LOCUS       sequence-199468-        2441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro expression of mRFP1 from the T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2441)
  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 2441)
  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-mRFP1
FEATURES             Location/Qualifiers
     source          1..2441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     221..240
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     348..367
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        348..366
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             398..420
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             428..1105
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1157..1175)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1171..1218
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1236..2051)
                     /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     1959..1978
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2043..2062
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(2173..2441,1..320)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
       61 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
      121 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
      181 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
      241 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
      301 gggggcggag cctatggaaa aacgccagca acgcgatccc gcgaaattaa tacgactcac
      361 tatagggaga ccacaacggt ttccctctag aaataatttt gtttaacttt aagaaggaga
      421 tatacatatg gcttcctccg aagacgttat caaagagttc atgcgtttca aagttcgtat
      481 ggaaggttcc gttaacggtc acgagttcga aatcgaaggt gaaggtgaag gtcgtccgta
      541 cgaaggtacc cagaccgcta aactgaaagt taccaaaggt ggtccgctgc cgttcgcttg
      601 ggacatcctg tccccgcagt tccagtacgg ttccaaagct tacgttaaac acccggctga
      661 catcccggac tacctgaaac tgtccttccc ggaaggtttc aaatgggaac gtgttatgaa
      721 cttcgaagac ggtggtgttg ttaccgttac ccaggactcc tccctgcaag acggtgagtt
      781 catctacaaa gttaaactgc gtggtaccaa cttcccgtcc gacggtccgg ttatgcagaa
      841 aaaaaccatg ggttgggaag cttccaccga acgtatgtac ccggaagacg gtgctctgaa
      901 aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt cactacgacg ctgaagttaa
      961 aaccacctac atggctaaaa aaccggttca gctgccgggt gcttacaaaa ccgacatcaa
     1021 actggacatc acctcccaca acgaagacta caccatcgtt gaacagtacg aacgtgctga
     1081 aggtcgtcac tccaccggtg cttaagtcga ccggctgcta acaaagcccg aaaggaagct
     1141 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     1201 gtcttgaggg gttttttgct gaaagccaat tctgattaga aaaactcatc gagcatcaaa
     1261 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     1321 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     1381 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     1441 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc
     1501 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     1561 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     1621 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     1681 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     1741 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     1801 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     1861 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     1921 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     1981 tataaatcag catccatgtt ggaatttaat cgcggcttcg agcaagacgt ttcccgttga
     2041 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     2101 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtgagat
     2161 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2221 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2281 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2341 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2401 accagtggct gctgccagtg gcgataagtc gtgtcttacc g
//
