LOCUS       pQCi_mB2M_2a_gui        9415 bp    DNA        circular     22-MAY-2020
DEFINITION  pSpCas9(BB)-2A-GFP (PX458).
ACCESSION   .
VERSION     .
KEYWORDS    px458
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9289)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported 18 Feb 2016 from SnapGene Viewer 3.0.3
            http://www.snapgene.com
COMMENT     
COMMENT     
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     source          484..1663
                     /organism="pSpCas9(BB)-2A-GFP (PX458)"
                     /mol_type="other DNA"
                     /note="source:pSpCas9(BB)-2A-GFP (PX458)"
                     /locus_tag="source:pSpCas9(BB)-2A-GFP (PX458)"
                     /label="source:pSpCas9(BB)-2A-GFP (PX458)"
                     /ApEinfo_label="source:pSpCas9(BB)-2A-GFP (PX458)"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     source          479..483
                     /organism="pSpCas9(BB)-2A-GFP (PX458)"
                     /mol_type="other DNA"
                     /note="source:pSpCas9(BB)-2A-GFP (PX458)"
                     /locus_tag="source:pSpCas9(BB)-2A-GFP (PX458)(1)"
                     /label="source:pSpCas9(BB)-2A-GFP (PX458)(1)"
                     /ApEinfo_label="source:pSpCas9(BB)-2A-GFP (PX458)"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     source          1..245
                     /organism="pSpCas9(BB)-2A-GFP (PX458)"
                     /mol_type="other DNA"
                     /note="source:pSpCas9(BB)-2A-GFP (PX458)"
                     /locus_tag="source:pSpCas9(BB)-2A-GFP (PX458)(2)"
                     /label="source:pSpCas9(BB)-2A-GFP (PX458)(2)"
                     /ApEinfo_label="source:pSpCas9(BB)-2A-GFP (PX458)"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     source          431..436
                     /organism="pSpCas9(BB)-2A-GFP (PX458)"
                     /mol_type="other DNA"
                     /note="source:pSpCas9(BB)-2A-GFP (PX458)"
                     /locus_tag="source:pSpCas9(BB)-2A-GFP (PX458)(3)"
                     /label="source:pSpCas9(BB)-2A-GFP (PX458)(3)"
                     /ApEinfo_label="source:pSpCas9(BB)-2A-GFP (PX458)"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        1..241
                     /note="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /locus_tag="U6 promoter"
                     /label="U6 promoter"
                     /ApEinfo_label="U6 promoter"
                     /ApEinfo_fwdcolor="#346ee0"
                     /ApEinfo_revcolor="#346ee0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        10..242
                     /note="hU6_promoter"
                     /locus_tag="hU6_promoter"
                     /label="hU6_promoter"
                     /ApEinfo_label="hU6_promoter"
                     /ApEinfo_fwdcolor="#346ee0"
                     /ApEinfo_revcolor="#346ee0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    172..191
                     /note="LKO1_5_primer"
                     /locus_tag="LKO1_5_primer"
                     /label="LKO1_5_primer"
                     /ApEinfo_label="LKO1_5_primer"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_RNA        271..346
                     /note="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
                     /locus_tag="gRNA scaffold"
                     /label="gRNA scaffold"
                     /ApEinfo_label="gRNA scaffold"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     enhancer        484..769
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer;
                     contains an 18-bp deletion relative to the standard CMV
                     enhancer"
                     /locus_tag="CMV enhancer"
                     /label="CMV enhancer"
                     /ApEinfo_label="CMV enhancer"
                     /ApEinfo_fwdcolor="#5ac3fa"
                     /ApEinfo_revcolor="#5ac3fa"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    488..757
                     /note="CAG_enhancer"
                     /locus_tag="CAG_enhancer"
                     /label="CAG_enhancer"
                     /ApEinfo_label="CAG_enhancer"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        771..1048
                     /note="chicken beta-actin promoter"
                     /locus_tag="chicken beta-actin promoter"
                     /label="chicken beta-actin promoter"
                     /ApEinfo_label="chicken beta-actin promoter"
                     /ApEinfo_fwdcolor="#346ee0"
                     /ApEinfo_revcolor="#346ee0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     intron          1049..1277
                     /note="hybrid intron"
                     /note="hybrid between chicken beta-actin (CBA) and minute
                     virus of mice (MMV) introns (Gray et al., 2011)"
                     /locus_tag="hybrid intron"
                     /label="hybrid intron"
                     /ApEinfo_label="hybrid intron"
                     /ApEinfo_fwdcolor="#eb606c"
                     /ApEinfo_revcolor="#eb606c"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             complement(1274..1663)
                     /codon_start=1
                     /note="ORF frame 3"
                     /translation="MVHIVVGQPVDVQFLVHVHIPPILQVVQVQLLVLQLGVFHGVFF
                     Q
                     DLAAQLFDALFDPLHSLAAVLLSLLGGLVLSGHFDHDVLGLVPAHHFHELVHHLHCLQ
                     D
                     ALLNGGAAGQIGNVLVQAIALAGHLGFLDVLFKGQAVVVDQLHEVSVGEAVGLQEIQD
                     C
                     LAGLLVPDAVDQLPAQPAPAGVSPPLQLLHHFVVEQVGIGFQPFLDHLSVLKQCQGQH
                     D
                     IFQNVLVFLIVQEVLVLDNFQQIVVCAQGGVEPIFHAGDFHGVEALDFLEVVLFQLLH
                     G
                     HFPVGLEQQVHDGLFLLAAQEGGLSHSLGHVFHFGQLVIHGEVLVQQAVLGQHLLVGQ
                     V
                     LIEVGHPLDEALGGSALVHHFLEVPGGDGFLALSGHPGESAVSPGQRAHVVGDAEGQD
                     L
                     LDLFPVVLQEWVKIFLPPQNGVQLSQVDLVGDAAVVEGPLLPQQVLSVQLHEQFLGAV
                     H
                     LFQDGLDELVELFLAGSAVNVAGVAVLALVEENLFVLLRQLLPHESFQQGQVLVVLVV
                     S
                     LDHRGAQGGLGDLGVHSQDVAQQDGVGQVLGGQKQVGVLVADLGQQVVQVVVVGVLAQ
                     L
                     QFGILGQVEVALEVGGQAQAQGNQVSEQAILLLAGQLGDQIFQPSALAQSGRQDGLGV
                     H
                     AAGVDGVFLEQLVVGLHQLDEQLVHVAVVGVQVALDQEVAPELDHVGQGQIDQPQVGL
                     V
                     GAVHQFLSQVVDGGVLLVVGHLVHDVAEDGVPLVLLILFHQEGLFQSVEEAVVHLGHL
                     V
                     AEDLLQIADPVLPSGVSSSGGSLQPGGLGCFAAVEQQGSDQVLLDAVPVGVAQHLEFL
                     A
                     GHLVLVGDHGPAHRVGADVQADAVLLVGCWDSVDTDLPLLLWGHLIVIVFVINIMILV
                     V SVVVLIVHGGTGPT"
                     /locus_tag="ORF frame 3"
                     /label="ORF frame 3"
                     /ApEinfo_label="ORF frame 3"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             1295..1663
                     /codon_start=1
                     /note="ORF frame 3(1)"
                     /translation="MDYKDHDGDYKDHDIDYKDDDDKMAPKKKRKVGIHGVPAADKKY
                     S
                     IGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRL
                     K
                     RTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIV
                     D
                     EVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVD
                     K
                     LFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNL
                     I
                     ALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAI
                     L
                     LSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGY
                     A
                     GYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGE
                     L
                     HAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWN
                     F
                     EEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRK
                     P
                     AFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYH
                     D
                     LLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRR
                     R
                     YTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQV
                     S
                     GQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQK
                     G
                     QKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDIN
                     R
                     LSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAK
                     L
                     ITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKL
                     I
                     REVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEF
                     V
                     YGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNG
                     E
                     TGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDW
                     D
                     PKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAK
                     G
                     YKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEK
                     L
                     KGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIR
                     E
                     QAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDL
                     S
                     QLGGDKRPAATKKAGQAKKKKEFGSGEGRGSLLTCGDVEENPGPVSKGEELFTGVVPI
                     L
                     VELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSR
                     Y
                     PDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKE
                     D
                     GNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDG
                     P VLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKEF"
                     /locus_tag="ORF frame 3(1)"
                     /label="ORF frame 3(1)"
                     /ApEinfo_label="ORF frame 3(1)"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             1298..1363
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by
                     an enterokinase cleavage site"
                     /note="3xFLAG"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /locus_tag="three tandem FLAG(R) epitope tags, followed by
                     an enterokinase cleavage site"
                     /label="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /ApEinfo_label="three tandem FLAG(R) epitope tags,
                     followed by an enterokinase cleavage site"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1298..1318
                     /note="FLAG"
                     /locus_tag="FLAG"
                     /label="FLAG"
                     /ApEinfo_label="FLAG"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1319..1339
                     /note="FLAG(1)"
                     /locus_tag="FLAG(1)"
                     /label="FLAG(1)"
                     /ApEinfo_label="FLAG(1)"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1340..1363
                     /note="FLAG(2)"
                     /locus_tag="FLAG(2)"
                     /label="FLAG(2)"
                     /ApEinfo_label="FLAG(2)"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1370..1393
                     /note="SV40 NLS"
                     /locus_tag="SV40 NLS"
                     /label="SV40 NLS"
                     /ApEinfo_label="SV40 NLS"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             1370..1390
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T
                     antigen"
                     /note="SV40 NLS"
                     /translation="PKKKRKV"
                     /locus_tag="nuclear localization signal of SV40 large T
                     antigen"
                     /label="nuclear localization signal of SV40 large T
                     antigen"
                     /ApEinfo_label="nuclear localization signal of SV40 large
                     T antigen"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1370..1390
                     /note="SV40 NLS(1)"
                     /locus_tag="SV40 NLS(1)"
                     /label="SV40 NLS(1)"
                     /ApEinfo_label="SV40 NLS(1)"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1376..1390
                     /note="NLS"
                     /locus_tag="NLS"
                     /label="NLS"
                     /ApEinfo_label="NLS"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             1415..1663
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
                     /note="Cas9"
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     N
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     F
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     F
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     E
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     I
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     Q
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     R
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     N
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     F
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     S
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     L
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     Q
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     H
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     L
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     S
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     V
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     N
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     E
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     E
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     T
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     L
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     A
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     D ATLIHQSITGLYETRIDLSQLGGD"
                     /locus_tag="Cas9 (Csn1) endonuclease from the
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
                     /ApEinfo_label="Cas9 (Csn1) endonuclease from the
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             5598..5645
                     /codon_start=1
                     /product="bipartite nuclear localization signal from
                     nucleoplasmin"
                     /note="nucleoplasmin NLS"
                     /translation="KRPAATKKAGQAKKKK"
                     /locus_tag="bipartite nuclear localization signal from
                     nucleoplasmin"
                     /label="bipartite nuclear localization signal from
                     nucleoplasmin"
                     /ApEinfo_label="bipartite nuclear localization signal from
                     nucleoplasmin"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             5661..5714
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /note="T2A"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
                     /locus_tag="2A peptide from Thosea asigna virus capsid
                     protein"
                     /label="2A peptide from Thosea asigna virus capsid
                     protein"
                     /ApEinfo_label="2A peptide from Thosea asigna virus capsid
                     protein"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             5715..6428
                     /codon_start=1
                     /product="enhanced GFP"
                     /note="EGFP"
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     K
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     G
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     V
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     E FVTAAGITLGMDELYK"
                     /locus_tag="enhanced GFP"
                     /label="enhanced GFP"
                     /ApEinfo_label="enhanced GFP"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     gene            5715..6428
                     /gene="EGFP"
                     /note="EGFP"
                     /locus_tag="EGFP"
                     /label="EGFP"
                     /ApEinfo_label="EGFP"
                     /ApEinfo_fwdcolor="#ff797d"
                     /ApEinfo_revcolor="#ff102f"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    5715..6428
                     /note="EGFP(1)"
                     /locus_tag="EGFP(1)"
                     /label="EGFP(1)"
                     /ApEinfo_label="EGFP(1)"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    complement(5757..5778)
                     /note="EGFP_N_primer"
                     /locus_tag="EGFP_N_primer"
                     /label="EGFP_N_primer"
                     /ApEinfo_label="EGFP_N_primer"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    5895..5924
                     /note="Y66 (EGFP)"
                     /locus_tag="Y66 (EGFP)"
                     /label="Y66 (EGFP)"
                     /ApEinfo_label="Y66 (EGFP)"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    6365..6386
                     /note="EGFP_C_primer"
                     /locus_tag="EGFP_C_primer"
                     /label="EGFP_C_primer"
                     /ApEinfo_label="EGFP_C_primer"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    complement(6456..6473)
                     /note="BGH_rev_primer"
                     /locus_tag="BGH_rev_primer"
                     /label="BGH_rev_primer"
                     /ApEinfo_label="BGH_rev_primer"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     terminator      6459..6662
                     /note="bGH_PA_terminator"
                     /locus_tag="bGH_PA_terminator"
                     /label="bGH_PA_terminator"
                     /ApEinfo_label="bGH_PA_terminator"
                     /ApEinfo_fwdcolor="#9d1b1c"
                     /ApEinfo_revcolor="#9d1b1c"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     polyA_signal    6462..6669
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
                     /locus_tag="bGH poly(A) signal"
                     /label="bGH poly(A) signal"
                     /ApEinfo_label="bGH poly(A) signal"
                     /ApEinfo_fwdcolor="#ff3eee"
                     /ApEinfo_revcolor="#ff3eee"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     repeat_region   6678..6818
                     /note="AAV2 ITR"
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
                     /locus_tag="AAV2 ITR"
                     /label="AAV2 ITR"
                     /ApEinfo_label="AAV2 ITR"
                     /ApEinfo_fwdcolor="#0dfff7"
                     /ApEinfo_revcolor="#0dfff7"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     rep_origin      6893..7348
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /locus_tag="f1 ori"
                     /label="f1 ori"
                     /ApEinfo_label="f1 ori"
                     /ApEinfo_fwdcolor="#999999"
                     /ApEinfo_revcolor="#999999"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     rep_origin      6910..7216
                     /note="f1_origin"
                     /locus_tag="f1_origin"
                     /label="f1_origin"
                     /ApEinfo_label="f1_origin"
                     /ApEinfo_fwdcolor="#999999"
                     /ApEinfo_revcolor="#999999"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    7484..7506
                     /note="pGEX_3_primer"
                     /locus_tag="pGEX_3_primer"
                     /label="pGEX_3_primer"
                     /ApEinfo_label="pGEX_3_primer"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        7630..7734
                     /gene="bla"
                     /note="AmpR promoter"
                     /locus_tag="bla"
                     /label="bla"
                     /ApEinfo_label="bla"
                     /ApEinfo_fwdcolor="#346ee0"
                     /ApEinfo_revcolor="#346ee0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        7665..7693
                     /note="AmpR_promoter"
                     /locus_tag="AmpR_promoter"
                     /label="AmpR_promoter"
                     /ApEinfo_label="AmpR_promoter"
                     /ApEinfo_fwdcolor="#346ee0"
                     /ApEinfo_revcolor="#346ee0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     gene            7735..8595
                     /gene="Ampicillin"
                     /note="Ampicillin"
                     /locus_tag="Ampicillin"
                     /label="Ampicillin"
                     /ApEinfo_label="Ampicillin"
                     /ApEinfo_fwdcolor="#ff797d"
                     /ApEinfo_revcolor="#ff102f"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             7735..8595
                     /codon_start=1
                     /note="ORF frame 1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     I
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     S
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     W
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     A
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     S LIKHW"
                     /locus_tag="ORF frame 1"
                     /label="ORF frame 1"
                     /ApEinfo_label="ORF frame 1"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     rep_origin      8750..9369
                     /note="pBR322_origin"
                     /locus_tag="pBR322_origin"
                     /label="pBR322_origin"
                     /ApEinfo_label="pBR322_origin"
                     /ApEinfo_fwdcolor="#999999"
                     /ApEinfo_revcolor="#999999"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     rep_origin      8766..9354
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /locus_tag="ori"
                     /label="ori"
                     /ApEinfo_label="ori"
                     /ApEinfo_fwdcolor="#999999"
                     /ApEinfo_revcolor="#999999"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     source          1664..3265
                     /organism="pSpCas9(BB)-2A-GFP (PX458)"
                     /mol_type="other DNA"
                     /note="source:pSpCas9(BB)-2A-GFP (PX458)"
                     /locus_tag="source:pSpCas9(BB)-2A-GFP (PX458)(4)"
                     /label="source:pSpCas9(BB)-2A-GFP (PX458)(4)"
                     /ApEinfo_label="source:pSpCas9(BB)-2A-GFP (PX458)"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             complement(1664..3265)
                     /codon_start=1
                     /note="ORF frame 3"
                     /translation="MVHIVVGQPVDVQFLVHVHIPPILQVVQVQLLVLQLGVFHGVFF
                     Q
                     DLAAQLFDALFDPLHSLAAVLLSLLGGLVLSGHFDHDVLGLVPAHHFHELVHHLHCLQ
                     D
                     ALLNGGAAGQIGNVLVQAIALAGHLGFLDVLFKGQAVVVDQLHEVSVGEAVGLQEIQD
                     C
                     LAGLLVPDAVDQLPAQPAPAGVSPPLQLLHHFVVEQVGIGFQPFLDHLSVLKQCQGQH
                     D
                     IFQNVLVFLIVQEVLVLDNFQQIVVCAQGGVEPIFHAGDFHGVEALDFLEVVLFQLLH
                     G
                     HFPVGLEQQVHDGLFLLAAQEGGLSHSLGHVFHFGQLVIHGEVLVQQAVLGQHLLVGQ
                     V
                     LIEVGHPLDEALGGSALVHHFLEVPGGDGFLALSGHPGESAVSPGQRAHVVGDAEGQD
                     L
                     LDLFPVVLQEWVKIFLPPQNGVQLSQVDLVGDAAVVEGPLLPQQVLSVQLHEQFLGAV
                     H
                     LFQDGLDELVELFLAGSAVNVAGVAVLALVEENLFVLLRQLLPHESFQQGQVLVVLVV
                     S
                     LDHRGAQGGLGDLGVHSQDVAQQDGVGQVLGGQKQVGVLVADLGQQVVQVVVVGVLAQ
                     L
                     QFGILGQVEVALEVGGQAQAQGNQVSEQAILLLAGQLGDQIFQPSALAQSGRQDGLGV
                     H
                     AAGVDGVFLEQLVVGLHQLDEQLVHVAVVGVQVALDQEVAPELDHVGQGQIDQPQVGL
                     V
                     GAVHQFLSQVVDGGVLLVVGHLVHDVAEDGVPLVLLILFHQEGLFQSVEEAVVHLGHL
                     V
                     AEDLLQIADPVLPSGVSSSGGSLQPGGLGCFAAVEQQGSDQVLLDAVPVGVAQHLEFL
                     A
                     GHLVLVGDHGPAHRVGADVQADAVLLVGCWDSVDTDLPLLLWGHLIVIVFVINIMILV
                     V SVVVLIVHGGTGPT"
                     /locus_tag="ORF frame 3(2)"
                     /label="ORF frame 3(2)"
                     /ApEinfo_label="ORF frame 3"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             1664..3265
                     /codon_start=1
                     /note="ORF frame 3(1)"
                     /translation="MDYKDHDGDYKDHDIDYKDDDDKMAPKKKRKVGIHGVPAADKKY
                     S
                     IGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRL
                     K
                     RTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIV
                     D
                     EVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVD
                     K
                     LFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNL
                     I
                     ALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAI
                     L
                     LSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGY
                     A
                     GYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGE
                     L
                     HAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWN
                     F
                     EEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRK
                     P
                     AFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYH
                     D
                     LLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRR
                     R
                     YTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQV
                     S
                     GQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQK
                     G
                     QKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDIN
                     R
                     LSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAK
                     L
                     ITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKL
                     I
                     REVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEF
                     V
                     YGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNG
                     E
                     TGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDW
                     D
                     PKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAK
                     G
                     YKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEK
                     L
                     KGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIR
                     E
                     QAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDL
                     S
                     QLGGDKRPAATKKAGQAKKKKEFGSGEGRGSLLTCGDVEENPGPVSKGEELFTGVVPI
                     L
                     VELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSR
                     Y
                     PDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKE
                     D
                     GNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDG
                     P VLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKEF"
                     /locus_tag="ORF frame 3(1)(1)"
                     /label="ORF frame 3(1)(1)"
                     /ApEinfo_label="ORF frame 3(1)"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             1664..3265
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
                     /note="Cas9"
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     N
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     F
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     F
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     E
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     I
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     Q
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     R
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     N
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     F
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     S
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     L
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     Q
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     H
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     L
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     S
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     V
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     N
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     E
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     E
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     T
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     L
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     A
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     D ATLIHQSITGLYETRIDLSQLGGD"
                     /locus_tag="Cas9 (Csn1) endonuclease from the
                     Streptococcus pyogenes Type II CRISPR/Cas system(1)"
                     /label="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system(1)"
                     /ApEinfo_label="Cas9 (Csn1) endonuclease from the
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     source          3266..3547
                     /organism="pSpCas9(BB)-2A-GFP (PX458)"
                     /mol_type="other DNA"
                     /note="source:pSpCas9(BB)-2A-GFP (PX458)"
                     /locus_tag="source:pSpCas9(BB)-2A-GFP (PX458)(5)"
                     /label="source:pSpCas9(BB)-2A-GFP (PX458)(5)"
                     /ApEinfo_label="source:pSpCas9(BB)-2A-GFP (PX458)"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             complement(3266..3547)
                     /codon_start=1
                     /note="ORF frame 3"
                     /translation="MVHIVVGQPVDVQFLVHVHIPPILQVVQVQLLVLQLGVFHGVFF
                     Q
                     DLAAQLFDALFDPLHSLAAVLLSLLGGLVLSGHFDHDVLGLVPAHHFHELVHHLHCLQ
                     D
                     ALLNGGAAGQIGNVLVQAIALAGHLGFLDVLFKGQAVVVDQLHEVSVGEAVGLQEIQD
                     C
                     LAGLLVPDAVDQLPAQPAPAGVSPPLQLLHHFVVEQVGIGFQPFLDHLSVLKQCQGQH
                     D
                     IFQNVLVFLIVQEVLVLDNFQQIVVCAQGGVEPIFHAGDFHGVEALDFLEVVLFQLLH
                     G
                     HFPVGLEQQVHDGLFLLAAQEGGLSHSLGHVFHFGQLVIHGEVLVQQAVLGQHLLVGQ
                     V
                     LIEVGHPLDEALGGSALVHHFLEVPGGDGFLALSGHPGESAVSPGQRAHVVGDAEGQD
                     L
                     LDLFPVVLQEWVKIFLPPQNGVQLSQVDLVGDAAVVEGPLLPQQVLSVQLHEQFLGAV
                     H
                     LFQDGLDELVELFLAGSAVNVAGVAVLALVEENLFVLLRQLLPHESFQQGQVLVVLVV
                     S
                     LDHRGAQGGLGDLGVHSQDVAQQDGVGQVLGGQKQVGVLVADLGQQVVQVVVVGVLAQ
                     L
                     QFGILGQVEVALEVGGQAQAQGNQVSEQAILLLAGQLGDQIFQPSALAQSGRQDGLGV
                     H
                     AAGVDGVFLEQLVVGLHQLDEQLVHVAVVGVQVALDQEVAPELDHVGQGQIDQPQVGL
                     V
                     GAVHQFLSQVVDGGVLLVVGHLVHDVAEDGVPLVLLILFHQEGLFQSVEEAVVHLGHL
                     V
                     AEDLLQIADPVLPSGVSSSGGSLQPGGLGCFAAVEQQGSDQVLLDAVPVGVAQHLEFL
                     A
                     GHLVLVGDHGPAHRVGADVQADAVLLVGCWDSVDTDLPLLLWGHLIVIVFVINIMILV
                     V SVVVLIVHGGTGPT"
                     /locus_tag="ORF frame 3(3)"
                     /label="ORF frame 3(3)"
                     /ApEinfo_label="ORF frame 3"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             3266..3547
                     /codon_start=1
                     /note="ORF frame 3(1)"
                     /translation="MDYKDHDGDYKDHDIDYKDDDDKMAPKKKRKVGIHGVPAADKKY
                     S
                     IGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRL
                     K
                     RTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIV
                     D
                     EVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVD
                     K
                     LFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNL
                     I
                     ALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAI
                     L
                     LSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGY
                     A
                     GYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGE
                     L
                     HAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWN
                     F
                     EEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRK
                     P
                     AFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYH
                     D
                     LLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRR
                     R
                     YTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQV
                     S
                     GQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQK
                     G
                     QKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDIN
                     R
                     LSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAK
                     L
                     ITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKL
                     I
                     REVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEF
                     V
                     YGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNG
                     E
                     TGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDW
                     D
                     PKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAK
                     G
                     YKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEK
                     L
                     KGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIR
                     E
                     QAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDL
                     S
                     QLGGDKRPAATKKAGQAKKKKEFGSGEGRGSLLTCGDVEENPGPVSKGEELFTGVVPI
                     L
                     VELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSR
                     Y
                     PDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKE
                     D
                     GNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDG
                     P VLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKEF"
                     /locus_tag="ORF frame 3(1)(2)"
                     /label="ORF frame 3(1)(2)"
                     /ApEinfo_label="ORF frame 3(1)"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             3266..3547
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
                     /note="Cas9"
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     N
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     F
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     F
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     E
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     I
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     Q
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     R
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     N
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     F
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     S
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     L
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     Q
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     H
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     L
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     S
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     V
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     N
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     E
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     E
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     T
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     L
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     A
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     D ATLIHQSITGLYETRIDLSQLGGD"
                     /locus_tag="Cas9 (Csn1) endonuclease from the
                     Streptococcus pyogenes Type II CRISPR/Cas system(2)"
                     /label="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system(2)"
                     /ApEinfo_label="Cas9 (Csn1) endonuclease from the
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     source          3630..9415
                     /organism="pSpCas9(BB)-2A-GFP (PX458)"
                     /mol_type="other DNA"
                     /note="source:pSpCas9(BB)-2A-GFP (PX458)"
                     /locus_tag="source:pSpCas9(BB)-2A-GFP (PX458)(6)"
                     /label="source:pSpCas9(BB)-2A-GFP (PX458)(6)"
                     /ApEinfo_label="source:pSpCas9(BB)-2A-GFP (PX458)"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             complement(3630..4013)
                     /codon_start=1
                     /note="ORF frame 3"
                     /translation="MVHIVVGQPVDVQFLVHVHIPPILQVVQVQLLVLQLGVFHGVFF
                     Q
                     DLAAQLFDALFDPLHSLAAVLLSLLGGLVLSGHFDHDVLGLVPAHHFHELVHHLHCLQ
                     D
                     ALLNGGAAGQIGNVLVQAIALAGHLGFLDVLFKGQAVVVDQLHEVSVGEAVGLQEIQD
                     C
                     LAGLLVPDAVDQLPAQPAPAGVSPPLQLLHHFVVEQVGIGFQPFLDHLSVLKQCQGQH
                     D
                     IFQNVLVFLIVQEVLVLDNFQQIVVCAQGGVEPIFHAGDFHGVEALDFLEVVLFQLLH
                     G
                     HFPVGLEQQVHDGLFLLAAQEGGLSHSLGHVFHFGQLVIHGEVLVQQAVLGQHLLVGQ
                     V
                     LIEVGHPLDEALGGSALVHHFLEVPGGDGFLALSGHPGESAVSPGQRAHVVGDAEGQD
                     L
                     LDLFPVVLQEWVKIFLPPQNGVQLSQVDLVGDAAVVEGPLLPQQVLSVQLHEQFLGAV
                     H
                     LFQDGLDELVELFLAGSAVNVAGVAVLALVEENLFVLLRQLLPHESFQQGQVLVVLVV
                     S
                     LDHRGAQGGLGDLGVHSQDVAQQDGVGQVLGGQKQVGVLVADLGQQVVQVVVVGVLAQ
                     L
                     QFGILGQVEVALEVGGQAQAQGNQVSEQAILLLAGQLGDQIFQPSALAQSGRQDGLGV
                     H
                     AAGVDGVFLEQLVVGLHQLDEQLVHVAVVGVQVALDQEVAPELDHVGQGQIDQPQVGL
                     V
                     GAVHQFLSQVVDGGVLLVVGHLVHDVAEDGVPLVLLILFHQEGLFQSVEEAVVHLGHL
                     V
                     AEDLLQIADPVLPSGVSSSGGSLQPGGLGCFAAVEQQGSDQVLLDAVPVGVAQHLEFL
                     A
                     GHLVLVGDHGPAHRVGADVQADAVLLVGCWDSVDTDLPLLLWGHLIVIVFVINIMILV
                     V SVVVLIVHGGTGPT"
                     /locus_tag="ORF frame 3(4)"
                     /label="ORF frame 3(4)"
                     /ApEinfo_label="ORF frame 3"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             3630..6437
                     /codon_start=1
                     /note="ORF frame 3(1)"
                     /translation="MDYKDHDGDYKDHDIDYKDDDDKMAPKKKRKVGIHGVPAADKKY
                     S
                     IGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRL
                     K
                     RTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIV
                     D
                     EVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVD
                     K
                     LFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNL
                     I
                     ALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAI
                     L
                     LSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGY
                     A
                     GYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGE
                     L
                     HAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWN
                     F
                     EEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRK
                     P
                     AFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYH
                     D
                     LLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRR
                     R
                     YTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQV
                     S
                     GQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQK
                     G
                     QKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDIN
                     R
                     LSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAK
                     L
                     ITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKL
                     I
                     REVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEF
                     V
                     YGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNG
                     E
                     TGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDW
                     D
                     PKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAK
                     G
                     YKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEK
                     L
                     KGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIR
                     E
                     QAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDL
                     S
                     QLGGDKRPAATKKAGQAKKKKEFGSGEGRGSLLTCGDVEENPGPVSKGEELFTGVVPI
                     L
                     VELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSR
                     Y
                     PDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKE
                     D
                     GNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDG
                     P VLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKEF"
                     /locus_tag="ORF frame 3(1)(3)"
                     /label="ORF frame 3(1)(3)"
                     /ApEinfo_label="ORF frame 3(1)"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             3630..5597
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
                     /note="Cas9"
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     N
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     F
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     F
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     E
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     I
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     Q
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     R
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     N
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     F
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     S
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     L
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     Q
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     H
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     L
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     S
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     V
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     N
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     E
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     E
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     T
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     L
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     A
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     D ATLIHQSITGLYETRIDLSQLGGD"
                     /locus_tag="Cas9 (Csn1) endonuclease from the
                     Streptococcus pyogenes Type II CRISPR/Cas system(3)"
                     /label="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system(3)"
                     /ApEinfo_label="Cas9 (Csn1) endonuclease from the
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     source          271..430
                     /organism="pSpCas9(BB)-2A-GFP (PX458)"
                     /mol_type="other DNA"
                     /note="source:pSpCas9(BB)-2A-GFP (PX458)"
                     /locus_tag="source:pSpCas9(BB)-2A-GFP (PX458)(7)"
                     /label="source:pSpCas9(BB)-2A-GFP (PX458)(7)"
                     /ApEinfo_label="source:pSpCas9(BB)-2A-GFP (PX458)"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaaCACCG TCGGCTTCCC ATTCTCCGGT gttttagagc tagaaatagc aagttaaaat
      301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
      361 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
      421 gacaaatggc tctagaacgc gtAGGCCTTC GGCTTCCCAT TCTCCGGTGG Ggctagcggt
      481 acccgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      541 attgacgtca atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      601 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      661 tgacgtcaat gacggtaaat ggcccgcctg gcattgtgcc cagtacatga ccttatggga
      721 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
      781 ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt
      841 atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc
      901 aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc
      961 caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc
     1021 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg acgctgcctt cgccccgtgc
     1081 cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac
     1141 aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagctgagc aagaggtaag
     1201 ggtttaaggg atggttggtt ggtggggtat taatgtttaa ttacctggag cacctgcctg
     1261 aaatcacttt ttttcaggtt ggACCGGTgc caccatggac tataaggacc acgacggaga
     1321 ctacaaggat catgatattg attacaaaga cgatgacgat aagatggccc caaagaagaa
     1381 gcggaaggtc ggtatccacg gagtcccagc agccgacaag aagtacagca tcggcctgga
     1441 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
     1501 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
     1561 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
     1621 aagatacacc agacggaaga accggatctg ctatctgcaa gAGATCTtca gcaacgagat
     1681 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
     1741 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
     1801 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
     1861 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
     1921 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
     1981 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
     2041 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
     2101 cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc
     2161 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
     2221 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
     2281 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
     2341 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
     2401 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
     2461 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
     2521 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
     2581 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
     2641 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
     2701 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
     2761 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
     2821 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
     2881 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
     2941 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
     3001 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
     3061 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
     3121 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
     3181 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
     3241 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
     3301 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
     3361 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
     3421 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
     3481 cagaaacttc atgcagctga tCCACGACGA CAGCCTGACC TTTAAAGAGG ACATCCAGAA
     3541 AGCCCAGGTA AGGGGCTCAC AGTAGCAGGC TTGAGGTCTG GACATATATA TGGGTGACAA
     3601 TGACATCCAC TTTGCCTTTC TCTCCACAGg tgtccggcca gggcgatagc ctgcacgagc
     3661 acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg
     3721 tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa
     3781 tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc
     3841 ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa
     3901 acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt
     3961 acgtggacca ggaactggac atcaaccggc tgtccGACta cgatgtggac catatcgtgc
     4021 ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga
     4081 accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact
     4141 ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg
     4201 ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg
     4261 aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt
     4321 acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg
     4381 tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc
     4441 acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta
     4501 agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg
     4561 ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca
     4621 tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga
     4681 tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg
     4741 tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag
     4801 gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa
     4861 agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg
     4921 tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc
     4981 tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg
     5041 aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc
     5101 tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg
     5161 gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg
     5221 agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca
     5281 agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg
     5341 ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca
     5401 gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg
     5461 ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc
     5521 tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt
     5581 ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga
     5641 aaaaggaatt cggcagtgga gagggcagag gaagtctgct aacatgcggt gacgtcgagg
     5701 agaatcctgg cccagtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     5761 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     5821 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     5881 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     5941 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     6001 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     6061 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     6121 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     6181 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     6241 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     6301 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     6361 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     6421 tgtacaagga attctaacta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
     6481 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     6541 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     6601 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaga atagcaggca
     6661 tgctggggag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     6721 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     6781 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     6841 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     6901 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     6961 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     7021 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     7081 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     7141 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     7201 ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
     7261 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     7321 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     7381 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     7441 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     7501 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     7561 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     7621 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     7681 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     7741 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     7801 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     7861 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     7921 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     7981 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     8041 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     8101 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     8161 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     8221 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     8281 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     8341 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     8401 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg aagccgcggt
     8461 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     8521 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     8581 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     8641 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     8701 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     8761 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     8821 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     8881 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     8941 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     9001 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     9061 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     9121 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     9181 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     9241 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     9301 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     9361 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
