LOCUS       sequence-444228-        4892 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  non-targeted BmTyr throughout the cytoplasm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4892)
  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 4892)
  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: BmTyr-Flag
FEATURES             Location/Qualifiers
     source          1..4892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     432..451
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        706..1009
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1010..1213
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1164..1184
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2177..2200
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2325..2446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2362..2381)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2416..2435
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2453..2908)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2590..2611)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2802..2821
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2935..3039
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3041..3398
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3041..3232
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3064..3084)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3249..3384
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3433..4227
                     /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             3433..4227
                     /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(3487..3506)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4097..4116
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4415..4434)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4459..4506
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(4835..4892,1..531)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa tgtaactggc
       61 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
      121 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      181 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      241 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      301 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      361 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      421 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      481 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      541 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      601 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
      661 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
      721 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      781 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      841 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      901 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      961 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     1021 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     1081 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     1141 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     1201 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga ctcagatctc
     1261 gagctcaagc ttcgaattcg ccaccatgag taacaagtac cgtgtgagaa agaatgtcct
     1321 gcatctgacc gatacagaga aacgagactt tgtgagaacg gtgctgattc tgaaggagaa
     1381 aggcatttat gacaggtaca tcgcttggca cggtgcggca gggaaattcc acactcctcc
     1441 agggagcgat cgcaatgccg ctcacatgag ttctgccttt cttccttggc accgggaata
     1501 tctgctccgg tttgaacgcg acttgcaaag cataaacccc gaagtgacgt tgccctattg
     1561 ggagtgggag acagatgcgc aaatgcagga tccaagccaa agccagattt ggtctgccga
     1621 ctttatggga ggcaatggga accctatcaa ggacttcatc gttgataccg gaccttttgc
     1681 tgctggtaga tggaccacca tcgatgagca aggcaaccct tcaggcggtc tgaaaaggaa
     1741 tttcggagcc accaaagaag cacccacact gccgactcgg gatgatgtac tcaacgcact
     1801 gaagattacc cagtatgaca caccaccctg ggatatgact tcccagaact ccttcaggaa
     1861 tcagcttgag gggttcatta acgggcctca gctgcacaac agagtccaca gatgggttgg
     1921 cggacagatg ggcgtagtcc ccactgcacc aaatgatccc gtgttcttcc tccatcatgc
     1981 caatgtggac cgaatctggg ccgtttggca gatcatacat cgcaaccaga attaccagcc
     2041 catgaagaat ggcccattcg gtcagaactt tcgtgaccca atgtacccgt ggaatactac
     2101 accggaggac gtgatgaacc atcggaaact gggatacgtc tacgatatcg aactccgcaa
     2161 atccaagagg tcatctgact ataaggacga cgacgataag tgagcggccg cgactctaga
     2221 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     2281 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     2341 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2401 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     2461 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     2521 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     2581 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     2641 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     2701 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     2761 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     2821 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     2881 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     2941 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3001 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     3061 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3121 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3181 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3241 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3301 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3361 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     3421 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3481 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3541 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3601 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3661 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3721 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3781 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3841 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3901 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3961 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4021 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4081 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4141 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4201 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4261 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4321 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4381 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     4441 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     4501 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     4561 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     4621 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     4681 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4741 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4801 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4861 tctgctgctt gcaaacaaaa aaaccaccgc ta
//
