LOCUS       sequence-126639-        3299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  1 color weak reporter used in study.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3299)
  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 3299)
  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: pPhlF-BCD22-sfGFP-ssrA P64U26C39T11 v1-1 (632)
FEATURES             Location/Qualifiers
     source          1..3299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             158..871
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1072..1089)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1243..1831)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1323..1342)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2002..2793,2794..2862))
                     /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     2625..2644
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2863..2967)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3035..3053
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcattgctgt tcgattcgtt accaattgac atgatacgaa acgtaccgta tcgttaaggt
       61 tactagagaa gcgggcccaa gttcacttaa aaaggagatc aacaatgaaa gcaattttcg
      121 tactgaaaca tcttaatcat gcctaggaag ttttctaatg cgtaaaggag aagaactttt
      181 cactggtgtc gtccctattc tggtggaact ggatggtgat gtcaacggtc ataagttttc
      241 cgtgcgtggc gagggtgaag gtgacgcaac taatggtaaa ctgacgctga agttcatctg
      301 tactactggt aaactgccgg taccttggcc gactctggta acgacgctga cttatggtgt
      361 tcagtgcttt gctcgttatc cggaccatat gaagcagcat gacttcttca agtccgccat
      421 gccggaaggc tatgtgcagg aacgcacgat ttcctttaag gatgacggca cgtacaaaac
      481 gcgtgcggaa gtgaaatttg aaggcgatac cctggtaaac cgcattgagc tgaaaggcat
      541 tgactttaaa gaagatggca atatcctggg ccataagctg gaatacaatt ttaacagcca
      601 caatgtttac atcaccgccg ataaacaaaa aaatggcatt aaagcgaatt ttaaaattcg
      661 ccacaacgtg gaggatggca gcgtgcagct ggctgatcac taccagcaaa acactccaat
      721 cggtgatggt cctgttctgc tgccagacaa tcactatctg agcacgcaaa gcgttctgtc
      781 taaagatccg aacgagaaac gcgatcatat ggttctgctg gagttcgtaa ccgcagcggg
      841 catcacgcat ggtatggatg aactgtacaa agcagcaaac gacgaaaact acgctttagc
      901 agcttaatga actcaaagcc cgccgaaagg cgggcttttc tgtccttgag agtcgggcat
      961 tgtcttcgct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtg
     1021 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac
     1081 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1141 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1201 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1261 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1321 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1381 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1441 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1501 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1561 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1621 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1681 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1741 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     1801 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1861 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     1921 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1981 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2041 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2101 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgtgacccac gctcaccggc
     2161 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     2221 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     2281 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     2341 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttgcatgatc
     2401 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     2461 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     2521 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     2581 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca
     2641 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     2701 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     2761 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     2821 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     2881 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     2941 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     3001 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     3061 tgttcaagaa ttctggcgaa tcctctgacc agccagaaaa cgacctttct gtggtgaaac
     3121 cggatgctgc aattcagagc gccagcaagt gggggacagc agatgacctg accgccgcag
     3181 agtggatgtt tgacatggtg atgactatcg caccatcagc cagaaaaccg aattttgctg
     3241 ggtgggctaa cgatatccgc ctgatgcgtg aacgtgacgg acgtaacgaa gacaaccac
//
