LOCUS       sequence-254783-        5115 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Outer Mitochondrial Membrane, Excitation: 399, 
            Emission: 456.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5115)
  AUTHORS   Subach OM, Cranfill PJ, Davidson MW, Verkhusha VV
  TITLE     An enhanced monomeric blue fluorescent protein with the high 
            chemical stability of the chromophore.
  JOURNAL   PLoS One. 2011;6(12):e28674. Epub 2011 Dec 8.
  PUBMED    22174863
REFERENCE   2  (bases 1 to 5115)
  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"; journalName: "PLoS One.";
            date: "2011"; volume: "6(12)"; pages: "e28674. Epub 2011 Dec 8"
COMMENT     Sequence Label: mTagBFP2-TOMM20-N-10
FEATURES             Location/Qualifiers
     source          1..5115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             73..867
                     /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             73..867
                     /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(127..146)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     737..756
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1055..1074)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1099..1146
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1475..2063
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1964..1983
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2238..2541
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2542..2745
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2696..2716
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2774..2783
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3239..3248
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3245..3955
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     polyA_signal    4080..4201
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4117..4136)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4171..4190
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4208..4663)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4345..4366)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4557..4576
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4690..4794
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(4796..5115,1..38)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4796..4987
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4819..4839)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(5004..5115,1..24)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
       61 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
      121 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
      181 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
      241 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
      301 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
      361 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
      421 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
      481 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
      541 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
      601 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
      661 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
      721 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
      781 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
      841 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
      901 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
      961 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     1021 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     1081 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     1141 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     1201 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     1261 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     1321 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     1381 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     1441 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     1501 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     1561 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     1621 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     1681 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     1741 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     1801 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     1861 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     1921 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     1981 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     2041 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     2101 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     2161 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
     2221 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     2281 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     2341 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     2401 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     2461 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     2521 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     2581 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     2641 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     2701 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
     2761 cagatccgct agcgccacca tggtgggtcg gaacagcgcc atcgccgccg gtgtatgcgg
     2821 ggcccttttc attgggtact gcatctactt cgaccgcaaa agacgaagtg accccaactt
     2881 caagaacagg cttcgagaac gaagaaagaa acagaagctt gccaaggaga gagctgggct
     2941 ttccaagtta cctgacctta aagatgctga agctgttcag aagttcttcc ttgaagaaat
     3001 acagcttggt gaagagttac tagctcaagg tgaatatgag aagggcgtag accatctgac
     3061 aaatgcaatt gctgtgtgtg gacagccaca gcagttactg caggtcttac agcaaactct
     3121 tccaccacca gtgttccaga tgcttctgac taagctccca acaattagtc agagaattgt
     3181 aagtgctcag agcttggctg aagatgatgt ggaaggcggt agcggggatc caccggtcgc
     3241 caccatggtg tctaagggcg aagagctgat taaggagaac atgcacatga agctgtacat
     3301 ggagggcacc gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta
     3361 cgagggcacc cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt
     3421 cgacatcctg gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg
     3481 catccccgac ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac
     3541 atacgaagac gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct
     3601 catctacaac gtcaagatca gaggggtgaa cttcacatcc aacggccctg tgatgcagaa
     3661 gaaaacactc ggctgggagg ccttcaccga gacgctgtac cccgctgacg gcggcctgga
     3721 aggcagaaac gacatggccc tgaagctcgt gggcgggagc catctgatcg caaacgccaa
     3781 gaccacatat agatccaaga aacccgctaa gaacctcaag atgcctggcg tctactatgt
     3841 ggactacaga ctggaaagaa tcaaggaggc caacaacgag acctacgtcg agcagcacga
     3901 ggtggcagtg gccagatact gcgacctccc tagcaaactg gggcacaagc ttaattaagc
     3961 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     4021 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     4081 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4141 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4201 aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     4261 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     4321 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     4381 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     4441 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     4501 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     4561 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     4621 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     4681 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4741 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     4801 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4861 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4921 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4981 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     5041 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     5101 agctattcca gaagt
//
