LOCUS       sequence-199469-        2468 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro expression of tdTomato from the T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2468)
  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 2468)
  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-dTomato
FEATURES             Location/Qualifiers
     source          1..2468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(8..27)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     277..295
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     RBS             436..458
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(489..508)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(490..508)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(536..1124)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(616..635)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1235..1254)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1246..2061
                     /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"
     primer_bind     complement(1319..1338)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(2079..2126)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2122..2140
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(join(2195..2468,1..428))
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(2278..2301)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
ORIGIN
        1 cccatggtct tcttctgcat tacggggccg tcggggggga agttggtgcc gcgcatcttc
       61 accttgtaga tcagcgtgcc gtcctgcagg gaggagtcct gggtcacggt caccagaccg
      121 ccgtcctcga agttcatcac gcgctcccac ttgaagccct cggggaagga cagcttcttg
      181 taatcgggga tgtcggcggg gtgcttcacg tacgccttgg agccgtacat gaactggggg
      241 gacaggatgt cccaggcgaa gggcaggggg ccgcccttgg tcaccttcag cttggcggtc
      301 tgggtgccct cgtaggggcg gccctcgccc tcgccctcga tctcgaactc gtggccgttc
      361 atggagccct ccatgcgcac cttgaagcgc atgaactctt tgatgacctc ctcgcccttg
      421 ctcaccatat gtatatctcc ttcttaaagt taaacaaaat tatttctaga gggaaaccgt
      481 tgtggtctcc ctatagtgag tcgtattaat ttcgcgggat cgcgttgctg gcgtttttcc
      541 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
      601 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
      661 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
      721 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
      781 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
      841 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
      901 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
      961 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     1021 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1081 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1141 cacgttgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
     1201 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     1261 cgggaaacgt cttgctcgaa gccgcgatta aattccaaca tggatgctga tttatatggg
     1321 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg
     1381 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     1441 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     1501 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca
     1561 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     1621 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     1681 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     1741 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataagctt
     1801 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     1861 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     1921 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     1981 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     2041 atgctcgatg agtttttcta atcagaattg gctttcagca aaaaacccct caagacccgt
     2101 ttagaggccc caaggggtta tgctagttat tgctcagcgg tggcagcagc caactcagct
     2161 tcctttcggg ctttgttagc agccggtcga cttacttgta cagctcgtcc atgccgtaca
     2221 ggaacaggtg gtggcggccc tcggagcgct cgtactgttc cacgatggtg tagtcctcgt
     2281 tgtgggaggt gatgtccagc ttggtgtcca cgtagtagta gccgggcagt tgcacgggct
     2341 tcttggccat gtagatggtc ttgaactcca ccaggtagtg gccgccgtcc ttcagcttca
     2401 gggcctggtg gatctcgccc ttcagcacgc cgtcgcgggg gtacaggcgc tcggtggagg
     2461 cctcccag
//
