sensor for robust quantification of organellar
            glutathione.
  JOURNAL   Nat Chem. 2023 Oct;15(10):1415-1421. doi: 
            10.1038/s41557-023-01249-3. Epub 2023 Jun 15.
  PUBMED    37322101
REFERENCE   2  (bases 1 to 6219)
  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"; doi: 
            "10.1038/s41557-023-01249-3"; journalName: "Nat Chem"; date: 
            "2023-10"; volume: "15"; issue: "10"; pages: "1415-1421"
COMMENT     Sequence Label: H2B-HT-emiRFP703
FEATURES             Location/Qualifiers
     source          1..6219
                     /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
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             1086..1970
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     regulatory      1983..1992
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3043..3164
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3080..3099)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3134..3153
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3171..3626)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3308..3329)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3520..3539
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3653..3757
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3759..4116
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3759..3950
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3782..3802)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3967..4102
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4151..4945
                     /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             4151..4945
                     /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(4205..4224)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4815..4834
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5133..5152)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5177..5224
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5553..6141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6042..6061
                     /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 tccaaggcca 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 gcagcgaaat cggtactggc tttccattcg acccccatta tgtggaagtc ctgggcgagc
     1141 gcatgcacta cgtcgatgtt ggtccgcgcg atggcacccc tgtgctgttc ctgcacggta
     1201 acccgacctc ctcctacgtg tggcgcaaca tcatcccgca tgttgcaccg acccatcgct
     1261 gcattgctcc agacctgatc ggtatgggca aatccgacaa accagacctg ggttatttct
     1321 tcgacgacca cgtccgcttc atggatgcct tcatcgaagc cctgggtctg gaagaggtcg
     1381 tcctggtcat tcacgactgg ggctccgctc tgggtttcca ctgggccaag cgcaatccag
     1441 agcgcgtcaa aggtattgca tttatggagt tcatccgccc tatcccgacc tgggacgaat
     1501 ggccagaatt tgcccgcgag accttccagg ccttccgcac caccgacgtc ggccgcaagc
     1561 tgatcatcga tcagaacgtt tttatcgagg gtacgctgcc gatgggtgtc gtccgcccgc
     1621 tgactgaagt cgagatggac cattaccgcg agccgttcct gaatcctgtt gaccgcgagc
     1681 cactgtggcg cttcccaaac gagctgccaa tcgccggtga gccagcgaac atcgtcgcgc
     1741 tggtcgaaga atacatggac tggctgcacc agtcccctgt cccgaagctg ctgttctggg
     1801 gcaccccagg cgttctgatc ccaccggccg aagccgctcg cctggccaaa agcctgccta
     1861 actgcaaggc tgtggacatc ggcccgggtc tgaatctgct gcaagaagac aacccggacc
     1921 tgatcggcag cgagatcgcg cgctggctgt ccacgctcga gatttccggc gatccaccgg
     1981 tcgccaccat ggcggaaggc tccgtcgcca ggcagcctga cctcttgacc tgcgaacatg
     2041 aagagatcca cctcgccggc tcgatccagc cgcatggcgc gcttctggtc gtcagcgaac
     2101 atgatcatcg cgtcatccag gccagcgcca acgccgcgga atttctgaat ctcggaagcg
     2161 tactcggcgt tccgctcgcc gagatcgacg gcgatctgtt gatcaagatc ctgccgcatc
     2221 tcgatcccac cgccgaaggc atgccggtcg cggtgcgctg ccggatcggc aatccctcta
     2281 cggagtactg cggtctgatg catcggcctc cggaaggcgg gctgatcatc gaactcgaac
     2341 gtgccggccc gtcgatcgat ctgtcaggca cgctggcgcc ggcgctggag cggatccgca
     2401 cggcgggttc actgcgcgcg ctgtgcgatg acaccgtgct gctgtttcag cagtgcaccg
     2461 gctacgaccg ggtgatggtg tatcgtttcg atgagcaagg ccacggcctg gtattctccg
     2521 agtgccatgt gcctgggctc gaatcctatt tcggcaaccg ctatccgtcg tcgctggtcc
     2581 cgcagatggc gcggcagctg tacgtgcggc agcgcgtccg cgtgctggtc gacgtcacct
     2641 atcagccggt gccgctggag ccgcggctgt cgccgctgac cgggcgcgat ctcgacatgt
     2701 cgggctgctt cctgcgctcg atgtcgccga tccatctgca gttcctgaag gacatgggcg
     2761 tgcgcgccac cctggcggtg tcgctggtgg tcggcggcaa gctgtggggc ctggttgtct
     2821 gtcaccatta tctgccgcgc ttcatccgtt tcgagctgcg ggcgatctgc aaacggctcg
     2881 ccgaaaggat cgcgacgcgg atcaccgcgc ttgagagcta agcggccgcg actctagatc
     2941 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     3001 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     3061 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3121 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     3181 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     3241 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     3301 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     3361 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     3421 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     3481 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     3541 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     3601 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     3661 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3721 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     3781 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3841 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3901 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3961 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4021 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     4081 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     4141 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4201 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4261 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4321 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4381 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4441 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4501 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4561 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4621 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     4681 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4741 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4801 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4861 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4921 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4981 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5041 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5101 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     5161 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     5221 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     5281 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     5341 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     5401 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5461 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5521 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5581 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5641 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5701 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5761 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5821 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5881 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5941 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6001 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6061 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6121 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6181 tgctggcctt ttgctcacat gttctttcct gcgttatcc
//
