LOCUS       sequence-154499-        3366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  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 3366)
  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 3366)
  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
FEATURES             Location/Qualifiers
     source          1..3366
                     /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..1467
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     protein_bind    complement(1475..1570)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1607..1626)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1608..1626)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1631..1647)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1631..1647)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1749..1768)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1760..2569
                     /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(1827..1846)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2716..3304
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3205..3224
                     /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 cagtctctag agctcgaatt 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 taattaaact agtgcaactt tattatacaa agttggcatt
     1501 ataaaaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa
     1561 atcattattt ggagctccat ggtagcgtta acgcggccgc gatatcccct atagtgagtc
     1621 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     1681 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     1741 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     1801 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     1861 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     1921 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     1981 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     2041 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     2101 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     2161 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     2221 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     2281 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     2341 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     2401 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     2461 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     2521 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     2581 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     2641 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     2701 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2761 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2821 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     2881 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2941 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3001 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3061 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc
     3121 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3181 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3241 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3301 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3361 catgtt
//
