LOCUS       sequence-256754-        4774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Plasma Membrane, Excitation: 399, Emission: 456.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4774)
  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 4774)
  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-Farnesyl-5
FEATURES             Location/Qualifiers
     source          1..4774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        93..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        397..600
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     551..571
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      639..648
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             645..1355
                     /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    1594..1715
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1631..1650)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1685..1704
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1722..2177)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1859..1880)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2071..2090
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2204..2308
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2310..2667
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2310..2501
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2333..2353)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2518..2653
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2702..3496
                     /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             2702..3496
                     /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(2756..2775)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3366..3385
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3684..3703)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3728..3775
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4104..4692
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4593..4612
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg
       61 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      121 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      181 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      241 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      301 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      361 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      421 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      481 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      541 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      601 ggtttagtga accgtcagat ccgctagcgc taccggtcgc caccatggtg tctaagggcg
      661 aagagctgat taaggagaac atgcacatga agctgtacat ggagggcacc gtggacaacc
      721 atcacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga
      781 gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctactagct
      841 tcctctacgg cagcaagacc ttcatcaacc acacccaggg catccccgac ttcttcaagc
      901 agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc
      961 tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac gtcaagatca
     1021 gaggggtgaa cttcacatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg
     1081 ccttcaccga gacgctgtac cccgctgacg gcggcctgga aggcagaaac gacatggccc
     1141 tgaagctcgt gggcgggagc catctgatcg caaacgccaa gaccacatat agatccaaga
     1201 aacccgctaa gaacctcaag atgcctggcg tctactatgt ggactacaga ctggaaagaa
     1261 tcaaggaggc caacaacgag acctacgtcg agcagcacga ggtggcagtg gccagatact
     1321 gcgacctccc tagcaaactg gggcacaagc ttaattccgg actcagatct aagctgaacc
     1381 ctcctgatga gagtggcccc ggctgcatga gctgcaagtg tgtgctctcc tgagaattct
     1441 gcagtcgacg gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc
     1501 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     1561 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     1621 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1681 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     1741 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     1801 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     1861 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     1921 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     1981 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2041 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2101 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2161 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2221 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2281 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     2341 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2401 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     2461 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     2521 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     2581 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     2641 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     2701 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     2761 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     2821 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     2881 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     2941 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3001 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3061 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3121 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3181 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     3241 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3301 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3361 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3421 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3481 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     3541 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3601 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3661 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     3721 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     3781 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     3841 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     3901 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     3961 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4021 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4081 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4141 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4201 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4261 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4321 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4381 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4441 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4501 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4561 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4621 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4681 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4741 tttgctcaca tgttctttcc tgcgttatcc cctg
//
