LOCUS       sequence-419586-       13020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing Cas9 fused with GFP at the C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13020)
  AUTHORS   Aoki K, Hyuga M, Tarumoto Y, Nishibuchi G, Ueda A, Ochi Y, Sugino S,
            Mikami T, Kobushi H, Kato I, Akahane K, Inukai T, Takaori-Kondo A, 
            Takita J, Ogawa S, Yusa K
  TITLE     Canonical BAF complex regulates the oncogenic program in human 
            T-cell acute lymphoblastic leukemia.
  JOURNAL   Blood. 2023 Nov 3:blood.2023020857. doi: 10.1182/blood.2023020857.
  PUBMED    37922452
REFERENCE   2  (bases 1 to 13020)
  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: "Blood. 2023
            Nov 3:blood.2023020857. doi: 10.1182/blood.2023020857."
COMMENT     Sequence Label: pKLV2-EF1a-Cas9GFP-W
FEATURES             Location/Qualifiers
     source          1..13020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(22..450)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(132..153)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     344..363
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     577..599
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     591..608
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     592..608
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     618..637
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        618..636
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        952..1150
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1105..1125
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1168..1348
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1395..1520
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2019..2252
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2437..2481
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2779..2896
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3367..4548
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3598..4540
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4497..4517
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      4561..4570
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4570..4635
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4642..4662
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4687..8787
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4687..8787
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             8788..8835
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8854..8907
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(8908..8910,8911..8913,8914..9627)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(8908..8910,8911..8913,8914..9624)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(9214..9233)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9561..9582
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(9687..9707)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10105..10116)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10568..10585)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10574..10798
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(10824..10844)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(10824..10842)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10863..10879)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10863..10879)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10876..10898)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10887..10903
                     /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(10911..10917,10918..10935,10936..10941))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10956..10977
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11094..11111)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11265..11853)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11345..11364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12024..12815,12816..12884))
                     /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     12647..12666
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12885..12989)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
       61 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
      121 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
      181 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
      241 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
      301 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
      361 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
      421 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
      481 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
      541 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
      601 cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgactagt acataactta
      661 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      721 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      781 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      841 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      901 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      961 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     1021 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     1081 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     1141 atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
     1201 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     1261 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     1321 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     1381 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     1441 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     1501 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     1561 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     1621 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     1681 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     1741 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     1801 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     1861 ccggccgcgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
     1921 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
     1981 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
     2041 ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga
     2101 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
     2161 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
     2221 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
     2281 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
     2341 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
     2401 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
     2461 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
     2521 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
     2581 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
     2641 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
     2701 gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat
     2761 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
     2821 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
     2881 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt ggttagtacc
     2941 gggccctacg cgtgaggggc gcggaagccg gggcggagga ggagccaacc ccgggctggg
     3001 ctgagacccg cagaggaaga cgctctaggg atttgtcccg gactagcgag atggcaaggc
     3061 tgaggacggg aggctgattg agaggcgaag gtacacccta atctcaatac aacctttgga
     3121 gctaagccag caatggtaga gggaagattc tgcacgtccc ttccaggcgg cctccccgtc
     3181 accacccccc ccaacccgcc ccgaccggag ctgagagtaa ttcatacaaa aggactcgcc
     3241 cctgccttgg ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat
     3301 cgctgcgttc ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat
     3361 gcgtgaggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     3421 ttggggggag gggtcggcaa ttgaaccggt gcctagagaa agtggcgcgg ggtaaactgg
     3481 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     3541 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta
     3601 agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct
     3661 tgaattactt ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg
     3721 aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt
     3781 tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg
     3841 tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct
     3901 ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt
     3961 ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg
     4021 gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc
     4081 tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg
     4141 gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc
     4201 aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag
     4261 ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag
     4321 gcacctcgat tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt
     4381 ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca
     4441 cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa
     4501 gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtgga agcttggcgc
     4561 gccaccatgg actataagga ccacgacgga gactacaagg atcatgatat tgattacaaa
     4621 gacgatgacg ataagatggc cccaaagaag aagcggaagg tcggtatcca cggagtccca
     4681 gcagccgaca agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc
     4741 gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac
     4801 cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc
     4861 gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc
     4921 tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac
     4981 agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc
     5041 ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga
     5101 aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc
     5161 cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc
     5221 gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac
     5281 cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc
     5341 agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga
     5401 aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc
     5461 gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg
     5521 gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc
     5581 atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc
     5641 tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg
     5701 cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac
     5761 gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc
     5821 ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg
     5881 cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg
     5941 cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag
     6001 atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac
     6061 agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag
     6121 gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat
     6181 aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc
     6241 gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc
     6301 ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg
     6361 accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa
     6421 atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa
     6481 attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc
     6541 gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat
     6601 gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg
     6661 ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc
     6721 ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac
     6781 gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc
     6841 ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag
     6901 acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc
     6961 gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag
     7021 agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac
     7081 cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg
     7141 cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac
     7201 catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga
     7261 agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg
     7321 aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat
     7381 ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga
     7441 cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg
     7501 aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag
     7561 tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac
     7621 aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa
     7681 aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg
     7741 aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac
     7801 agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag
     7861 cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat
     7921 tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag
     7981 gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg
     8041 atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg
     8101 gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt
     8161 gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc
     8221 gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct
     8281 aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa
     8341 ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc
     8401 agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg
     8461 gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga
     8521 gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat
     8581 aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga
     8641 gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc
     8701 aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg
     8761 atcgacctgt ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag
     8821 gcaaaaaaga aaaagggcgg cgggtccgga ggagagggca gaggaagtct cctaacatgc
     8881 ggtgacgtgg aggagaatcc tggcccaatg gtgagcaagg gcgaggagct gttcaccggg
     8941 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     9001 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     9061 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     9121 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     9181 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     9241 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     9301 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     9361 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     9421 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     9481 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     9541 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     9601 ctcggcatgg acgagctgta caagtaagcg gccgctcaac ctctggatta caaaatttgt
     9661 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     9721 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     9781 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     9841 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     9901 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     9961 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
    10021 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
    10081 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
    10141 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
    10201 tccctttggg ccgcctcccc gcaggtacct ttaagaccaa tgacttacaa ggcagctgta
    10261 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaaaga
    10321 agtcaagatc tgctttttgc ctgtactggg tctctctggt tagaccagag tctctctggt
    10381 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
    10441 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
    10501 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ttaaacccgc
    10561 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
    10621 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
    10681 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
    10741 aagggggagg attgggaagt caatagcagg catgctgggg atgcggtggg ctctatgggc
    10801 ggccgttaat taagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
    10861 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
    10921 gccgggagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
    10981 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
    11041 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
    11101 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
    11161 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
    11221 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
    11281 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
    11341 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
    11401 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
    11461 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
    11521 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
    11581 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
    11641 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
    11701 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
    11761 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
    11821 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
    11881 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
    11941 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
    12001 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
    12061 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
    12121 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
    12181 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
    12241 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
    12301 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
    12361 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
    12421 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
    12481 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
    12541 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
    12601 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
    12661 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
    12721 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
    12781 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
    12841 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
    12901 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
    12961 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
//
