LOCUS       sequence-199183-        2477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro expression of mCherry 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-mCherry
FEATURES             Location/Qualifiers
     source          1..2477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..81
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     RBS             234..256
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(287..306)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(288..306)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(334..922)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(414..433)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1033..1052)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1044..1859
                     /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(1117..1136)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(1877..1924)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1920..1938
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(join(1990..2477,1..226))
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYKA"
     primer_bind     complement(2271..2290)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 ccttggagcc gtacatgaac tgaggggaca ggatgtccca ggcgaagggc agggggccac
       61 ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc tcgccctcgc
      121 cctcgatctc gaactcgtgg ccgttcacgg agccctccat gtgcaccttg aagcgcatga
      181 actccttgat gatggccatg ttatcctcct cgcccttgct caccatatgt atatctcctt
      241 cttaaagtta aacaaaatta tttctagagg gaaaccgttg tggtctccct atagtgagtc
      301 gtattaattt cgcgggatcg cgttgctggc gtttttccat aggctccgcc cccctgacga
      361 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
      421 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      481 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
      541 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
      601 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
      661 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
      721 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
      781 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
      841 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
      901 gcgcagaaaa aaaggatctc aagaagatcc tttgatctca cgttgtgtct caaaatctct
      961 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
     1021 taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtct tgctcgaagc
     1081 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     1141 tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt
     1201 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
     1261 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     1321 atgcatggtt actcaccact gcgatccccg ggaaaacagc attccaggta ttagaagaat
     1381 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     1441 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
     1501 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     1561 ggcctgttga acaagtctgg aaagaaatgc ataagctttt gccattctca ccggattcag
     1621 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     1681 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     1741 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     1801 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     1861 cagaattggc tttcagcaaa aaacccctca agacccgttt agaggcccca aggggttatg
     1921 ctagttattg ctcagcggtg gcagcagcca actcagcttc ctttcgggct ttgttagcag
     1981 ccggtcgact taagccttgt acagctcgtc catgccgccg gtggagtggc ggccctcggc
     2041 gcgttcgtac tgttccacga tggtgtagtc ctcgttgtgg gaggtgatgt ccaacttgat
     2101 gttgacgttg taggcgccgg gcagctgcac gggcttcttg gccttgtagg tggtcttgac
     2161 ctcagcgtcg tagtggccgc cgtccttcag cttcagcctc tgcttgatct cgcccttcag
     2221 ggcgccgtcc tcggggtaca tccgctcgga ggaggcctcc cagcccatgg tcttcttctg
     2281 cattacgggg ccgtcggagg ggaagttggt gccgcgcagc ttcaccttgt agatgaactc
     2341 gccgtcctgc agggaggagt cctgggtcac ggtcaccacg ccgccgtcct cgaagttcat
     2401 cacgcgctcc cacttgaagc cctcggggaa ggacagcttc aagtagtcgg ggatgtcggc
     2461 ggggtgcttc acgtagg
//
