LOCUS       sequence-199486-        2483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro expression of eBFP2 from the T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2483)
  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 2483)
  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-eBFP2
FEATURES             Location/Qualifiers
     source          1..2483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     436..457
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     RBS             510..532
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(563..582)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(564..582)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(610..1198)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(690..709)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1309..1328)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1320..2135
                     /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(1393..1412)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(2153..2200)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2196..2214
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(join(2266..2483,1..496,497..499,500..502))
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
ORIGIN
        1 tcttgaagtt caccttgatg ccgttcttct gcttgacggc catgatatag atgttgtggc
       61 tgttgaagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgacgc
      121 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct
      181 tgtaggtgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg
      241 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcgggcgaag cactgcacgc
      301 cgtggctcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga
      361 tgaacttcag ggtcagcttg ccgttggtgg catcgccctc gccctcgccc ctcacgctga
      421 acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca
      481 gctcctcgcc cttgctcacc atatgtatat ctccttctta aagttaaaca aaattatttc
      541 tagagggaaa ccgttgtggt ctccctatag tgagtcgtat taatttcgcg ggatcgcgtt
      601 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      661 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
      721 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
      781 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
      841 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
      901 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
      961 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1021 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1081 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1141 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1201 agatcctttg atctcacgtt gtgtctcaaa atctctgatg ttacattgca caagataaaa
     1261 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     1321 tgagccatat tcaacgggaa acgtcttgct cgaagccgcg attaaattcc aacatggatg
     1381 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     1441 atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     1501 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     1561 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     1621 tccccgggaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     1681 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     1741 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     1801 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     1861 aaatgcataa gcttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     1921 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     1981 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     2041 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     2101 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggctttc agcaaaaaac
     2161 ccctcaagac ccgtttagag gccccaaggg gttatgctag ttattgctca gcggtggcag
     2221 cagccaactc agcttccttt cgggctttgt tagcagccgg tcgacttact tgtacagctc
     2281 gtccatgccg agagtgatcc cggcggcggt gcggaactcc agcaggacca tgtgatcgcg
     2341 cttctcgttg gggtctttgc tcagcacgga ctgggtgctc aggtagtggc tgtcgggcag
     2401 cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct
     2461 gccgtcctcc acgttgtggc gga
//
