LOCUS       sequence-126618-        6231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BetI/PhlF/SrpR BCD7 3-node v41-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6231)
  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 6231)
  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: 3n1 (572)
FEATURES             Location/Qualifiers
     source          1..6231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2819..2890
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2906..2933
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3464..4414)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(5176..5270)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5301..6095)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5412..5431)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6022..6041
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
       61 ttccagtcgg gaaacctggt ctcaaagccc acgcattgct gttcgattcg ttaccaattg
      121 acatgatacg aaacgtaccg tatcgttaag gttactagag aagcgggccc aagttcactt
      181 aaaaaggaga tcaacaatga aagcaatttt cgtactgaaa catcttaatc atgctgggga
      241 gggtttctaa tggcacgtaa aaccgcagca gaagcagaag aaacccgtca gcgtattatt
      301 gatgcagcac tggaagtttt tgttgcacag ggtgttagtg atgcaaccct ggatcagatt
      361 gcacgtaaag ccggtgttac ccgtggtgca gtttattggc attttaatgg taaactggaa
      421 gttctgcagg cagttctggc aagccgtcag catccgctgg aactggattt tacaccggat
      481 ctgggtattg aacgtagctg ggaagcagtt gttgttgcaa tgctggatgc agttcatagt
      541 ccgcagagca aacagtttag cgaaattctg atttatcagg gtctggatga aagcggtctg
      601 attcataatc gtatggttca ggcaagcgat cgttttctgc agtatattca tcaggttctg
      661 cgtcatgcag ttacccaggg tgaactgccg attaatctgg atctgcagac cagcattggt
      721 gtttttaaag gtctgattac cggtctgctg tatgaaggtc tgcgtagcaa agatcagcag
      781 gcacagatta tcaaagttgc actgggtagc ttttgggcac tgctgcgtga accgcctcgt
      841 tttctgctgt gtgaagaagc acagattaaa caggtgaaat ccttcgaagc agcaaacgac
      901 gaaaactacg ctttagcagc ttaatgaact ttcagccaaa aaacttaaga ccgccggtct
      961 tgtccactac cttgcagtaa tgcggtggac aggatcggcg gttttctttt ctcttctcaa
     1021 ccttgagagt cgggcagcat tgctgttcga ttcgttacca attgacagct agctcagtcc
     1081 taggtatata catacatgct tgtttgtttg taaactacta gagaagcggg cccaagttca
     1141 cttaaaaagg agatcaacaa tgaaagcaat tttcgtactg aaacatctta atcatgctgg
     1201 ggagggtttc taatgccgaa actgggtatg cagagcattc gtcgtcgtca gctgattgat
     1261 gcaaccctgg aagcaattaa tgaagttggt atgcatgatg caaccattgc acagattgca
     1321 cgtcgtgccg gtgttagcac cggtattatt agccattatt tccgcgataa aaacggtctg
     1381 ctggaagcaa ccatgcgtga tattaccagc cagctgcgtg atgcagttct gaatcgtctg
     1441 catgcactgc cgcagggtag cgcagaacag cgtctgcagg caattgttgg tggtaatttt
     1501 gatgaaaccc aggttagcag cgcagcaatg aaagcatggc tggcattttg ggcaagcagc
     1561 atgcatcagc cgatgctgta tcgtctgcag caggttagca gtcgtcgtct gctgagcaat
     1621 ctggttagcg aatttcgtcg tgaactgcct cgtgaacagg cacaagaggc aggttatggt
     1681 ctggcagcac tgattgatgg tctgtggctg cgtgcagcac tgagcggtaa accgctggat
     1741 aaaacccgtg caaatagcct gacccgtcat tttatcaccc agcatctgcc gaccgatgca
     1801 gcaaacgacg aaaactacgc tttagcagct taatgaacta acgcatgaga aagcccccgg
     1861 aagatcacct tccgggggct tttttattgc gcccttgaga gtcgatgtgc attgctgttc
     1921 gattcgttac caattgacaa ttgattggac gttcaatata atgctagcta ctagagaagc
     1981 gggcccaagt tcacttaaaa aggagatcaa caatgaaagc aattttcgta ctgaaacatc
     2041 ttaatcatgc tggggagggt ttctaatggc acgtaccccg agccgtagca gcattggtag
     2101 cctgcgtagt ccgcataccc ataaagcaat tctgaccagc accattgaaa tcctgaaaga
     2161 atgtggttat agcggtctga gcattgaaag cgttgcacgt cgtgccggtg caagcaaacc
     2221 gaccatttat cgttggtgga ccaataaagc agcactgatt gccgaagtgt atgaaaatga
     2281 aagcgaacag gtgcgtaaat ttccggatct gggtagcttt aaagccgatc tggattttct
     2341 gctgcgtaat ctgtggaaag tttggcgtga aaccatttgt ggtgaagcat ttcgttgtgt
     2401 tattgcagaa gcacagctgg accctgcaac cctgacccag ctgaaagatc agtttatgga
     2461 acgtcgtcgt gagatgccga aaaaactggt tgaaaatgcc attagcaatg gtgaactgcc
     2521 gaaagatacc aatcgtgaac tgctgctgga tatgattttt ggtttttgtt ggtatcgcct
     2581 gctgaccgaa cagctgaccg ttgaacagga tattgaagaa tttaccttcc tgctgattaa
     2641 tggtgtttgt ccgggtacac agcgtgcagc aaacgacgaa aactacgctt tagcagctta
     2701 atgaacttcc ggcaattaaa aaagcggcta accacgccgc tttttttacg tctgcacctt
     2761 gagagtcgct cggtcgtgag acctaaggat ccaaactcga gtaaggatct ccaggcatca
     2821 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     2881 gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct gcgtttatac
     2941 ctagggtacg ggttttgctg cccgcaaacg ggctgttctg gtgttgctag tttgttatca
     3001 gaatcgcaga tccggcttca gccggtttgc cggctgaaag cgctatttct tccagaattg
     3061 ccatgatttt ttccccacgg gaggcgtcac tggctcccgt gttgtcggca gctttgattc
     3121 gataagcagc atcgcctgtt tcaggctgtc tatgtgtgac tgttgagctg taacaagttg
     3181 tctcaggtgt tcaatttcat gttctagttg ctttgtttta ctggtttcac ctgttctatt
     3241 aggtgttaca tgctgttcat ctgttacatt gtcgatctgt tcatggtgaa cagctttgaa
     3301 tgcaccaaaa actcgtaaaa gctctgatgt atctatcttt tttacaccgt tttcatctgt
     3361 gcatatggac agttttccct ttgatatgta acggtgaaca gttgttctac ttttgtttgt
     3421 tagtcttgat gcttcactga tagatacaag agccataaga acctcagatc cttccgtatt
     3481 tagccagtat gttctctagt gtggttcgtt gtttttgcgt gagccatgag aacgaaccat
     3541 tgagatcata cttactttgc atgtcactca aaaattttgc ctcaaaactg gtgagctgaa
     3601 tttttgcagt taaagcatcg tgtagtgttt ttcttagtcc gttatgtagg taggaatctg
     3661 atgtaatggt tgttggtatt ttgtcaccat tcatttttat ctggttgttc tcaagttcgg
     3721 ttacgagatc catttgtcta tctagttcaa cttggaaaat caacgtatca gtcgggcggc
     3781 ctcgcttatc aaccaccaat ttcatattgc tgtaagtgtt taaatcttta cttattggtt
     3841 tcaaaaccca ttggttaagc cttttaaact catggtagtt attttcaagc attaacatga
     3901 acttaaattc atcaaggcta atctctatat ttgccttgtg agttttcttt tgtgttagtt
     3961 cttttaataa ccactcataa atcctcatag agtatttgtt ttcaaaagac ttaacatgtt
     4021 ccagattata ttttatgaat ttttttaact ggaaaagata aggcaatatc tcttcactaa
     4081 aaactaattc taatttttcg cttgagaact tggcatagtt tgtccactgg aaaatctcaa
     4141 agcctttaac caaaggattc ctgatttcca cagttctcgt catcagctct ctggttgctt
     4201 tagctaatac accataagca ttttccctac tgatgttcat catctgagcg tattggttat
     4261 aagtgaacga taccgtccgt tctttccttg tagggttttc aatcgtgggg ttgagtagtg
     4321 ccacacagca taaaattagc ttggtttcat gctccgttaa gtcatagcga ctaatcgcta
     4381 gttcatttgc tttgaaaaca actaattcag acatacatct caattggtct aggtgatttt
     4441 aatcactata ccaattgaga tgggctagtc aatgataatt actagtcctt ttcccgggtg
     4501 atctgggtat ctgtaaattc tgctagacct ttgctggaaa acttgtaaat tctgctagac
     4561 cctctgtaaa ttccgctaga cctttgtgtg ttttttttgt ttatattcaa gtggttataa
     4621 tttatagaat aaagaaagaa taaaaaaaga taaaaagaat agatcccagc cctgtgtata
     4681 actcactact ttagtcagtt ccgcagtatt acaaaaggat gtcgcaaacg ctgtttgctc
     4741 ctctacaaaa cagaccttaa aaccctaaag gcttaagtag caccctcgca agctcgggca
     4801 aatcgctgaa tattcctttt gtctccgacc atcaggcacc tgagtcgctg tctttttcgt
     4861 gacattcagt tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg gcactacagg
     4921 cgccttttat ggattcatgc aaggaaacta cccataatac aagaaaagcc cgtcacgggc
     4981 ttctcagggc gttttatggc gggtctgcta tgtggtgcta tctgactttt tgctgttcag
     5041 cagttcctgc cctctgattt tccagtctga ccacttcgga ttatcccgtg acaggtcatt
     5101 cagactggct aatgcaccca gtaaggcagc ggtatcatca acaggcttac ccgtcttact
     5161 gtccctagtg cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa
     5221 caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct
     5281 ctcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc
     5341 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca
     5401 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc
     5461 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag
     5521 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg
     5581 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca
     5641 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat
     5701 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact
     5761 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc
     5821 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc
     5881 gtggccagcc acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg
     5941 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca
     6001 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc
     6061 ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct
     6121 tgatcagatc atgatcccct gcgccatcag atccttggcg gcaagaaagc catccagttt
     6181 actttgcagg gcttcccaac cttaccagag ggcgccccag ctggcaattc c
//
