LOCUS       sequence-457234-        6822 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR array plasmid for NOTCH2 (Spacers 1-24).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6822)
  AUTHORS   Xia C, Colognori D, Jiang XS, Xu K, Doudna JA
  TITLE     Single-molecule live-cell RNA imaging with CRISPR-Csm.
  JOURNAL   Nat Biotechnol. 2025 Feb 18. doi: 10.1038/s41587-024-02540-5.
  PUBMED    39966655
REFERENCE   2  (bases 1 to 6822)
  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: "Nat 
            Biotechnol. 2025 Feb 18. doi: 10.1038/s41587-024-02540-5."
COMMENT     Sequence Label: pCX137
FEATURES             Location/Qualifiers
     source          1..6822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..395
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        397..672
                     /label=chicken beta-actin promoter
     intron          673..1689
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1613..1635
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1697..1716
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3855..3874)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3920..3975
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3974..3993)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4336..4352)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4336..4352)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4349..4371)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4360..4376
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4384..4390,4391..4408,4409..4414))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4429..4450
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4509..4704
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4555..4690
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4710..4844
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4747..4766)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4801..4820
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4912..4929)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5083..5671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5163..5182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5842..6633,6634..6702))
                     /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"
     primer_bind     6465..6484
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6703..6807)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agtgccacct gggtcgacat tgattattga ctagttatta atagtaatca attacggggt
       61 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      121 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      181 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      241 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      301 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      361 agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct
      421 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat
      481 tattttgtgc agcgatgggg gcgggggggg ggggggggcg cgccaggggg gggggggggg
      541 ggggaggggg gggggggggs gaggsggaga ggtgcggcgg cagccaatca gagcggcgcg
      601 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
      661 gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc
      721 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
      781 ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg
      841 tggctgcgtg aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg
      901 gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc
      961 ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg
     1021 agcgcggccg ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc
     1081 gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa
     1141 ccccccctgc acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc
     1201 cgtacggggc gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg
     1261 ccgggcgggg cggggccgcc kcgggccggg grgggctcgg gggrggggcg cggcggcccc
     1321 cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg
     1381 tgcgagaggg cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg
     1441 cgccgccgca ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa
     1501 atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct
     1561 cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc
     1621 ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt
     1681 ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa
     1741 ttcgccaccg tggactcagc ggcagagacg ccaccccgag aaggcaaatc ccactttcag
     1801 atcctgctgg acgtggtcca gttcctccct gaagacatca tcattcagac cttcgaaggc
     1861 tggctgctga taaaagcaca acacggaacc agaatggatg agcacggttt tatctcaaga
     1921 agcttcaccc gacagtacaa actaccagat ggtgtggaaa tcaaagattt gtctgcagtc
     1981 ctctgtcatg atggaatttt ggtggtggaa gtaaaggatc cagttgggac taaggagctc
     2041 atcgaatcct taagctacga acgatataaa cctaattacc tcgagagggg acggaaactt
     2101 tgttcctcag cagcatttac aaaagtcagt tatggatata aacctaatta cctcgagagg
     2161 ggacggaaac tacccagtga aactgacgaa ttgcttctct tgcattgata taaacctaat
     2221 tacctcgaga ggggacggaa actacctcta gaagctggct ccagagattt cttcatttga
     2281 tataaaccta attacctcga gaggggacgg aaacttgtat tcatcttgca tttccacaaa
     2341 cttgtcttat gatataaacc taattacctc gagaggggac ggaaacttca ataagcatcc
     2401 atcttattct ccaaatagaa gagatataaa cctaattacc tcgagagggg acggaaactt
     2461 ctcatggata tggcagaagc ctgcagggct taaagatata aacctaatta cctcgagagg
     2521 ggacggaaac tttcatgggc cagaatgatc aaaagcacaa aggtctgata taaacctaat
     2581 tacctcgaga ggggacggaa actttatatg aagacctgca cacagacaag aaaaattaga
     2641 tataaaccta attacctcga gaggggacgg aaactttgtg ctgagaacca taccaaagta
     2701 aaccttgtga gatataaacc taattacctc gagaggggac ggaaacttct ctctccagga
     2761 atgatatatg caggagaaca cagatataaa cctaattacc tcgagagggg acggaaactt
     2821 ccaaaccttt ttctggtttt aaaaaagtgg ggaggatata aacctaatta cctcgagagg
     2881 ggacggaaac tttacactga ccaggcccat acacagaaaa acaacagata taaacctaat
     2941 tacctcgaga ggggacggaa acaaacaggc ctagaatgct cagatctggg acaaatgtga
     3001 tataaaccta attacctcga gaggggacgg aaacttggca ggaagtaggg ggacctgtgg
     3061 gtgggtaact gatataaacc taattacctc gagaggggac ggaaacttct gtggatatga
     3121 aaggaaaggc tcaacagtta aggatataaa cctaattacc tcgagagggg acggaaactt
     3181 ggtgagcagg cctttaagca gcaagataat cagtgatata aacctaatta cctcgagagg
     3241 ggacggaaac tgtcactggg tcccttgggc taaggaaagt tcagtcgata taaacctaat
     3301 tacctcgaga ggggacggaa actataaagt ccatgtcttc agtgagaaca tactgggtga
     3361 tataaaccta attacctcga gaggggacgg aaacttctta tagggctgtt ttggatcata
     3421 gaaaacaagt gatataaacc taattacctc gagaggggac ggaaactagg aaaggaaaat
     3481 gttctgttga gtttctacgt tagatataaa cctaattacc tcgagagggg acggaaactt
     3541 cagaggaaaa gaaatggtag cagaaaatag caaagatata aacctaatta cctcgagagg
     3601 ggacggaaac ttcaggtgag gggcaatcag tctgaacaat gagcaagata taaacctaat
     3661 tacctcgaga ggggacggaa acttaactaa aatgcttctg cccttaaaag aaagtcaaga
     3721 tataaaccta attacctcga gaggggacgg aaacttgcag tgtagccatg gaccaattat
     3781 taactggccc gatataaacc taattacctc gagaggggac ggaaacaccg gtgcaattac
     3841 gcttagctag cagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     3901 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     3961 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     4021 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     4081 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     4141 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     4201 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     4261 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag
     4321 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4381 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4441 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4501 gcggatccgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4561 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4621 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4681 tttggaggcc taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca
     4741 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4801 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat
     4861 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4921 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4981 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5041 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5101 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5161 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5221 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5281 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5341 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5401 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5461 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5521 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5581 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5641 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5701 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5761 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5821 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5881 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5941 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6001 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6061 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6121 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6181 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6241 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6301 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6361 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6421 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6481 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6541 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6601 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6661 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6721 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6781 gaaaaataaa caaatagggg ttccgcgcac atttccccga aa
//
