LOCUS       sequence-154502-        3441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Membrane targeted TagBFP (blue) fluorophore with stop codon for 
            C-terminal fusions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3441)
  AUTHORS   Don EK, Formella I, Badrock AP, Hall TE, Morsch M, Hortle E, Hogan 
            A, Chow S, Gwee SS, Stoddart JJ, Nicholson G, Chung R, Cole NJ
  TITLE     A Tol2 Gateway-Compatible Toolbox for the Study of the Nervous 
            System and Neurodegenerative Disease.
  JOURNAL   Zebrafish. 2016 Sep 15.
  PUBMED    27631880
REFERENCE   2  (bases 1 to 3441)
  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: "Zebrafish. 
            2016 Sep 15."
COMMENT     Sequence Label: p3E-mTagBFP-CAAX
FEATURES             Location/Qualifiers
     source          1..3441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(625..748)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             766..1464
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     protein_bind    complement(1550..1645)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1682..1701)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1683..1701)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1706..1722)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1706..1722)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1824..1843)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1835..2644
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1902..1921)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2791..3379
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3280..3299
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggccctg cagctctaga gctcgaattt ctacaggtca
      601 ctaataccat ctaagtagtt ggttcatagt gactgcatat gttgtgtttt acagtattat
      661 gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt
      721 tctcgttcaa ctttcttgta caaagtggct ggatccttcg ccaccatgag cgagctgatt
      781 aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tggacaacca tcacttcaag
      841 tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg
      901 gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctactagctt cctctacggc
      961 agcaagacct tcatcaacca cacccagggc atccccgact tcttcaagca gtccttccct
     1021 gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc
     1081 caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac
     1141 ttcacatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc cttcaccgag
     1201 acgctgtacc ccgctgacgg cggcctggaa ggcagaaacg acatggccct gaagctcgtg
     1261 ggcgggagcc atctgatcgc aaacatcaag accacatata gatccaagaa acccgctaag
     1321 aacctcaaga tgcctggcgt ctactatgtg gactacagac tggaaagaat caaggaggcc
     1381 aacaacgaga cctacgtcga gcagcacgag gtggcagtgg ccagatactg cgacctccct
     1441 agcaaactgg ggcacaagct taatggagga ggaagatcta agctgaaccc tcctgatgag
     1501 agtggccccg gctgcatgag ctgcaagtgt gtgctctcct gaactagtgc aactttatta
     1561 tacaaagttg gcattataaa aaagcattgc ttatcaattt gttgcaacga acaggtcact
     1621 atcagtcaaa ataaaatcat tatttggagc tccatggtag cgttaacgcg gccgcgatat
     1681 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     1741 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     1801 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     1861 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     1921 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     1981 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2041 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2101 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     2161 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     2221 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     2281 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     2341 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     2401 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     2461 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     2521 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     2581 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     2641 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     2701 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     2761 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2821 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2881 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     2941 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3001 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3061 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3121 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3181 gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3241 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3301 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3361 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3421 ctggcctttt gctcacatgt t
//
