LOCUS       sequence-474325-        5583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Nucleus, Excitation: 399, Emission: 456.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5583)
  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 5583)
  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-PCNA-19-SV40NLS-4
FEATURES             Location/Qualifiers
     source          1..5583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      535..544
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             547..567
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             571..591
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             595..615
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      622..631
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             628..1338
                     /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    2311..2432
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2348..2367)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2402..2421
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2439..2894)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2576..2597)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2788..2807
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2921..3025
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3027..3384
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3027..3218
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3050..3070)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3235..3370
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3419..4213
                     /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             3419..4213
                     /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(3473..3492)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4083..4102
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4401..4420)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4445..4492
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4821..5409
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5310..5329
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagccacc
      541 atggatccaa aaaagaagag aaaggtagat ccaaaaaaga agagaaaggt agatccaaaa
      601 aagaagagaa aggtaccggt cgccaccatg gtgtctaagg gcgaagagct gattaaggag
      661 aacatgcaca tgaagctgta catggagggc accgtggaca accatcactt caagtgcaca
      721 tccgagggcg aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggtggtcgag
      781 ggcggccctc tccccttcgc cttcgacatc ctggctacta gcttcctcta cggcagcaag
      841 accttcatca accacaccca gggcatcccc gacttcttca agcagtcctt ccctgagggc
      901 ttcacatggg agagagtcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac
      961 accagcctcc aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttcaca
     1021 tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg aggccttcac cgagacgctg
     1081 taccccgctg acggcggcct ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg
     1141 agccatctga tcgcaaacgc caagaccaca tatagatcca agaaacccgc taagaacctc
     1201 aagatgcctg gcgtctacta tgtggactac agactggaaa gaatcaagga ggccaacaac
     1261 gagacctacg tcgagcagca cgaggtggca gtggccagat actgcgacct ccctagcaaa
     1321 ctggggcaca agcttaattc cggagaaggg caagggcaag ggcaagggcc gggccgcggc
     1381 tacgcgtatc gatccatgtt cgaggcgcgc ctggtccagg gctccatcct caagaaggtg
     1441 ttggaggcac tcaaggacct catcaacgag gcctgctggg atattagctc cagcggtgta
     1501 aacctgcaga gcatggactc gtcccacgtc tctttggtgc agctcaccct gcggtctgag
     1561 ggcttcgaca cctaccgctg cgaccgcaac ctggccatgg gcgtgaacct caccagtatg
     1621 tccaaaatac taaaatgcgc cggcaatgaa gatatcatta cactaagggc cgaagataac
     1681 gcggatacct tggcgctagt atttgaagca ccaaaccagg agaaagtttc agactatgaa
     1741 atgaagttga tggatttaga tgttgaacaa cttggaattc cagaacagga gtacagctgt
     1801 gtagtaaaga tgccttctgg tgaatttgca cgtatatgcc gagatctcag ccatattgga
     1861 gatgctgttg taatttcctg tgcaaaagac ggagtgaaat tttctgcaag tggagaactt
     1921 ggaaatggaa acattaaatt gtcacagaca agtaatgtcg ataaagagga ggaagctgtt
     1981 accatagaga tgaatgaacc agttcaacta acttttgcac tgaggtacct gaacttcttt
     2041 acaaaagcca ctccactctc ttcaacggtg acactcagta tgtctgcaga tgtacccctt
     2101 gttgtagagt ataaaattgc ggatatggga cacttaaaat actacttggc tcccaagatc
     2161 gaggatgaag aaggatctta gggatccacc ggatctagat aactgatcat aatcagccat
     2221 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     2281 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     2341 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     2401 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     2461 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     2521 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     2581 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     2641 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     2701 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     2761 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     2821 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     2881 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2941 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3001 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     3061 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3121 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3181 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3241 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3301 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     3361 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     3421 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3481 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3541 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3601 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3661 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3721 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     3781 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     3841 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     3901 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     3961 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4021 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4081 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4141 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4201 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4261 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4321 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4381 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     4441 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     4501 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     4561 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     4621 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     4681 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4741 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4801 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4861 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4921 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4981 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5041 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5101 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5161 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5221 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5281 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5341 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5401 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5461 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     5521 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     5581 ccg
//
