LOCUS       sequence-457230-        6727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR array plasmid for MAP1B (Spacers 25-48).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6727)
  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 6727)
  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: pCX460
FEATURES             Location/Qualifiers
     source          1..6727
                     /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(3749..3768)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3814..3869
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3868..3887)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4230..4246)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4230..4246)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4243..4265)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4254..4270
                     /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(4278..4284,4285..4302,4303..4308))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4323..4344
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4403..4598
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4449..4584
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4604..4738
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4641..4660)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4695..4714
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4806..4823)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4977..5565)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5057..5076)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5736..6527,6528..6596))
                     /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     6359..6378
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6597..6701)
                     /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 aggsggagag 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 cctacgttga
     2041 tgtatctctc agatataaac ctaattacct cgagagggga cggaaactct cccccactct
     2101 caagtagcac ttttatgttg atataaacct aattacctcg agaggggacg gaaacagaag
     2161 tatgtaggat accactgggt gatgtacgat ataaacctaa ttacctcgag aggggacgga
     2221 aacgaaccgt aggagttgag aattcatttg ttttcgatat aaacctaatt acctcgagag
     2281 gggacggaaa cgaaaatccc aaagcaaatt tgtgctaggg ttcgatataa acctaattac
     2341 ctcgagaggg gacggaaact aatcatgatc tggcttccca ggttcacagt tgatataaac
     2401 ctaattacct cgagagggga cggaaacacg gagaagtttg cctcatctga acactgggtg
     2461 atataaacct aattacctcg agaggggacg gaaactcatt gttctctact ccgtagatct
     2521 tagagtcgat ataaacctaa ttacctcgag aggggacgga aacgacttat cagacaaaaa
     2581 tcaactaaaa tgttagatat aaacctaatt acctcgagag gggacggaaa ctaggtcctc
     2641 agtgaaagga ccctgaagaa gcagatataa acctaattac ctcgagaggg gacggaaact
     2701 ttccaaggac ttgaaagaaa tggggtaata agatataaac ctaattacct cgagagggga
     2761 cggaaacata ttaattttct ttaaaaaatt aaacatttgg atataaacct aattacctcg
     2821 agaggggacg gaaacactca atgaaattca ttggcgtcac aatgactgat ataaacctaa
     2881 ttacctcgag aggggacgga aacccattaa ataattatct tgagtatctt aggttgatat
     2941 aaacctaatt acctcgagag gggacggaaa cacatttaat tgtgaaattg gaaaataact
     3001 tgcgatataa acctaattac ctcgagaggg gacggaaact tggcaagaag gctaaaaatc
     3061 tggttttctt cgatataaac ctaattacct cgagagggga cggaaacaac tgaagactgg
     3121 ctataattct cctacaatgg atataaacct aattacctcg agaggggacg gaaactttac
     3181 atatttttcc cattaaatcc catagcagat ataaacctaa ttacctcgag aggggacgga
     3241 aactgagaac aacggatgga gaaaggtact gataggatat aaacctaatt acctcgagag
     3301 gggacggaaa ctggaaggac aggagcaacc tgtaatgcga atagatataa acctaattac
     3361 ctcgagaggg gacggaaacg gctgttacta aactaatcgt gtatttatgg agatataaac
     3421 ctaattacct cgagagggga cggaaactga acaaagctcc tagaagtctg tgtcgatctg
     3481 atataaacct aattacctcg agaggggacg gaaaccattt aaacagagtg taaattcctt
     3541 ccactcagat ataaacctaa ttacctcgag aggggacgga aacctctgta gaattttgtt
     3601 taagaatact accaagatat aaacctaatt acctcgagag gggacggaaa cctcagggtt
     3661 ttgagaaagg ggacagaatt ggtgatataa acctaattac ctcgagaggg gacggaaaca
     3721 ggttaacact gcccttgaag ctagcagatc tttttccctc tgccaaaaat tatggggaca
     3781 tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa
     3841 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt
     3901 taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg
     3961 ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc
     4021 attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg
     4081 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt
     4141 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg
     4201 acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4261 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4321 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4381 aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa
     4441 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4501 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4561 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc
     4621 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4681 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg
     4741 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4801 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4861 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4921 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4981 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5041 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5101 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5161 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5221 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5281 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5341 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5401 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5461 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5521 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5581 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5641 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5701 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5761 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5821 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5881 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5941 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6001 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6061 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6121 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6181 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6241 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6301 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6361 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6421 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6481 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6541 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6601 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6661 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6721 ccacctg
//
