LOCUS       sequence-457233-        6823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR array plasmid for NOTCH2 (Spacers 25-48).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6823)
  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 6823)
  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: pCX138
FEATURES             Location/Qualifiers
     source          1..6823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3845..3864)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3910..3965
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3964..3983)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4326..4342)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4326..4342)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4339..4361)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4350..4366
                     /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(4374..4380,4381..4398,4399..4404))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4419..4440
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4499..4694
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4545..4680
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4700..4834
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4737..4756)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4791..4810
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4902..4919)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5073..5661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5153..5172)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5832..6623,6624..6692))
                     /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     6455..6474
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6693..6797)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gccagggggg gggggggggg gggagggggg
      541 ggggggggsg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcck cgggccgggg rgggctcggg ggrggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcgccaccgt
     1741 ggactcagcg gcagagacgc caccccgaga aggcaaatcc cactttcaga tcctgctgga
     1801 cgtggtccag ttcctccctg aagacatcat cattcagacc ttcgaaggct ggctgctgat
     1861 aaaagcacaa cacggaacca gaatggatga gcacggtttt atctcaagaa gcttcacccg
     1921 acagtacaaa ctaccagatg gtgtggaaat caaagatttg tctgcagtcc tctgtcatga
     1981 tggaattttg gtggtggaag taaaggatcc agttgggact aaggagctca tcgaatcctt
     2041 aagctacgaa cgatataaac ctaattacct cgagagggga cggaaacaaa gagtcgggaa
     2101 ttcacctgtt aatatctaca aaagatataa acctaattac ctcgagaggg gacggaaaca
     2161 taggactgaa aaaaccattt gtaaactatt atgccgatat aaacctaatt acctcgagag
     2221 gggacggaaa caaatcaggt aagtgggaag cactgatgca tacttgcgat ataaacctaa
     2281 ttacctcgag aggggacgga aacaaatggg tccaagggca gagagtcacc atttagaatg
     2341 atataaacct aattacctcg agaggggacg gaaacaaagc agagaatgaa caaacttaat
     2401 ctgcttatct cgatataaac ctaattacct cgagagggga cggaaacagg gcagaaaggg
     2461 gcaggtacgc tgtggtccac agagatataa acctaattac ctcgagaggg gacggaaaca
     2521 aaagggagga tataaaatat ccagtggctg gatcagatat aaacctaatt acctcgagag
     2581 gggacggaaa catggcataa aagatgtgtc tcataaaaac tgagtctgat ataaacctaa
     2641 ttacctcgag aggggacgga aacaaaggca gtgttgtcct cctcatcact gaaacctgtg
     2701 atataaacct aattacctcg agaggggacg gaaacaaacc caaaggatcc aaattaaagt
     2761 agtccaagaa agatataaac ctaattacct cgagagggga cggaaactta ccttcccttt
     2821 tattagatac cacacaattc aaggatataa acctaattac ctcgagaggg gacggaaaca
     2881 gaagaggagt caaagaaaga aagcatttat aacaagatat aaacctaatt acctcgagag
     2941 gggacggaaa cacctcacat aagaaaatga tgcttaaaac aaaacaggat ataaacctaa
     3001 ttacctcgag aggggacgga aacaaaacat tacacctttg gttctttatt atgcaaaaag
     3061 atataaacct aattacctcg agaggggacg gaaactttga gcatcacagc caattgctta
     3121 tactaaaata tgatataaac ctaattacct cgagagggga cggaaacaaa tgtgtcctcc
     3181 caaatagtca tcatacattc aatgatataa acctaattac ctcgagaggg gacggaaaca
     3241 aagtgtgcat tttccttact acgtttagtc aggaagatat aaacctaatt acctcgagag
     3301 gggacggaaa catatcactt ttaagagaaa tgtacacaag gaagtaagat ataaacctaa
     3361 ttacctcgag aggggacgga aacttttgtg ataaaaatgc tttcatataa atttcatctg
     3421 atataaacct aattacctcg agaggggacg gaaactttgt gggattcaga aagaaaagaa
     3481 aatttgctct cgatataaac ctaattacct cgagagggga cggaaactgc caggccatgg
     3541 atgcagtatt ggaaaataaa aaagatataa acctaattac ctcgagaggg gacggaaact
     3601 ttccccagga tcatttattt atgatctaat taaaggatat aaacctaatt acctcgagag
     3661 gggacggaaa cttcccaatt cttgctatag caactacttc gcatttcgat ataaacctaa
     3721 ttacctcgag aggggacgga aactttgtca ccagctatgg ctagtgcaca gaagagtgcg
     3781 atataaacct aattacctcg agaggggacg gaaacaggtt aacactgccc ttgaagctag
     3841 cagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga
     3901 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     3961 tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg
     4021 tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa
     4081 gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct
     4141 tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc
     4201 taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag
     4261 tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag cttggcgtaa
     4321 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4381 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4441 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc
     4501 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4561 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4621 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4681 taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa
     4741 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4801 caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc
     4861 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4921 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4981 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5041 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5101 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5161 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5221 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5281 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5341 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5401 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5461 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5521 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5581 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5641 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5701 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5761 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5821 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5881 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5941 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6001 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6061 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6121 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6181 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6241 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6301 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6361 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6421 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6481 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6541 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6601 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6661 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6721 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6781 caaatagggg ttccgcgcac atttccccga aaagtgccac ctg
//
