LOCUS       sequence-426765-        4500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MiniR genetically encoded tag introduced after H2B; to be used with 
            a MiniE probe peptide..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4500)
  AUTHORS   Suyama A, Devlin KL, Macias-Contreras M, Doh JK, Shinde U, Beatty KE
  TITLE     Orthogonal Versatile Interacting Peptide Tags for Imaging Cellular 
            Proteins.
  JOURNAL   Biochemistry. 2023 Jun 6;62(11):1735-1743. doi: 
            10.1021/acs.biochem.2c00712. Epub 2023 May 11.
  PUBMED    37167569
REFERENCE   2  (bases 1 to 4500)
  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.2c00712"; journalName: "Biochemistry"; date: 
            "2023-06-6- 6"; volume: "62"; issue: "11"; pages: "1735-1743"
COMMENT     Sequence Label: H2B-MiniR
FEATURES             Location/Qualifiers
     source          1..4500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     73..92
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        347..650
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        651..854
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     805..825
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             943..1320
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     polyA_signal    1574..1695
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1611..1630)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1665..1684
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1702..2157)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1839..1860)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2051..2070
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2184..2288
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2290..2647
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2290..2481
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2313..2333)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2498..2633
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2682..3476
                     /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             2682..3476
                     /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(2736..2755)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3346..3365
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3664..3683)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3708..3755
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(4084..4500,1..172)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
       61 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      121 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      181 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      241 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
      301 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      361 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      421 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      481 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      541 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      601 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      661 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      721 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      781 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      841 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct
      901 cgagctcaag cttcgaattc tgcagtcgac ggtaccgcca ccatgccaga gccagcgaag
      961 tctgctcccg ccccgaaaaa gggctccaag aaggcggtga ctaaggcgca gaagaaaggc
     1021 ggcaagaagc gcaagcgcag ccgcaaggag agctattcca tctatgtgta caaggttctg
     1081 aagcaggtcc accctgacac cggcatttcg tccaaggcca tgggcatcat gaattcgttt
     1141 gtgaacgaca ttttcgagcg catcgcaggt gaggcttccc gcctggcgca ttacaacaag
     1201 cgctcgacca tcacctccag ggagatccag acggccgtgc gcctgctgct gcctggggag
     1261 ttggccaagc acgccgtgtc cgagggtact aaggccatca ccaagtacac cagcgctaag
     1321 gatccaatgc tcgaaatccg tgttgcgttc ctgcgtcagc gtaacaccgc gctgcgtacc
     1381 gaagttgcgg aactggaaca ggaagttcag cgtctggaaa accgttacgg tccgctgccg
     1441 gtcgccacct aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     1501 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     1561 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     1621 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     1681 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     1741 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     1801 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     1861 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1921 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     1981 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     2041 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     2101 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     2161 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2221 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2281 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     2341 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2401 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2461 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     2521 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     2581 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     2641 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     2701 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     2761 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     2821 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     2881 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     2941 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3001 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3061 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3121 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3181 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     3241 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3301 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3361 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3421 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     3481 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     3541 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     3601 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     3661 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     3721 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     3781 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     3841 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     3901 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     3961 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4021 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4081 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4141 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4201 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4261 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4321 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4381 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4441 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
//
