LOCUS       sequence-311610-        2683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dCas9 sgRNA oscillator v1 (dCRv1) (p15A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2683)
  AUTHORS   Kuo J, Yuan R, Sanchez C, Paulsson J, Silver PA
  TITLE     Toward a translationally independent RNA-based synthetic oscillator 
            using deactivated CRISPR-Cas.
  JOURNAL   Nucleic Acids Res. 2020 Aug 20;48(14):8165-8177. doi: 
            10.1093/nar/gkaa557.
  PUBMED    32609820
REFERENCE   2  (bases 1 to 2683)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2020-08-20- 20"; volume: "48"; issue: 
            "14"; pages: "8165-8177"
COMMENT     Sequence Label: pJK398
FEATURES             Location/Qualifiers
     source          1..2683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     64..83
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(302..406)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     474..492
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(496..539)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_RNA        622..697
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        981..1015
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     misc_RNA        1036..1111
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1197..1214)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1331..1876)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1411..1430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2124..2683,1..232,233..301))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
       61 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
      121 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
      181 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
      241 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
      301 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
      361 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
      421 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
      481 gtatcacgag gcagaatttc agataaaaaa aatccttagc tttcgctaag gatgatttct
      541 ggaattctaa agatctcaga aatcatttta caccaacggg tcacacgggt attatgctag
      601 cataatacca gctagttgtg ggttttagag ctagaaatag caagttaaaa taaggctagt
      661 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttctcggt accaaattcc
      721 agaaaagagg cctcccgaaa ggggggcctt ttttcgtttt ggtccgctta agctttttac
      781 acctccacaa ctagctggta ttatgctagc ataataccta ggactgagct gttttagagc
      841 tagaaatagc aagttaaaat aaggctagtc cgattgcaac ttgaaaaagt ggcaccgagt
      901 cggtgctttt tttctcggta ccaaattcca gaaaagagac gctttcgagc gtcttttttc
      961 gttttggtcc gataggatcc tttacaccta gctcagtcct aggtattatg ctagcataat
     1021 acccgtgtga cccgtgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
     1081 tcaacttgaa aaagtggcac cgagtcggtg ctttttttct cggtaccaaa gacgaacaat
     1141 aagacgctga aaagcgtctt ttttcgtttt ggtcctttat ctagactgca ggcttcctcg
     1201 ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg
     1261 ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg
     1321 caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa
     1381 atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct
     1441 ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc
     1501 cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg
     1561 gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt
     1621 cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga
     1681 tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt
     1741 tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac
     1801 cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag
     1861 accaaaacga tctcaagaag atcatcttat taatcagata aaatatttct agatttcagt
     1921 gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata agttgttact
     1981 agtgcttggc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2041 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2101 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2161 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2221 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2281 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2341 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2401 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     2461 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2521 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2581 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2641 atgccatccg taagatgctt ttctgtgact ggtgagtact caa
//
