LOCUS       sequence-126641-        3345 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 3345)
  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 3345)
  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: pSrpR-BCD20-Cerulean-ssrA P65U23C89T19 v50-2a ...
FEATURES             Location/Qualifiers
     source          1..3345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             177..890
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             177..890
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(207..235)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     830..850
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1118..1135)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1289..1877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1369..1388)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2048..2839,2840..2908))
                     /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     2671..2690
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2909..3013)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3081..3099
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcattgctgt tcgattcgtt accaattgac agctagctca gtcctaggta tatacataca
       61 tgcttgtttg tttgtaaact actagagaag cgggcccaag ttcacttaaa aaggagatca
      121 acaatgaaag caattttcgt actgaaacat cttaatcatg ctgaggaaag tttctaatga
      181 gtaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg
      241 ttaatgggca caaattttct gtcagtggag agggtgaagg tgatgcaaca tacggaaaac
      301 ttacccttaa atttatttgc actactggaa aactacctgt tccatggcca acacttgtca
      361 ctactttgac ttggggtgtt caatgctttg ctagataccc agatcatatg aaacagcatg
      421 actttttcaa gagtgccatg cccgaaggtt atgtacagga aagaactata tttttcaaag
      481 atgacgggaa ctacaagaca cgtgctgaag tcaagtttga aggtgatacc cttgttaata
      541 gaatcgagtt aaaaggtatt gattttaaag aagatggaaa cattcttgga cacaaattgg
      601 aatacaacgc tatttcagat aatgtataca tcactgcaga caaacaaaag aatggaatca
      661 aagctaattt caaaattaga cacaacattg aagatggaag cgttcaacta gcagaccatt
      721 atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac cattacctgt
      781 ccacacaatc tgccctttcg aaagatccca acgaaaagag agatcacatg gtccttcttg
      841 agtttgtaac agctgctggg attacactag gcatggatga actatacaaa gcagcaaacg
      901 acgaaaacta cgctttagca gcttaatgaa ctctcggtac caaattccag aaaagagacg
      961 ctttcgagcg tcttttttcg ttttggtccc cttgagagtc gatgtttgtc ttcgctcctt
     1021 ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtgttct ttatcatgca
     1081 actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct
     1141 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     1201 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     1261 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1321 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1381 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1441 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1501 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1561 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1621 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1681 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1741 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1801 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1861 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1921 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1981 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2041 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2101 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     2161 ctggccccag tgctgcaatg ataccgcgtg acccacgctc accggctcca gatttatcag
     2221 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     2281 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     2341 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     2401 cttcattcag ctccggttcc caacgatcaa ggcgagttgc atgatccccc atgttgtgca
     2461 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     2521 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     2581 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     2641 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
     2701 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     2761 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     2821 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     2881 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     2941 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3001 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     3061 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtgtt caagaattct
     3121 ggcgaatcct ctgaccagcc agaaaacgac ctttctgtgg tgaaaccgga tgctgcaatt
     3181 cagagcgcca gcaagtgggg gacagcagat gacctgaccg ccgcagagtg gatgtttgac
     3241 atggtgatga ctatcgcacc atcagccaga aaaccgaatt ttgctgggtg ggctaacgat
     3301 atccgcctga tgcgtgaacg tgacggacgt aacgaagaca aggca
//
