LOCUS       sequence-444231-        5182 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mitochondrion-localized BmTyr.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5182)
  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 5182)
  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: 2COX8-BmTyr
FEATURES             Location/Qualifiers
     source          1..5182
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     224..243
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(332..689)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      346..481
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        498..689
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     646..666
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(691..795)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      822..1277
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(909..928)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1119..1140
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1284..1405
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1295..1314)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1349..1368
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1530..1553)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(2493..2567)
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             complement(2607..2681)
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     misc_feature    2704..2784
                     /label=MCS
                     /note="multiple cloning site"
     promoter        complement(2807..3010)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2836..2856)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3011..3314
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3489..4077)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3569..3588)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4406..4453
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4478..4497
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(join(4685..5182,1..297))
                     /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             complement(join(4685..5182,1..297))
                     /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(4796..4815)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
       61 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag
      121 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
      181 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
      241 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg
      301 aaacgatcct catcctgtct cttgatcgat ctttgcaaaa gcctaggcct ccaaaaaagc
      361 ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa
      421 aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga
      481 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac
      541 ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca
      601 tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca
      661 cattccacag ctggttcttt ccgcctcagg actcttcctt tttcaatatt attgaagcat
      721 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
      781 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc
      841 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      901 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      961 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     1021 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     1081 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     1141 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     1201 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     1261 attaacgctt acaatttacg ccttaagata cattgatgag tttggacaaa ccacaactag
     1321 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1381 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1441 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     1501 tgattatgat ctagagtcgc ggccgctcac ttatcgtcgt cgtccttata gtcagatgac
     1561 ctcttggatt tgcggagttc gatatcgtag acgtatccca gtttccgatg gttcatcacg
     1621 tcctccggtg tagtattcca cgggtacatt gggtcacgaa agttctgacc gaatgggcca
     1681 ttcttcatgg gctggtaatt ctggttgcga tgtatgatct gccaaacggc ccagattcgg
     1741 tccacattgg catgatggag gaagaacacg ggatcatttg gtgcagtggg gactacgccc
     1801 atctgtccgc caacccatct gtggactctg ttgtgcagct gaggcccgtt aatgaacccc
     1861 tcaagctgat tcctgaagga gttctgggaa gtcatatccc agggtggtgt gtcatactgg
     1921 gtaatcttca gtgcgttgag tacatcatcc cgagtcggca gtgtgggtgc ttctttggtg
     1981 gctccgaaat tccttttcag accgcctgaa gggttgcctt gctcatcgat ggtggtccat
     2041 ctaccagcag caaaaggtcc ggtatcaacg atgaagtcct tgatagggtt cccattgcct
     2101 cccataaagt cggcagacca aatctggctt tggcttggat cctgcatttg cgcatctgtc
     2161 tcccactccc aatagggcaa cgtcacttcg gggtttatgc tttgcaagtc gcgttcaaac
     2221 cggagcagat attcccggtg ccaaggaaga aaggcagaac tcatgtgagc ggcattgcga
     2281 tcgctccctg gaggagtgtg gaatttccct gccgcaccgt gccaagcgat gtacctgtca
     2341 taaatgcctt tctccttcag aatcagcacc gttctcacaa agtctcgttt ctctgtatcg
     2401 gtcagatgca ggacattctt tctcacacgg tacttgttac tgatctgatg ggtggagccg
     2461 cccaacttgc cctccggtgg caacgaatgg atcttggcgc gcggcactgg gagccgccgg
     2521 gccgagcctg tcaagccccg cagcagcagc ggcgtcagga cggacatgga gccgcccaac
     2581 ttgccctccg gtggcaacga atggatcttg gcgcgcggca ctgggagccg ccgggccgag
     2641 cctgtcaagc cccgcagcag cagcggcgtc aggacggaca tggtggctag cggatcggtg
     2701 gcgaccggtg gatcccgggc ccgcggtacc gtcgactgca gaattcgaag cttgagctcg
     2761 agatctgagt ccggtagcgc tagcggatct gacggttcac taaaccagct ctgcttatat
     2821 agacctccca ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac
     2881 attttggaaa gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt
     2941 ggagacttgg aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa
     3001 accgcatcac catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt
     3061 cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa
     3121 tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta
     3181 aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg
     3241 tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg
     3301 taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt
     3361 actattaata actaatgcat ggcggtaata cggttatcca cagaatcagg ggataacgca
     3421 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3481 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3541 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3601 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3661 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3721 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3781 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3841 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3901 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3961 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4021 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4081 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4141 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4201 agttttaaat caatctaaag tatatatgag taacctgagg ctatggcagg gcctgccgcc
     4261 ccgacgttgg ctgcgagccc tgggccttca cccgaacttg gggggtgggg tggggaaaag
     4321 gaagaaacgc gggcgtattg gccccaatgg ggtctcggtg gggtatcgac agagtgccag
     4381 ccctgggacc gaaccccgcg tttatgaaca aacgacccaa caccgtgcgt tttattctgt
     4441 ctttttattg ccgtcatagc gcgggttcct tccggtattg tctccttccg tgtttcagtt
     4501 agcctccccc tagggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc
     4561 cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa
     4621 tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc
     4681 ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg
     4741 cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat
     4801 cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga
     4861 tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg
     4921 tcacgacgag atcctcgccg tcgggcatgc tcgccttgag cctggcgaac agttcggctg
     4981 gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc
     5041 gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat
     5101 caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa
     5161 ggtgagatga caggagatcc tg
//
