LOCUS       sequence-199484-        2477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro expression of mTagBFP2 from the T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2477)
  AUTHORS   Stark JC, Huang A, Nguyen PQ, Dubner RS, Hsu KJ, Ferrante TC, 
            Anderson M, Kanapskyte A, Mucha Q, Packett JS, Patel P, Patel R, Qaq
            D, Zondor T, Burke J, Martinez T, Miller-Berry A, Puppala A, 
            Reichert K, Schmid M, Brand L, Hill LR, Chellaswamy JF, Faheem N, 
            Fetherling S, Gong E, Gonzalzles EM, Granito T, Koritsaris J, Nguyen
            B, Ottman S, Palffy C, Patel A, Skweres S, Slaton A, Woods T, 
            Donghia N, Pardee K, Collins JJ, Jewett MC
  TITLE     BioBits Bright: A fluorescent synthetic biology education kit.
  JOURNAL   Sci Adv. 2018 Aug 1;4(8):eaat5107. doi: 10.1126/sciadv.aat5107. 
            eCollection 2018 Aug.
  PUBMED    30083609
REFERENCE   2  (bases 1 to 2477)
  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: "Sci Adv."; 
            date: "2018-08-1"; volume: "4(8):eaat5107. doi"; pages: " 
            10.1126/sciadv.aat5107. eCollection 2018 Aug"
COMMENT     Sequence Label: pJL1-mTagBFP2
FEATURES             Location/Qualifiers
     source          1..2477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     34..53
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     118..137
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      248..836
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     737..756
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     864..883
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        864..882
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             914..936
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             944..1654
                     /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"
     primer_bind     complement(1709..1727)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1723..1770
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(1788..2477,1..126))
                     /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="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
       61 atcagcatcc atgttggaat ttaatcgcgg cttcgagcaa gacgtttccc gttgaatatg
      121 gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga
      181 tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt gagatcaaag
      241 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
      301 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      361 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
      421 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      481 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      541 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      601 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
      661 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
      721 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
      781 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
      841 ccagcaacgc gatcccgcga aattaatacg actcactata gggagaccac aacggtttcc
      901 ctctagaaat aattttgttt aactttaaga aggagatata catatggtgt ctaagggcga
      961 agagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tggacaacca
     1021 tcacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag
     1081 aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctactagctt
     1141 cctctacggc agcaagacct tcatcaacca cacccagggc atccccgact tcttcaagca
     1201 gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct
     1261 gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag
     1321 aggggtgaac ttcacatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc
     1381 cttcaccgag acgctgtacc ccgctgacgg cggcctggaa ggcagaaacg acatggccct
     1441 gaagctcgtg ggcgggagcc atctgatcgc aaacgccaag accacatata gatccaagaa
     1501 acccgctaag aacctcaaga tgcctggcgt ctactatgtg gactacagac tggaaagaat
     1561 caaggaggcc aacaacgaga cctacgtcga gcagcacgag gtggcagtgg ccagatactg
     1621 cgacctccct agcaaactgg ggcacaagct taattaagtc gaccggctgc taacaaagcc
     1681 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     1741 gcctctaaac gggtcttgag gggttttttg ctgaaagcca attctgatta gaaaaactca
     1801 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     1861 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     1921 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     1981 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     2041 aatggcaaaa gcttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     2101 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     2161 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     2221 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     2281 tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     2341 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     2401 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     2461 tcgggcttcc catacaa
//
