LOCUS       sequence-126642-        3314 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for building 3-color reporter plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3314)
  AUTHORS   Niederholtmeyer H, Sun ZZ, Hori Y, Yeung E, Verpoorte A, Murray RM, 
            Maerkl SJ
  TITLE     Rapid cell-free forward engineering of novel genetic ring 
            oscillators.
  JOURNAL   Elife. 2015 Oct 2;4. doi: 10.7554/eLife.09771.
  PUBMED    26430766
REFERENCE   2  (bases 1 to 3314)
  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: "Elife."; 
            date: "2015-10-2"; volume: "4. doi"; pages: " 10.7554/eLife.09771"
COMMENT     Sequence Label: pTetR-BCD20-mCherry-ssrA P66U23C90T20 v40-3a (...
FEATURES             Location/Qualifiers
     source          1..3314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             158..865
                     /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
                     EGRHSTGGMDELYK"
     primer_bind     complement(303..321)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(1087..1104)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1258..1846)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1338..1357)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2017..2808,2809..2877))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHATRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2640..2659
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2878..2982)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3050..3068
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcattgctgt tctcagtgat agagattgac atccctatca gtgatagata taatgagcac
       61 tactagagaa gcgggcccaa gttcacttaa aaaggagatc aacaatgaaa gcaattttcg
      121 tactgaaaca tcttaatcat gctgaggaaa gtttctaatg gtgagcaagg gcgaggagga
      181 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
      241 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
      301 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
      361 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
      421 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
      481 cgtggtgacc gtgacccagg actcctcctt gcaggacggc gagttcatct acaaggtgaa
      541 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaaa ccatgggctg
      601 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
      661 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
      721 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
      781 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
      841 cggcggcatg gacgagctgt acaaggcagc aaacgacgaa aactacgctt tagcagctta
      901 atgaactgac gaacaataag gcctccctaa cggggggcct tttttattga taacaaaacc
      961 ttgagagtcg ctcgttgtct tcgctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     1021 gcacttatga ctgtgttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc
     1081 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1141 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1201 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1261 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1321 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1381 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1441 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1501 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1561 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1621 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1681 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1741 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1801 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1861 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1921 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     1981 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2041 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2101 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgtga
     2161 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2221 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2281 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2341 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2401 gcgagttgca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     2461 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     2521 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     2581 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga
     2641 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     2701 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     2761 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     2821 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     2881 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     2941 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3001 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     3061 cacgaggccc tttcgtgttc aagaattctg gcgaatcctc tgaccagcca gaaaacgacc
     3121 tttctgtggt gaaaccggat gctgcaattc agagcgccag caagtggggg acagcagatg
     3181 acctgaccgc cgcagagtgg atgtttgaca tggtgatgac tatcgcacca tcagccagaa
     3241 aaccgaattt tgctgggtgg gctaacgata tccgcctgat gcgtgaacgt gacggacgta
     3301 acgaagacaa atgt
//
