LOCUS       sequence-126632-        4519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid for building 3n2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4519)
  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 4519)
  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-tetR-ssrA P65U18C45T14 v40-2 (491)
FEATURES             Location/Qualifiers
     source          1..4519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(137..205,206..997)
                     /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     complement(355..374)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      1028..1122
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1884..2834
                     /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(3365..3392)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(3408..3479)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             3703..4323
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(3776..3795)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 ggaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat
       61 ggctttcttg ccgccaagga tctgatggcg caggggatca tgatctgatc aagagacagg
      121 atgaggatcg tttcgcatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
      181 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
      241 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
      301 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      361 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      421 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
      481 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
      541 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
      601 tcatgcaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
      661 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
      721 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
      781 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
      841 cggtgagcgt gggtcacgcg gtatcattgc agcactgggg ccagatggta agccctcccg
      901 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
      961 cgctgagata ggtgcctcac tgattaagca ttggtaagcg ggactctggg gttcgagagc
     1021 tcgcttggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
     1081 agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcac tagggacagt
     1141 aagacgggta agcctgttga tgataccgct gccttactgg gtgcattagc cagtctgaat
     1201 gacctgtcac gggataatcc gaagtggtca gactggaaaa tcagagggca ggaactgctg
     1261 aacagcaaaa agtcagatag caccacatag cagacccgcc ataaaacgcc ctgagaagcc
     1321 cgtgacgggc ttttcttgta ttatgggtag tttccttgca tgaatccata aaaggcgcct
     1381 gtagtgccat ttacccccat tcactgccag agccgtgagc gcagcgaact gaatgtcacg
     1441 aaaaagacag cgactcaggt gcctgatggt cggagacaaa aggaatattc agcgatttgc
     1501 ccgagcttgc gagggtgcta cttaagcctt tagggtttta aggtctgttt tgtagaggag
     1561 caaacagcgt ttgcgacatc cttttgtaat actgcggaac tgactaaagt agtgagttat
     1621 acacagggct gggatctatt ctttttatct ttttttattc tttctttatt ctataaatta
     1681 taaccacttg aatataaaca aaaaaaacac acaaaggtct agcggaattt acagagggtc
     1741 tagcagaatt tacaagtttt ccagcaaagg tctagcagaa tttacagata cccagatcac
     1801 ccgggaaaag gactagtaat tatcattgac tagcccatct caattggtat agtgattaaa
     1861 atcacctaga ccaattgaga tgtatgtctg aattagttgt tttcaaagca aatgaactag
     1921 cgattagtcg ctatgactta acggagcatg aaaccaagct aattttatgc tgtgtggcac
     1981 tactcaaccc cacgattgaa aaccctacaa ggaaagaacg gacggtatcg ttcacttata
     2041 accaatacgc tcagatgatg aacatcagta gggaaaatgc ttatggtgta ttagctaaag
     2101 caaccagaga gctgatgacg agaactgtgg aaatcaggaa tcctttggtt aaaggctttg
     2161 agattttcca gtggacaaac tatgccaagt tctcaagcga aaaattagaa ttagttttta
     2221 gtgaagagat attgccttat cttttccagt taaaaaaatt cataaaatat aatctggaac
     2281 atgttaagtc ttttgaaaac aaatactcta tgaggattta tgagtggtta ttaaaagaac
     2341 taacacaaaa gaaaactcac aaggcaaata tagagattag ccttgatgaa tttaagttca
     2401 tgttaatgct tgaaaataac taccatgagt ttaaaaggct taaccaatgg gttttgaaac
     2461 caataagtaa agatttaaac acttacagca atatgaaatt ggtggttgat aagcgaggcc
     2521 gcccgactga tacgttgatt ttccaagttg aactagatag acaaatggat ctcgtaaccg
     2581 aacttgagaa caaccagata aaaatgaatg gtgacaaaat accaacaacc attacatcag
     2641 attcctacct acataacgga ctaagaaaaa cactacacga tgctttaact gcaaaaattc
     2701 agctcaccag ttttgaggca aaatttttga gtgacatgca aagtaagtat gatctcaatg
     2761 gttcgttctc atggctcacg caaaaacaac gaaccacact agagaacata ctggctaaat
     2821 acggaaggat ctgaggttct tatggctctt gtatctatca gtgaagcatc aagactaaca
     2881 aacaaaagta gaacaactgt tcaccgttac atatcaaagg gaaaactgtc catatgcaca
     2941 gatgaaaacg gtgtaaaaaa gatagataca tcagagcttt tacgagtttt tggtgcattc
     3001 aaagctgttc accatgaaca gatcgacaat gtaacagatg aacagcatgt aacacctaat
     3061 agaacaggtg aaaccagtaa aacaaagcaa ctagaacatg aaattgaaca cctgagacaa
     3121 cttgttacag ctcaacagtc acacatagac agcctgaaac aggcgatgct gcttatcgaa
     3181 tcaaagctgc cgacaacacg ggagccagtg acgcctcccg tggggaaaaa atcatggcaa
     3241 ttctggaaga aatagcgctt tcagccggca aaccggctga agccggatct gcgattctga
     3301 taacaaacta gcaacaccag aacagcccgt ttgcgggcag caaaacccgt accctaggta
     3361 taaacgcaga aaggcccacc cgaaggtgag ccagtgtgac tctagtagag agcgttcacc
     3421 gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga
     3481 tgcctggaga tccttactcg agtttggatc cttagaagac aaacatgcat tgctgttcga
     3541 ttcgttacca attgacagct agctcagtcc taggtatata catacatgct tgtttgtttg
     3601 taaactacta gagaagcggg cccaagttca cttaaaaagg agatcaacaa tgaaagcaat
     3661 tttcgtactg aaacatctta atcatgctgg ggagggtttc taatgtctag attagataaa
     3721 agtaaagtga ttaacagcgc attagagctg cttaatgagg tcggaatcga aggtttaaca
     3781 acccgtaaac tcgcccagaa gctaggtgta gagcagccta cattgtattg gcatgtaaaa
     3841 aataagcggg ctttgctcga cgccttagcc attgagatgt tagataggca ccatactcac
     3901 ttttgccctt tagaagggga aagctggcaa gattttttac gtaataacgc taaaagtttt
     3961 agatgtgctt tactaagtca tcgcgatgga gcaaaagtac atttaggtac acggcctaca
     4021 gaaaaacagt atgaaactct cgaaaatcaa ttagcctttt tatgccaaca aggtttttca
     4081 ctagagaatg cattatatgc actcagcgct gtggggcatt ttactttagg ttgcgtattg
     4141 gaagatcaag agcatcaagt cgctaaagaa gaaagggaaa cacctactac tgatagtatg
     4201 ccgccattat tacgacaagc tatcgaatta tttgatcacc aaggtgcaga gccagccttc
     4261 ttattcggcc ttgaattgat catatgcgga ttagaaaaac aacttaaatg tgaaagtggg
     4321 tctgcagcaa acgacgaaaa ctacgcttta gcagcttaat gaactaacgc atgagaaagc
     4381 ccccggaaga tcaccttccg ggggcttttt tattgcgccc ttgagagtcg tgccttgtct
     4441 tcaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca
     4501 ctcattaggc accgacgtc
//
