LOCUS       px458_QC_with_an        9406 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          475..1654
                     /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          470..474
                     /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..427
                     /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          428..433
                     /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        268..343
                     /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        475..760
                     /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    479..748
                     /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        762..1039
                     /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          1040..1268
                     /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(1265..1654)
                     /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             1286..1654
                     /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             1289..1354
                     /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    1289..1309
                     /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    1310..1330
                     /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    1331..1354
                     /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    1361..1384
                     /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             1361..1381
                     /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    1361..1381
                     /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    1367..1381
                     /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             1406..1654
                     /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             5589..5636
                     /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             5652..5705
                     /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             5706..6419
                     /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            5706..6419
                     /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    5706..6419
                     /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(5748..5769)
                     /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    5886..5915
                     /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    6356..6377
                     /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(6447..6464)
                     /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      6450..6653
                     /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    6453..6660
                     /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   6669..6809
                     /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      6884..7339
                     /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      6901..7207
                     /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    7475..7497
                     /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        7621..7725
                     /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        7656..7684
                     /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            7726..8586
                     /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             7726..8586
                     /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      8741..9360
                     /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      8757..9345
                     /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          1655..3256
                     /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(1655..3256)
                     /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             1655..3256
                     /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             1655..3256
                     /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          3257..3538
                     /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(3257..3538)
                     /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             3257..3538
                     /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             3257..3538
                     /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          3621..9406
                     /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(3621..4004)
                     /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             3621..6428
                     /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             3621..5588
                     /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"
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 ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag
      301 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct
      361 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac
      421 aaatggctct agaacgcgtA GGCCTtGAGA CCtagGGTCT Ctgctagcgg tacccgttac
      481 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      541 aatagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      601 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      661 tgacggtaaa tggcccgcct ggcattgtgc ccagtacatg accttatggg actttcctac
      721 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt
      781 ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt
      841 ttaattattt tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc caggcggggc
      901 ggggcggggc gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag
      961 cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa
     1021 gcgaagcgcg cggcgggcgg gagtcgctgc gacgctgcct tcgccccgtg ccccgctccg
     1081 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg
     1141 ggcgggacgg cccttctcct ccgggctgta attagctgag caagaggtaa gggtttaagg
     1201 gatggttggt tggtggggta ttaatgttta attacctgga gcacctgcct gaaatcactt
     1261 tttttcaggt tggACCGGTg ccaccatgga ctataaggac cacgacggag actacaagga
     1321 tcatgatatt gattacaaag acgatgacga taagatggcc ccaaagaaga agcggaaggt
     1381 cggtatccac ggagtcccag cagccgacaa gaagtacagc atcggcctgg acatcggcac
     1441 caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa
     1501 ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt
     1561 cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac
     1621 cagacggaag aaccggatct gctatctgca agAGATCTtc agcaacgaga tggccaaggt
     1681 ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca
     1741 cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc
     1801 caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct
     1861 gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga
     1921 cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa
     1981 ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc
     2041 tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa
     2101 gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa
     2161 gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga
     2221 cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc
     2281 caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac
     2341 caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac
     2401 cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga
     2461 ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta
     2521 caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct
     2581 gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca
     2641 gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct
     2701 gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg
     2761 ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat
     2821 caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga
     2881 gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct
     2941 gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg
     3001 aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt
     3061 caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga
     3121 gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac
     3181 ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga
     3241 ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga
     3301 ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg
     3361 gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa
     3421 gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt
     3481 catgcagctg atCCACGACG ACAGCCTGAC CTTTAAAGAG GACATCCAGA AAGCCCAGGT
     3541 AAGGGGCTCA CAGTAGCAGG CTTGAGGTCT GGACATATAT ATGGGTGACA ATGACATCCA
     3601 CTTTGCCTTT CTCTCCACAG gtgtccggcc agggcgatag cctgcacgag cacattgcca
     3661 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg
     3721 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag
     3781 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag
     3841 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc
     3901 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc
     3961 aggaactgga catcaaccgg ctgtccGACt acgatgtgga ccatatcgtg cctcagagct
     4021 ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca
     4081 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc
     4141 tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag
     4201 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc
     4261 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga
     4321 atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
     4381 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg
     4441 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa
     4501 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg
     4561 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt
     4621 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa
     4681 acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag
     4741 tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca
     4801 gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact
     4861 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg
     4921 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga
     4981 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg
     5041 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc
     5101 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac
     5161 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga
     5221 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc
     5281 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta
     5341 atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg
     5401 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt
     5461 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca
     5521 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg
     5581 gaggcgacaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag aaaaaggaat
     5641 tcggcagtgg agagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg
     5701 gcccagtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     5761 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     5821 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     5881 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     5941 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     6001 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     6061 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     6121 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     6181 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     6241 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     6301 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     6361 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg
     6421 aattctaact agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     6481 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     6541 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     6601 tggggtgggg caggacagca agggggagga ttgggaagag aatagcaggc atgctgggga
     6661 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     6721 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     6781 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     6841 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     6901 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     6961 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     7021 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     7081 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     7141 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     7201 ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     7261 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     7321 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     7381 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     7441 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     7501 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     7561 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     7621 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     7681 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     7741 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     7801 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     7861 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     7921 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     7981 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     8041 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     8101 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     8161 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     8221 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     8281 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     8341 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     8401 ggctggttta ttgctgataa atctggagcc ggtgagcgtg gaagccgcgg tatcattgca
     8461 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     8521 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     8581 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     8641 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     8701 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     8761 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     8821 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     8881 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     8941 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     9001 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     9061 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     9121 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     9181 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     9241 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     9301 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     9361 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt 
//
