LOCUS       sequence-126630-        4483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid for building 3n1, 5n2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4483)
  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 4483)
  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-BCD7-betI-ssrA P65U18C64T14 v50-2 (567)
FEATURES             Location/Qualifiers
     source          1..4483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1005..1076
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1092..1119
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1650..2600)
                     /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(3362..3456)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(3487..4278,4279..4347))
                     /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     4110..4129
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gacgtcggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
       61 ttccagtcgg gaaacctgaa gacaaggcag cattgctgtt cgattcgtta ccaattgaca
      121 gctagctcag tcctaggtat atacatacat gcttgtttgt ttgtaaacta ctagagaagc
      181 gggcccaagt tcacttaaaa aggagatcaa caatgaaagc aattttcgta ctgaaacatc
      241 ttaatcatgc tggggagggt ttctaatgcc gaaactgggt atgcagagca ttcgtcgtcg
      301 tcagctgatt gatgcaaccc tggaagcaat taatgaagtt ggtatgcatg atgcaaccat
      361 tgcacagatt gcacgtcgtg ccggtgttag caccggtatt attagccatt atttccgcga
      421 taaaaacggt ctgctggaag caaccatgcg tgatattacc agccagctgc gtgatgcagt
      481 tctgaatcgt ctgcatgcac tgccgcaggg tagcgcagaa cagcgtctgc aggcaattgt
      541 tggtggtaat tttgatgaaa cccaggttag cagcgcagca atgaaagcat ggctggcatt
      601 ttgggcaagc agcatgcatc agccgatgct gtatcgtctg cagcaggtta gcagtcgtcg
      661 tctgctgagc aatctggtta gcgaatttcg tcgtgaactg cctcgtgaac aggcacaaga
      721 ggcaggttat ggtctggcag cactgattga tggtctgtgg ctgcgtgcag cactgagcgg
      781 taaaccgctg gataaaaccc gtgcaaatag cctgacccgt cattttatca cccagcatct
      841 gccgaccgat gcagcaaacg acgaaaacta cgctttagca gcttaatgaa ctaacgcatg
      901 agaaagcccc cggaagatca ccttccgggg gcttttttat tgcgcccttg agagtcgatg
      961 tttgtcttct aaggatccaa actcgagtaa ggatctccag gcatcaaata aaacgaaagg
     1021 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact
     1081 agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttatacctag ggtacgggtt
     1141 ttgctgcccg caaacgggct gttctggtgt tgctagtttg ttatcagaat cgcagatccg
     1201 gcttcagccg gtttgccggc tgaaagcgct atttcttcca gaattgccat gattttttcc
     1261 ccacgggagg cgtcactggc tcccgtgttg tcggcagctt tgattcgata agcagcatcg
     1321 cctgtttcag gctgtctatg tgtgactgtt gagctgtaac aagttgtctc aggtgttcaa
     1381 tttcatgttc tagttgcttt gttttactgg tttcacctgt tctattaggt gttacatgct
     1441 gttcatctgt tacattgtcg atctgttcat ggtgaacagc tttgaatgca ccaaaaactc
     1501 gtaaaagctc tgatgtatct atctttttta caccgttttc atctgtgcat atggacagtt
     1561 ttccctttga tatgtaacgg tgaacagttg ttctactttt gtttgttagt cttgatgctt
     1621 cactgataga tacaagagcc ataagaacct cagatccttc cgtatttagc cagtatgttc
     1681 tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg aaccattgag atcatactta
     1741 ctttgcatgt cactcaaaaa ttttgcctca aaactggtga gctgaatttt tgcagttaaa
     1801 gcatcgtgta gtgtttttct tagtccgtta tgtaggtagg aatctgatgt aatggttgtt
     1861 ggtattttgt caccattcat ttttatctgg ttgttctcaa gttcggttac gagatccatt
     1921 tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg ggcggcctcg cttatcaacc
     1981 accaatttca tattgctgta agtgtttaaa tctttactta ttggtttcaa aacccattgg
     2041 ttaagccttt taaactcatg gtagttattt tcaagcatta acatgaactt aaattcatca
     2101 aggctaatct ctatatttgc cttgtgagtt ttcttttgtg ttagttcttt taataaccac
     2161 tcataaatcc tcatagagta tttgttttca aaagacttaa catgttccag attatatttt
     2221 atgaattttt ttaactggaa aagataaggc aatatctctt cactaaaaac taattctaat
     2281 ttttcgcttg agaacttggc atagtttgtc cactggaaaa tctcaaagcc tttaaccaaa
     2341 ggattcctga tttccacagt tctcgtcatc agctctctgg ttgctttagc taatacacca
     2401 taagcatttt ccctactgat gttcatcatc tgagcgtatt ggttataagt gaacgatacc
     2461 gtccgttctt tccttgtagg gttttcaatc gtggggttga gtagtgccac acagcataaa
     2521 attagcttgg tttcatgctc cgttaagtca tagcgactaa tcgctagttc atttgctttg
     2581 aaaacaacta attcagacat acatctcaat tggtctaggt gattttaatc actataccaa
     2641 ttgagatggg ctagtcaatg ataattacta gtccttttcc cgggtgatct gggtatctgt
     2701 aaattctgct agacctttgc tggaaaactt gtaaattctg ctagaccctc tgtaaattcc
     2761 gctagacctt tgtgtgtttt ttttgtttat attcaagtgg ttataattta tagaataaag
     2821 aaagaataaa aaaagataaa aagaatagat cccagccctg tgtataactc actactttag
     2881 tcagttccgc agtattacaa aaggatgtcg caaacgctgt ttgctcctct acaaaacaga
     2941 ccttaaaacc ctaaaggctt aagtagcacc ctcgcaagct cgggcaaatc gctgaatatt
     3001 ccttttgtct ccgaccatca ggcacctgag tcgctgtctt tttcgtgaca ttcagttcgc
     3061 tgcgctcacg gctctggcag tgaatggggg taaatggcac tacaggcgcc ttttatggat
     3121 tcatgcaagg aaactaccca taatacaaga aaagcccgtc acgggcttct cagggcgttt
     3181 tatggcgggt ctgctatgtg gtgctatctg actttttgct gttcagcagt tcctgccctc
     3241 tgattttcca gtctgaccac ttcggattat cccgtgacag gtcattcaga ctggctaatg
     3301 cacccagtaa ggcagcggta tcatcaacag gcttacccgt cttactgtcc ctagtgcttg
     3361 gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga
     3421 gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctctcg aaccccagag
     3481 tcccgcttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3541 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3601 tggccccagt gctgcaatga taccgcgtga cccacgctca ccggctccag atttatcagc
     3661 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3721 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3781 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3841 ttcattcagc tccggttccc aacgatcaag gcgagttgca tgatccccca tgttgtgcaa
     3901 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3961 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4021 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4081 gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa
     4141 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4201 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4261 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4321 ggcgacacgg aaatgttgaa tactcatgcg aaacgatcct catcctgtct cttgatcaga
     4381 tcatgatccc ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca
     4441 gggcttccca accttaccag agggcgcccc agctggcaat tcc
//
