LOCUS       sequence-474329-        4773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Endoplasmic Reticulum, Excitation: 399, Emission: 456.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4773)
  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 4773)
  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-ER-5
FEATURES             Location/Qualifiers
     source          1..4773
                     /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      603..612
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             609..1319
                     /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    1501..1622
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1538..1557)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1592..1611
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1629..2084)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1766..1787)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1978..1997
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2111..2215
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2217..2574
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2217..2408
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2240..2260)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2425..2560
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2609..3403
                     /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             2609..3403
                     /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(2663..2682)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3273..3292
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3591..3610)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3635..3682
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4011..4599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4500..4519
                     /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 gctagcgcca
      541 ccatgctgct atccgtgccg ttgctgctcg gcctcctcgg cctggccgtc gccgcaccgg
      601 tcgccaccat ggtgtctaag ggcgaagagc tgattaagga gaacatgcac atgaagctgt
      661 acatggaggg caccgtggac aaccatcact tcaagtgcac atccgagggc gaaggcaagc
      721 cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct ctccccttcg
      781 ccttcgacat cctggctact agcttcctct acggcagcaa gaccttcatc aaccacaccc
      841 agggcatccc cgacttcttc aagcagtcct tccctgaggg cttcacatgg gagagagtca
      901 ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct
      961 gcctcatcta caacgtcaag atcagagggg tgaacttcac atccaacggc cctgtgatgc
     1021 agaagaaaac actcggctgg gaggccttca ccgagacgct gtaccccgct gacggcggcc
     1081 tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg gagccatctg atcgcaaacg
     1141 ccaagaccac atatagatcc aagaaacccg ctaagaacct caagatgcct ggcgtctact
     1201 atgtggacta cagactggaa agaatcaagg aggccaacaa cgagacctac gtcgagcagc
     1261 acgaggtggc agtggccaga tactgcgacc tccctagcaa actggggcac aagcttaatt
     1321 ccggaaagga cgagctgtaa gaattctgca gtcgacggta ccgcgggccc gggatccacc
     1381 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
     1441 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     1501 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     1561 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     1621 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     1681 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     1741 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     1801 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     1861 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     1921 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     1981 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     2041 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     2101 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2161 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     2221 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2281 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2341 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2401 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2461 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     2521 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     2581 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     2641 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     2701 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     2761 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     2821 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     2881 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     2941 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3001 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3061 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3121 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     3181 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     3241 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     3301 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     3361 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     3421 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     3481 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     3541 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     3601 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     3661 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     3721 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     3781 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     3841 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     3901 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3961 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4021 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4081 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4141 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4201 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4261 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4321 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4381 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4441 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4501 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4561 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4621 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4681 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg
     4741 gtcattagtt catagcccat atatggagtt ccg
//
