LOCUS       sequence-126627-        4450 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid for building 5n2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4450)
  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 4450)
  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: pBetI-BCD7-qacR-ssrA P69U18C63T16 v50-2 (568)
FEATURES             Location/Qualifiers
     source          1..4450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      972..1043
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1059..1086
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1617..2567)
                     /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(3329..3423)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(3454..4245,4246..4314))
                     /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     4077..4096
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gacgtcggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
       61 ttccagtcgg gaaacctgaa gacaactcgg cattgctgtt cgattcgtta ccaattgaca
      121 attgattgga cgttcaatat aatgctagct actagagaag cgggcccaag ttcacttaaa
      181 aaggagatca acaatgaaag caattttcgt actgaaacat cttaatcatg ctggggaggg
      241 tttctaatga acctgaaaga taaaattctg ggcgttgcca aagaactgtt tatcaaaaat
      301 ggctataacg caaccaccac cggtgaaatt gttaaactga gcgaaagcag caaaggcaat
      361 ctgtattatc actttaaaac caaagagaac ctgtttctgg aaatcctgaa catcgaagaa
      421 agcaaatggc aagagcagtg gaaaaaagaa caaatcaaat gcaaaaccaa ccgcgagaaa
      481 ttctatctgt ataatgaact gagcctgacc accgaatatt actatccgct gcagaatgcc
      541 atcatcgagt tttataccga gtactataaa accaacagca tcaacgagaa aatgaacaaa
      601 ctggaaaaca aatacatcga tgcctaccac gtgatcttta aagaaggtaa tctgaacggc
      661 gaatggtgca ttaatgatgt taatgccgtg agcaaaattg cagcaaatgc cgttaatggc
      721 attgttacct ttacccatga gcagaatatc aacgaacgca ttaaactgat gaacaaattc
      781 agccagatct ttctgaatgg cctgagcaaa gcagcaaacg acgaaaacta cgctttagca
      841 gcttaatgaa ctgttatgag tcaggaaaaa aggcgacaga gtaatctgtc gccttttttc
      901 tttgcttgct ttccttgaga gtcgaacctt gtcttctaag gatccaaact cgagtaagga
      961 tctccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     1021 gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt
     1081 tctgcgttta tacctagggt acgggttttg ctgcccgcaa acgggctgtt ctggtgttgc
     1141 tagtttgtta tcagaatcgc agatccggct tcagccggtt tgccggctga aagcgctatt
     1201 tcttccagaa ttgccatgat tttttcccca cgggaggcgt cactggctcc cgtgttgtcg
     1261 gcagctttga ttcgataagc agcatcgcct gtttcaggct gtctatgtgt gactgttgag
     1321 ctgtaacaag ttgtctcagg tgttcaattt catgttctag ttgctttgtt ttactggttt
     1381 cacctgttct attaggtgtt acatgctgtt catctgttac attgtcgatc tgttcatggt
     1441 gaacagcttt gaatgcacca aaaactcgta aaagctctga tgtatctatc ttttttacac
     1501 cgttttcatc tgtgcatatg gacagttttc cctttgatat gtaacggtga acagttgttc
     1561 tacttttgtt tgttagtctt gatgcttcac tgatagatac aagagccata agaacctcag
     1621 atccttccgt atttagccag tatgttctct agtgtggttc gttgtttttg cgtgagccat
     1681 gagaacgaac cattgagatc atacttactt tgcatgtcac tcaaaaattt tgcctcaaaa
     1741 ctggtgagct gaatttttgc agttaaagca tcgtgtagtg tttttcttag tccgttatgt
     1801 aggtaggaat ctgatgtaat ggttgttggt attttgtcac cattcatttt tatctggttg
     1861 ttctcaagtt cggttacgag atccatttgt ctatctagtt caacttggaa aatcaacgta
     1921 tcagtcgggc ggcctcgctt atcaaccacc aatttcatat tgctgtaagt gtttaaatct
     1981 ttacttattg gtttcaaaac ccattggtta agccttttaa actcatggta gttattttca
     2041 agcattaaca tgaacttaaa ttcatcaagg ctaatctcta tatttgcctt gtgagttttc
     2101 ttttgtgtta gttcttttaa taaccactca taaatcctca tagagtattt gttttcaaaa
     2161 gacttaacat gttccagatt atattttatg aattttttta actggaaaag ataaggcaat
     2221 atctcttcac taaaaactaa ttctaatttt tcgcttgaga acttggcata gtttgtccac
     2281 tggaaaatct caaagccttt aaccaaagga ttcctgattt ccacagttct cgtcatcagc
     2341 tctctggttg ctttagctaa tacaccataa gcattttccc tactgatgtt catcatctga
     2401 gcgtattggt tataagtgaa cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg
     2461 gggttgagta gtgccacaca gcataaaatt agcttggttt catgctccgt taagtcatag
     2521 cgactaatcg ctagttcatt tgctttgaaa acaactaatt cagacataca tctcaattgg
     2581 tctaggtgat tttaatcact ataccaattg agatgggcta gtcaatgata attactagtc
     2641 cttttcccgg gtgatctggg tatctgtaaa ttctgctaga cctttgctgg aaaacttgta
     2701 aattctgcta gaccctctgt aaattccgct agacctttgt gtgttttttt tgtttatatt
     2761 caagtggtta taatttatag aataaagaaa gaataaaaaa agataaaaag aatagatccc
     2821 agccctgtgt ataactcact actttagtca gttccgcagt attacaaaag gatgtcgcaa
     2881 acgctgtttg ctcctctaca aaacagacct taaaacccta aaggcttaag tagcaccctc
     2941 gcaagctcgg gcaaatcgct gaatattcct tttgtctccg accatcaggc acctgagtcg
     3001 ctgtcttttt cgtgacattc agttcgctgc gctcacggct ctggcagtga atgggggtaa
     3061 atggcactac aggcgccttt tatggattca tgcaaggaaa ctacccataa tacaagaaaa
     3121 gcccgtcacg ggcttctcag ggcgttttat ggcgggtctg ctatgtggtg ctatctgact
     3181 ttttgctgtt cagcagttcc tgccctctga ttttccagtc tgaccacttc ggattatccc
     3241 gtgacaggtc attcagactg gctaatgcac ccagtaaggc agcggtatca tcaacaggct
     3301 tacccgtctt actgtcccta gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac
     3361 cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga
     3421 gtccaagcga gctctcgaac cccagagtcc cgcttaccaa tgcttaatca gtgaggcacc
     3481 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3541 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgtgaccc
     3601 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3661 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3721 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3781 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3841 agttgcatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3901 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3961 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4021 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa
     4081 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4141 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4201 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4261 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatgcgaaa
     4321 cgatcctcat cctgtctctt gatcagatca tgatcccctg cgccatcaga tccttggcgg
     4381 caagaaagcc atccagttta ctttgcaggg cttcccaacc ttaccagagg gcgccccagc
     4441 tggcaattcc
//
