LOCUS       sequence-126626-        4475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid for building 3n1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4475)
  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 4475)
  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-phlF-ssrA P69U18C51T15 v40-3 (570)
FEATURES             Location/Qualifiers
     source          1..4475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      997..1068
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1084..1111
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1642..2592)
                     /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(3354..3448)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(3479..4270,4271..4339))
                     /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     4102..4121
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gacgtcggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
       61 ttccagtcgg gaaacctgaa gacaaatgtg cattgctgtt cgattcgtta ccaattgaca
      121 attgattgga cgttcaatat aatgctagct actagagaag cgggcccaag ttcacttaaa
      181 aaggagatca acaatgaaag caattttcgt actgaaacat cttaatcatg ctggggaggg
      241 tttctaatgg cacgtacccc gagccgtagc agcattggta gcctgcgtag tccgcatacc
      301 cataaagcaa ttctgaccag caccattgaa atcctgaaag aatgtggtta tagcggtctg
      361 agcattgaaa gcgttgcacg tcgtgccggt gcaagcaaac cgaccattta tcgttggtgg
      421 accaataaag cagcactgat tgccgaagtg tatgaaaatg aaagcgaaca ggtgcgtaaa
      481 tttccggatc tgggtagctt taaagccgat ctggattttc tgctgcgtaa tctgtggaaa
      541 gtttggcgtg aaaccatttg tggtgaagca tttcgttgtg ttattgcaga agcacagctg
      601 gaccctgcaa ccctgaccca gctgaaagat cagtttatgg aacgtcgtcg tgagatgccg
      661 aaaaaactgg ttgaaaatgc cattagcaat ggtgaactgc cgaaagatac caatcgtgaa
      721 ctgctgctgg atatgatttt tggtttttgt tggtatcgcc tgctgaccga acagctgacc
      781 gttgaacagg atattgaaga atttaccttc ctgctgatta atggtgtttg tccgggtaca
      841 cagcgtgcag caaacgacga aaactacgct ttagcagctt aatgaacttc cggcaattaa
      901 aaaagcggct aaccacgccg ctttttttac gtctgcacct tgagagtcgc tcgttgtctt
      961 ctaaggatcc aaactcgagt aaggatctcc aggcatcaaa taaaacgaaa ggctcagtcg
     1021 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac
     1081 actggctcac cttcgggtgg gcctttctgc gtttatacct agggtacggg ttttgctgcc
     1141 cgcaaacggg ctgttctggt gttgctagtt tgttatcaga atcgcagatc cggcttcagc
     1201 cggtttgccg gctgaaagcg ctatttcttc cagaattgcc atgatttttt ccccacggga
     1261 ggcgtcactg gctcccgtgt tgtcggcagc tttgattcga taagcagcat cgcctgtttc
     1321 aggctgtcta tgtgtgactg ttgagctgta acaagttgtc tcaggtgttc aatttcatgt
     1381 tctagttgct ttgttttact ggtttcacct gttctattag gtgttacatg ctgttcatct
     1441 gttacattgt cgatctgttc atggtgaaca gctttgaatg caccaaaaac tcgtaaaagc
     1501 tctgatgtat ctatcttttt tacaccgttt tcatctgtgc atatggacag ttttcccttt
     1561 gatatgtaac ggtgaacagt tgttctactt ttgtttgtta gtcttgatgc ttcactgata
     1621 gatacaagag ccataagaac ctcagatcct tccgtattta gccagtatgt tctctagtgt
     1681 ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg agatcatact tactttgcat
     1741 gtcactcaaa aattttgcct caaaactggt gagctgaatt tttgcagtta aagcatcgtg
     1801 tagtgttttt cttagtccgt tatgtaggta ggaatctgat gtaatggttg ttggtatttt
     1861 gtcaccattc atttttatct ggttgttctc aagttcggtt acgagatcca tttgtctatc
     1921 tagttcaact tggaaaatca acgtatcagt cgggcggcct cgcttatcaa ccaccaattt
     1981 catattgctg taagtgttta aatctttact tattggtttc aaaacccatt ggttaagcct
     2041 tttaaactca tggtagttat tttcaagcat taacatgaac ttaaattcat caaggctaat
     2101 ctctatattt gccttgtgag ttttcttttg tgttagttct tttaataacc actcataaat
     2161 cctcatagag tatttgtttt caaaagactt aacatgttcc agattatatt ttatgaattt
     2221 ttttaactgg aaaagataag gcaatatctc ttcactaaaa actaattcta atttttcgct
     2281 tgagaacttg gcatagtttg tccactggaa aatctcaaag cctttaacca aaggattcct
     2341 gatttccaca gttctcgtca tcagctctct ggttgcttta gctaatacac cataagcatt
     2401 ttccctactg atgttcatca tctgagcgta ttggttataa gtgaacgata ccgtccgttc
     2461 tttccttgta gggttttcaa tcgtggggtt gagtagtgcc acacagcata aaattagctt
     2521 ggtttcatgc tccgttaagt catagcgact aatcgctagt tcatttgctt tgaaaacaac
     2581 taattcagac atacatctca attggtctag gtgattttaa tcactatacc aattgagatg
     2641 ggctagtcaa tgataattac tagtcctttt cccgggtgat ctgggtatct gtaaattctg
     2701 ctagaccttt gctggaaaac ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc
     2761 tttgtgtgtt ttttttgttt atattcaagt ggttataatt tatagaataa agaaagaata
     2821 aaaaaagata aaaagaatag atcccagccc tgtgtataac tcactacttt agtcagttcc
     2881 gcagtattac aaaaggatgt cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa
     2941 ccctaaaggc ttaagtagca ccctcgcaag ctcgggcaaa tcgctgaata ttccttttgt
     3001 ctccgaccat caggcacctg agtcgctgtc tttttcgtga cattcagttc gctgcgctca
     3061 cggctctggc agtgaatggg ggtaaatggc actacaggcg ccttttatgg attcatgcaa
     3121 ggaaactacc cataatacaa gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg
     3181 gtctgctatg tggtgctatc tgactttttg ctgttcagca gttcctgccc tctgattttc
     3241 cagtctgacc acttcggatt atcccgtgac aggtcattca gactggctaa tgcacccagt
     3301 aaggcagcgg tatcatcaac aggcttaccc gtcttactgt ccctagtgct tggattctca
     3361 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag
     3421 gtcattactg gatctatcaa caggagtcca agcgagctct cgaaccccag agtcccgctt
     3481 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3541 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3601 gtgctgcaat gataccgcgt gacccacgct caccggctcc agatttatca gcaataaacc
     3661 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3721 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3781 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3841 gctccggttc ccaacgatca aggcgagttg catgatcccc catgttgtgc aaaaaagcgg
     3901 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3961 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4021 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4081 cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4141 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4201 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4261 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4321 ggaaatgttg aatactcatg cgaaacgatc ctcatcctgt ctcttgatca gatcatgatc
     4381 ccctgcgcca tcagatcctt ggcggcaaga aagccatcca gtttactttg cagggcttcc
     4441 caaccttacc agagggcgcc ccagctggca attcc
//
