LOCUS       sequence-444229-        6056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  H2B-BmTyr fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6056)
  AUTHORS   Zhu H, Oh JH, Matsuda Y, Mino T, Ishikawa M, Nakamura H, Tsujikawa 
            M, Nonaka H, Hamachi I
  TITLE     Tyrosinase-Based Proximity Labeling in Living Cells and In Vivo.
  JOURNAL   J Am Chem Soc. 2024 Mar 20;146(11):7515-7523. doi: 
            10.1021/jacs.3c13183. Epub 2024 Mar 6.
  PUBMED    38445591
REFERENCE   2  (bases 1 to 6056)
  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/jacs.3c13183"; journalName: "J Am Chem Soc"; date: 
            "2024-03-20- 20"; volume: "146"; issue: "11"; pages: "7515-7523"
COMMENT     Sequence Label: mCherry-BmTyr-H2B
FEATURES             Location/Qualifiers
     source          1..6056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    548..669
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(585..604)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     639..658
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(676..1131)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(813..834)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1025..1044
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1158..1262
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1264..1621
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1264..1455
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1287..1307)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1472..1607
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1656..2450
                     /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             1656..2450
                     /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(1710..1729)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2320..2339
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2638..2657)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2682..2729
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3058..3646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3547..3566
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3821..4124
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4125..4328
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4279..4299
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4367..4376
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4373..5080
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4518..4536)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4786..4805
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             join(6053..6056,1..380)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKDGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSAKV"
ORIGIN
        1 cagagccagc gaagtctgct cccgccccga aaaagggctc caagaaggcg gtgactaagg
       61 cgcagaagaa agacggcaag aagcgcaagc gcagccgcaa ggagagctat tccatctatg
      121 tgtacaaggt tctgaagcag gtccaccctg acaccggcat ttcgtccaag gccatgggca
      181 tcatgaattc gtttgtgaac gacattttcg agcgcatcgc aggtgaggct tcccgcctgg
      241 cgcattacaa caagcgctcg accatcacct ccagggagat ccagacggcc gtgcgcctgc
      301 tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg tactaaggcc gtcaccaagt
      361 acaccagcgc taaggtttaa aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg
      421 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact
      481 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
      541 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
      601 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
      661 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
      721 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
      781 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
      841 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
      901 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
      961 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     1021 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     1081 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     1141 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     1201 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     1261 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     1321 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     1381 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     1441 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     1501 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     1561 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     1621 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     1681 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     1741 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     1801 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     1861 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     1921 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     1981 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     2041 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     2101 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     2161 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     2221 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     2281 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     2341 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     2401 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     2461 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     2521 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     2581 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     2641 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     2701 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     2761 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     2821 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     2881 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     2941 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3001 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3061 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3121 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3181 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3241 gaactctgta tcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3301 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3361 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3421 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3481 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3541 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3601 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3661 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3721 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa
     3781 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     3841 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     3901 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     3961 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     4021 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     4081 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     4141 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     4201 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     4261 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     4321 gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggtcgcca ccatggtaag
     4381 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
     4441 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     4501 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
     4561 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
     4621 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     4681 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     4741 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     4801 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     4861 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
     4921 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     4981 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     5041 gggccgccac tccaccggcg gcatggacga gctgtacaag agtgctggtg gtatgagtaa
     5101 caagtaccgt gtgagaaaga atgtcctgca tctgaccgat acagagaaac gagactttgt
     5161 gagaacggtg ctgattctga aggagaaagg catttatgac aggtacatcg cttggcacgg
     5221 tgcggcaggg aaattccaca ctcctccagg gagcgatcgc aatgccgctc acatgagttc
     5281 tgcctttctt ccttggcacc gggaatatct gctccggttt gaacgcgact tgcaaagcat
     5341 aaaccccgaa gtgacgttgc cctattggga gtgggagaca gatgcgcaaa tgcaggaccc
     5401 aagccaaagc cagatttggt ctgccgactt tatgggaggc aatgggaacc ctatcaagga
     5461 cttcatcgtt gataccggac cttttgctgc tggtagatgg accaccatcg atgagcaagg
     5521 caacccttca ggcggtctga aaaggaattt cggagccacc aaagaagcac ccacactgcc
     5581 gactcgggat gatgtactca acgcactgaa gattacccag tatgacacac caccctggga
     5641 tatgacttcc cagaactcct tcaggaatca gcttgagggg ttcattaacg ggcctcagct
     5701 gcacaacaga gtccacagat gggttggcgg acagatgggc gtagtcccca ctgcaccaaa
     5761 tgatcccgtg ttcttcctcc atcatgccaa tgtggaccga atctgggccg tttggcagat
     5821 catacatcgc aaccagaatt accagcccat gaagaatggc ccattcggtc agaactttcg
     5881 tgacccaatg tacccgtgga atactacacc ggaggacgtg atgaaccatc ggaaactggg
     5941 atacgtctac gatatcgaac tccgcaaatc caagaggtca tctagtgctg gtggtagtgc
     6001 tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggttctcgag ctatgc
//
