LOCUS       sequence-126636-        4555 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 4555)
  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 4555)
  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: pQacR-BCD7-tetR-ssrA P68U18C45T13 v40-5 (569)
FEATURES             Location/Qualifiers
     source          1..4555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             265..885
                     /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(338..357)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      1077..1148
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1164..1191
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1722..2672)
                     /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(3434..3528)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(3559..4350,4351..4419))
                     /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     4182..4201
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gacgtcggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
       61 ttccagtcgg gaaacctgaa gacaaaaccg cattgctgtt cgattcgtta ccaattgaca
      121 gctagctcag tcctacttta gtatagagac tgagcggtcg gtctatatac tagagaagcg
      181 ggcccaagtt cacttaaaaa ggagatcaac aatgaaagca attttcgtac tgaaacatct
      241 taatcatgct ggggagggtt tctaatgtct agattagata aaagtaaagt gattaacagc
      301 gcattagagc tgcttaatga ggtcggaatc gaaggtttaa caacccgtaa actcgcccag
      361 aagctaggtg tagagcagcc tacattgtat tggcatgtaa aaaataagcg ggctttgctc
      421 gacgccttag ccattgagat gttagatagg caccatactc acttttgccc tttagaaggg
      481 gaaagctggc aagatttttt acgtaataac gctaaaagtt ttagatgtgc tttactaagt
      541 catcgcgatg gagcaaaagt acatttaggt acacggccta cagaaaaaca gtatgaaact
      601 ctcgaaaatc aattagcctt tttatgccaa caaggttttt cactagagaa tgcattatat
      661 gcactcagcg ctgtggggca ttttacttta ggttgcgtat tggaagatca agagcatcaa
      721 gtcgctaaag aagaaaggga aacacctact actgatagta tgccgccatt attacgacaa
      781 gctatcgaat tatttgatca ccaaggtgca gagccagcct tcttattcgg ccttgaattg
      841 atcatatgcg gattagaaaa acaacttaaa tgtgaaagtg ggtctgcagc aaacgacgaa
      901 aactacgctt tagcagctta atgaactttc agccaaaaaa cttaagaccg ccggtcttgt
      961 ccactacctt gcagtaatgc ggtggacagg atcggcggtt ttcttttctc ttctcaacct
     1021 tgagagtcgg cttttgtctt ctaaggatcc aaactcgagt aaggatctcc aggcatcaaa
     1081 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     1141 acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatacct
     1201 agggtacggg ttttgctgcc cgcaaacggg ctgttctggt gttgctagtt tgttatcaga
     1261 atcgcagatc cggcttcagc cggtttgccg gctgaaagcg ctatttcttc cagaattgcc
     1321 atgatttttt ccccacggga ggcgtcactg gctcccgtgt tgtcggcagc tttgattcga
     1381 taagcagcat cgcctgtttc aggctgtcta tgtgtgactg ttgagctgta acaagttgtc
     1441 tcaggtgttc aatttcatgt tctagttgct ttgttttact ggtttcacct gttctattag
     1501 gtgttacatg ctgttcatct gttacattgt cgatctgttc atggtgaaca gctttgaatg
     1561 caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt tacaccgttt tcatctgtgc
     1621 atatggacag ttttcccttt gatatgtaac ggtgaacagt tgttctactt ttgtttgtta
     1681 gtcttgatgc ttcactgata gatacaagag ccataagaac ctcagatcct tccgtattta
     1741 gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg
     1801 agatcatact tactttgcat gtcactcaaa aattttgcct caaaactggt gagctgaatt
     1861 tttgcagtta aagcatcgtg tagtgttttt cttagtccgt tatgtaggta ggaatctgat
     1921 gtaatggttg ttggtatttt gtcaccattc atttttatct ggttgttctc aagttcggtt
     1981 acgagatcca tttgtctatc tagttcaact tggaaaatca acgtatcagt cgggcggcct
     2041 cgcttatcaa ccaccaattt catattgctg taagtgttta aatctttact tattggtttc
     2101 aaaacccatt ggttaagcct tttaaactca tggtagttat tttcaagcat taacatgaac
     2161 ttaaattcat caaggctaat ctctatattt gccttgtgag ttttcttttg tgttagttct
     2221 tttaataacc actcataaat cctcatagag tatttgtttt caaaagactt aacatgttcc
     2281 agattatatt ttatgaattt ttttaactgg aaaagataag gcaatatctc ttcactaaaa
     2341 actaattcta atttttcgct tgagaacttg gcatagtttg tccactggaa aatctcaaag
     2401 cctttaacca aaggattcct gatttccaca gttctcgtca tcagctctct ggttgcttta
     2461 gctaatacac cataagcatt ttccctactg atgttcatca tctgagcgta ttggttataa
     2521 gtgaacgata ccgtccgttc tttccttgta gggttttcaa tcgtggggtt gagtagtgcc
     2581 acacagcata aaattagctt ggtttcatgc tccgttaagt catagcgact aatcgctagt
     2641 tcatttgctt tgaaaacaac taattcagac atacatctca attggtctag gtgattttaa
     2701 tcactatacc aattgagatg ggctagtcaa tgataattac tagtcctttt cccgggtgat
     2761 ctgggtatct gtaaattctg ctagaccttt gctggaaaac ttgtaaattc tgctagaccc
     2821 tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt atattcaagt ggttataatt
     2881 tatagaataa agaaagaata aaaaaagata aaaagaatag atcccagccc tgtgtataac
     2941 tcactacttt agtcagttcc gcagtattac aaaaggatgt cgcaaacgct gtttgctcct
     3001 ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag ctcgggcaaa
     3061 tcgctgaata ttccttttgt ctccgaccat caggcacctg agtcgctgtc tttttcgtga
     3121 cattcagttc gctgcgctca cggctctggc agtgaatggg ggtaaatggc actacaggcg
     3181 ccttttatgg attcatgcaa ggaaactacc cataatacaa gaaaagcccg tcacgggctt
     3241 ctcagggcgt tttatggcgg gtctgctatg tggtgctatc tgactttttg ctgttcagca
     3301 gttcctgccc tctgattttc cagtctgacc acttcggatt atcccgtgac aggtcattca
     3361 gactggctaa tgcacccagt aaggcagcgg tatcatcaac aggcttaccc gtcttactgt
     3421 ccctagtgct tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca
     3481 aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctct
     3541 cgaaccccag agtcccgctt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3601 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3661 gggcttacca tctggcccca gtgctgcaat gataccgcgt gacccacgct caccggctcc
     3721 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3781 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3841 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3901 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagttg catgatcccc
     3961 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4021 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4081 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4141 tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag
     4201 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4261 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4321 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4381 aaagggaata agggcgacac ggaaatgttg aatactcatg cgaaacgatc ctcatcctgt
     4441 ctcttgatca gatcatgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca
     4501 gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca attcc
//
