LOCUS       sequence-366155-        5217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MiniE genetically encoded tag on H2B-mEmerald to be used with MiniR 
            labeled with a reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5217)
  AUTHORS   Doh JK, Tobin SJ, Beatty KE
  TITLE     MiniVIPER Is a Peptide Tag for Imaging and Translocating Proteins in
            Cells.
  JOURNAL   Biochemistry. 2020 Aug 25;59(33):3051-3059. doi: 
            10.1021/acs.biochem.0c00526. Epub 2020 Aug 17.
  PUBMED    32786411
REFERENCE   2  (bases 1 to 5217)
  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: 
            "10.1021/acs.biochem.0c00526"; journalName: "Biochemistry"; date: 
            "2020-08-25- 25"; volume: "59"; issue: "33"; pages: "3051-3059"
COMMENT     Sequence Label: H2B-MiniE-mEm
FEATURES             Location/Qualifiers
     source          1..5217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        154..457
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        458..661
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     612..632
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             750..1127
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1251..1260
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1257..1259,1260..1262,1263..1976)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1302..1323)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1563..1582)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1910..1931
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2098..2219
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2135..2154)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2189..2208
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2226..2681)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2363..2384)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2575..2594
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2708..2812
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2814..3171
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2814..3005
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2837..2857)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3022..3157
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3206..4000
                     /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             3206..4000
                     /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(3260..3279)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3870..3889
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4188..4207)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4232..4279
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4608..5196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5097..5116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
       61 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg
      121 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      181 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      241 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      301 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      361 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      421 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      481 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      541 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      601 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      661 tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatctcga gctcaagctt
      721 cgaattctgc agtcgacggt accgccacca tgccagagcc agcgaagtct gctcccgccc
      781 cgaaaaaggg ctccaagaag gcggtgacta aggcgcagaa gaaaggcggc aagaagcgca
      841 agcgcagccg caaggagagc tattccatct atgtgtacaa ggttctgaag caggtccacc
      901 ctgacaccgg catttcgtcc aaggccatgg gcatcatgaa ttcgtttgtg aacgacattt
      961 tcgagcgcat cgcaggtgag gcttcccgcc tggcgcatta caacaagcgc tcgaccatca
     1021 cctccaggga gatccagacg gccgtgcgcc tgctgctgcc tggggagttg gccaagcacg
     1081 ccgtgtccga gggtactaag gccatcacca agtacaccag cgctaaggat ccaatgctgg
     1141 agatcgaggc cgccttcctc gagagggaga acaccgccct ggagaccagg gtggccgagc
     1201 tgaggcagag ggtgcagagg ctgaggaacg agtacggccc tctgccggtc gccaccatgg
     1261 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1321 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1381 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1441 tgaccacctt gacctacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc
     1501 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1561 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1621 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1681 tggagtacaa ctacaacagc cacaaggtct atatcaccgc cgacaagcag aagaacggca
     1741 tcaaggtgaa cttcaagacc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     1801 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1861 tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1921 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagcgg
     1981 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2041 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2101 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     2161 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     2221 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2281 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     2341 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     2401 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     2461 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     2521 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     2581 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     2641 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     2701 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2761 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     2821 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     2881 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     2941 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3001 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3061 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3121 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     3181 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3241 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3301 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3361 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     3421 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3481 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     3541 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     3601 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     3661 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     3721 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     3781 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     3841 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     3901 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     3961 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4021 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4081 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4141 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     4201 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     4261 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     4321 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     4381 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     4441 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4501 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4561 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4621 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4681 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4741 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4801 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4861 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4921 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4981 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5041 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5101 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5161 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttt
//
