LOCUS       sequence-199473-        2474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro expression of mTFP1 from the T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2474)
  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 2474)
  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-mTFP1
FEATURES             Location/Qualifiers
     source          1..2474
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             576..598
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(629..648)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(630..648)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(676..1264)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(756..775)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1375..1394)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1386..2201
                     /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(1459..1478)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(2219..2266)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2262..2280
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(join(2332..2474,1..568))
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
ORIGIN
        1 tcttggccct gtagatggtc ttgaagtcaa cgcggtggtg gccgccgccc tccagcagca
       61 gcttgtgctt gacgtcgccc ttcagcacgc cgtcgcgcac gtacatcctc tcggtggagg
      121 cgtcccagcc ggtggtcttc ttctgcatca cggggccgtt gggggggaag ttctcgccct
      181 tgaggtgtat ctcgtagatg aaggagtcct cctccatgga gatgtcggac ttcaccttca
      241 cgatgccctt gtcctcgaag gtcatggtgc gctcccaaga gtagccctcg gggaaggact
      301 gcttgaagta gttggggatg tcgtcggggt acttggtgaa ggccctgttg ccgtaggcga
      361 acgcggtggt cagaatgtcg taggagaagg gcaggggggc tccctccttc acctccaggt
      421 tgatggtgtt ggtgccgtcg tagggcttgc cctcgccctc gccctcgatc acgaaggcgt
      481 ggccattcac gttgccctcc atcttcagct tgatcttcat gtcgggcttg attacgccca
      541 ttgtggtctc ctcgcccttg ctcaccatat gtatatctcc ttcttaaagt taaacaaaat
      601 tatttctaga gggaaaccgt tgtggtctcc ctatagtgag tcgtattaat ttcgcgggat
      661 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      721 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      781 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      841 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      901 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      961 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1021 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1081 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1141 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1201 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1261 tcaagaagat cctttgatct cacgttgtgt ctcaaaatct ctgatgttac attgcacaag
     1321 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     1381 gtgttatgag ccatattcaa cgggaaacgt cttgctcgaa gccgcgatta aattccaaca
     1441 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     1501 caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     1561 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     1621 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     1681 ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     1741 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     1801 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     1861 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     1921 ggaaagaaat gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt
     1981 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     2041 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     2101 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     2161 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gctttcagca
     2221 aaaaacccct caagacccgt ttagaggccc caaggggtta tgctagttat tgctcagcgg
     2281 tggcagcagc caactcagct tcctttcggg ctttgttagc agccggtcga cttacttgta
     2341 cagctcgtcc atgccgtcgg tggagttgcg ggccacggcg ctctcgtaaa cggtcacctt
     2401 gttgtagtcc ttgtcgtggt tcaggatctc gatgcggtgg tccacaaagt gatagtcggg
     2461 cagcttcacc gcct
//
