LOCUS       sequence-268572-        8189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG promoter expression plasmid for human codon optimized NmeCas9 
            nuclease with C-terminal NLS (SV40).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8189)
  AUTHORS   Mahendra C, Christie KA, Osuna BA, Pinilla-Redondo R, Kleinstiver 
            BP, Bondy-Denomy J
  TITLE     Broad-spectrum anti-CRISPR proteins facilitate horizontal gene 
            transfer.
  JOURNAL   Nat Microbiol. 2020 Apr;5(4):620-629. doi: 
            10.1038/s41564-020-0692-2. Epub 2020 Mar 26.
  PUBMED    32218510
REFERENCE   2  (bases 1 to 8189)
  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: 
            "10.1038/s41564-020-0692-2"; journalName: "Nat Microbiol"; date: 
            "2020-04"; volume: "5"; issue: "4"; pages: "620-629"
COMMENT     Sequence Label: pCAG-hNmeCas9-NLS(SV40)-3xFLAG  (KAC409)
FEATURES             Location/Qualifiers
     source          1..8189
                     /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"
     regulatory      1771..1780
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1777..5022
                     /codon_start=1
                     /product="Cas9 endonuclease from the Neisseria meningitidis
                     Type II CRISPR/Cas system (Esvelt et al., 2013)"
                     /label=NmCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MAAFKPNSINYILGLDIGIASVGWAMVEIDEEENPIRLIDLGVRV
                     FERAEVPKTGDSLAMARRLARSVRRLTRRRAHRLLRTRRLLKREGVLQAANFDENGLIK
                     SLPNTPWQLRAAALDRKLTPLEWSAVLLHLIKHRGYLSQRKNEGETADKELGALLKGVA
                     GNAHALQTGDFRTPAELALNKFEKESGHIRNQRSDYSHTFSRKDLQAELILLFEKQKEF
                     GNPHVSGGLKEGIETLLMTQRPALSGDAVQKMLGHCTFEPAEPKAAKNTYTAERFIWLT
                     KLNNLRILEQGSERPLTDTERATLMDEPYRKSKLTYAQARKLLGLEDTAFFKGLRYGKD
                     NAEASTLMEMKAYHAISRALEKEGLKDKKSPLNLSPELQDEIGTAFSLFKTDEDITGRL
                     KDRIQPEILEALLKHISFDKFVQISLKALRRIVPLMEQGKRYDEACAEIYGDHYGKKNT
                     EEKIYLPPIPADEIRNPVVLRALSQARKVINGVVRRYGSPARIHIETAREVGKSFKDRK
                     EIEKRQEENRKDREKAAAKFREYFPNFVGEPKSKDILKLRLYEQQHGKCLYSGKEINLG
                     RLNEKGYVEIDHALPFSRTWDDSFNNKVLVLGSENQNKGNQTPYEYFNGKDNSREWQEF
                     KARVETSRFPRSKKQRILLQKFDEDGFKERNLNDTRYVNRFLCQFVADRMRLTGKGKKR
                     VFASNGQITNLLRGFWGLRKVRAENDRHHALDAVVVACSTVAMQQKITRFVRYKEMNAF
                     DGKTIDKETGEVLHQKTHFPQPWEFFAQEVMIRVFGKPDGKPEFEEADTLEKLRTLLAE
                     KLSSRPEAVHEYVTPLFVSRAPNRKMSGQGHMETVKSAKRLDEGVSVLRVPLTQLKLKD
                     LEKMVNREREPKLYEALKARLEAHKDDPAKAFAEPFYKYDKAGNRTQQVKAVRVEQVQK
                     TGVWVRNHNGIADNATMVRVDVFEKGDKYYLVPIYSWQVAKGILPDRAVVQGKDEEDWQ
                     LIDDSFNFKFSLHPNDLVEVITKKARMFGYFASCHRGTGNINIRIHDLDHKIGKNGILE
                     GIGVKTALSFQKYQIDELGKEIRPCRLKKRPPVR"
     CDS             5029..5049
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5056..5121
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(5211..5230)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5276..5331
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5330..5349)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5692..5708)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5692..5708)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5705..5727)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5716..5732
                     /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(5740..5746,5747..5764,5765..5770))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5785..5806
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5865..6060
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5911..6046
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6066..6200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6103..6122)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6157..6176
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6268..6285)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6439..7027)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6519..6538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7198..7989,7990..8058))
                     /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     7821..7840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8059..8163)
                     /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 gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg 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 ggggccgcct cgggccgggg agggctcggg ggaggggcgc 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 tctgcagtcg
     1741 acggtaccgc gggcccggga tccaccggtc gccaccatgg ctgccttcaa acctaattca
     1801 atcaactaca tcctcggcct cgatatcggc atcgcatccg tcggctgggc gatggtagaa
     1861 attgacgaag aagaaaaccc catccgcctg attgatttgg gcgtgcgcgt atttgagcgt
     1921 gccgaagtac cgaaaacagg cgactccctt gccatggcaa ggcgtttggc gcgcagtgtt
     1981 cgccgcctga cccgccgtcg cgcccaccgc ctgcttcgga cccgccgcct attgaaacgc
     2041 gaaggcgtat tacaagccgc caattttgac gaaaacggct tgattaaatc cttaccgaat
     2101 acaccatggc aacttcgcgc agccgcatta gaccgcaaac tgacgccttt agagtggtcg
     2161 gcagtcttgt tgcatttaat caaacatcgc ggctatttat cgcaacggaa aaacgagggc
     2221 gaaactgccg ataaggagct tggcgctttg cttaaaggcg tagccggcaa tgcccatgcc
     2281 ttacagacag gcgatttccg cacaccggcc gaattggctt taaataaatt tgagaaagaa
     2341 agcggccata tccgcaatca gcgcagcgat tattcgcata cgttcagccg caaagattta
     2401 caggcggagc tgattttgct gtttgaaaaa caaaaagaat ttggcaatcc gcatgtttca
     2461 ggcggcctta aagaaggtat tgaaacccta ctgatgacgc aacgccctgc cctgtccggc
     2521 gatgccgttc aaaaaatgtt ggggcattgc accttcgaac cggcagagcc gaaagccgct
     2581 aaaaacacct acacagccga acgtttcatc tggctgacca agctgaacaa cctgcgtatt
     2641 ttagagcaag gcagcgagcg gccattgacc gataccgaac gcgccacgct tatggacgag
     2701 ccatacagaa aatccaaact gacttacgca caagcccgta agctgctggg tttagaagat
     2761 accgcctttt tcaaaggctt gcgctatggt aaagacaatg ccgaagcctc aacattgatg
     2821 gaaatgaagg cctaccatgc catcagccgt gcactggaaa aagaaggatt gaaagacaaa
     2881 aaatccccat taaacctttc tcccgaatta caagacgaaa tcggcacggc attctccctg
     2941 ttcaaaaccg atgaagacat tacaggccgt ctgaaagacc gtatacagcc cgaaatctta
     3001 gaagcgctgt tgaaacacat cagcttcgat aagttcgtcc aaatttcctt gaaagcattg
     3061 cgccgaattg tgcctctaat ggaacaaggc aaacgttacg atgaagcctg cgccgaaatc
     3121 tacggagacc attacggcaa gaagaatacg gaagaaaaga tttatctgcc gccgattccc
     3181 gccgacgaaa tccgcaaccc cgtcgtcttg cgcgccttat ctcaagcacg taaggtcatt
     3241 aacggcgtgg tacgccgtta cggctcccca gctcgtatcc atattgaaac tgcaagggaa
     3301 gtaggtaaat cgtttaaaga ccgcaaagaa attgagaaac gccaagaaga aaaccgcaaa
     3361 gaccgggaaa aagccgccgc caaattccga gagtatttcc ccaattttgt cggagaaccc
     3421 aaatccaaag atattctgaa actgcgcctg tacgagcaac aacacggcaa atgcctgtat
     3481 tcgggcaaag aaatcaactt aggccgtctg aacgaaaaag gctatgtcga aatcgaccat
     3541 gccctgccgt tctcgcgcac atgggacgac agtttcaaca ataaagtact ggtattgggc
     3601 agcgaaaacc aaaacaaagg caatcaaacc ccttacgaat acttcaacgg caaagacaac
     3661 agccgcgaat ggcaggaatt taaagcgcgt gtcgaaacca gccgtttccc gcgcagtaaa
     3721 aaacaacgga ttctgctgca aaaattcgat gaagacggct ttaaagaacg caatctgaac
     3781 gacacgcgct acgtcaaccg tttcctgtgt caatttgttg ccgaccgtat gcggctgaca
     3841 ggtaaaggca agaaacgtgt ctttgcatcc aacggacaaa ttaccaatct gttgcgcggc
     3901 ttttggggat tgcgcaaagt gcgtgcggaa aacgaccgcc atcacgcctt ggacgccgtc
     3961 gtcgttgcct gctcgaccgt tgccatgcag cagaaaatta cccgttttgt acgctataaa
     4021 gagatgaacg cgtttgacgg taaaaccata gacaaagaaa caggagaagt gctgcatcaa
     4081 aaaacacact tcccacaacc ttgggaattt ttcgcacaag aagtcatgat tcgcgtcttc
     4141 ggcaaaccgg acggcaaacc cgaattcgaa gaagccgata ccctagaaaa actgcgcacg
     4201 ttgcttgccg aaaaattatc atctcgcccc gaagccgtac acgaatacgt tacgccactg
     4261 tttgtttcac gcgcgcccaa tcggaagatg agcgggcaag ggcatatgga gaccgtcaaa
     4321 tccgccaaac gactggacga aggcgtcagc gtgttgcgcg taccgctgac acagttaaaa
     4381 ctgaaagact tggaaaaaat ggtcaatcgg gagcgcgaac ctaagctata cgaagcactg
     4441 aaagcacggc tggaagcaca taaagacgat cctgccaaag cctttgccga gccgttttac
     4501 aaatacgata aagcaggcaa ccgcacccaa caggtaaaag ccgtacgcgt agagcaagta
     4561 cagaaaaccg gcgtatgggt gcgcaaccat aacggtattg ccgacaacgc aaccatggtg
     4621 cgcgtagatg tgtttgagaa aggcgacaag tattatctgg taccgattta cagttggcag
     4681 gtagcgaaag ggattttgcc ggatagggct gttgtacaag gaaaagatga agaagattgg
     4741 caacttattg atgatagttt caactttaaa ttctcattac accctaatga tttagtcgag
     4801 gttataacaa aaaaagctag aatgtttggt tactttgcca gctgccatcg aggcacaggt
     4861 aatatcaata tacgcattca tgatcttgat cataaaattg gcaaaaatgg aatactggaa
     4921 ggtatcggcg tcaaaaccgc cctttcattc caaaaatacc aaattgacga actgggcaaa
     4981 gaaatcagac catgccgtct gaaaaaacgc ccgcctgtcc gtggatcccc caagaagaag
     5041 aggaaagtct cgagcgacta caaagaccat gacggtgatt ataaagatca tgacatcgat
     5101 tacaaggatg acgatgacaa gtaaagcggc cgcactcctc aggtgcaggc tgcctatcag
     5161 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc
     5221 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
     5281 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     5341 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     5401 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat
     5461 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
     5521 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
     5581 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
     5641 cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct
     5701 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5761 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5821 actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca
     5881 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5941 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     6001 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     6061 aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6121 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     6181 tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6241 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6301 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6361 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6421 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6481 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6541 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6601 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6661 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6721 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6781 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6841 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6901 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6961 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7021 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7081 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7141 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7201 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7261 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7321 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7381 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7441 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7501 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7561 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7621 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7681 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7741 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7801 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7861 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7921 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7981 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8041 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8101 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8161 gcgcacattt ccccgaaaag tgccacctg
//
