LOCUS       sequence-275689-        8430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A human codon-optimized SpCas9 nickase and chimeric guide RNA 
            expression plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8430)
  AUTHORS   Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu X, 
            Jiang W, Marraffini LA, Zhang F
  TITLE     Multiplex Genome Engineering Using CRISPR/Cas Systems.
  JOURNAL   Science. 2013 Jan 3.
  PUBMED    23287718
REFERENCE   2  (bases 1 to 8430)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science. 
            2013 Jan 3."
COMMENT     Sequence Label: pX335-U6-Chimeric_BB-CBh-hSpCas9n(D10A)
FEATURES             Location/Qualifiers
     source          1..8430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        120..405
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     misc_RNA        complement(502..577)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(596..844)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(654..673)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(824..844)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(906..1494)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(986..1005)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1665..2456,2457..2525))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2288..2307
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2526..2630)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2698..2716
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2754..2776)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2876..2895
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2912..3367)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3049..3070)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3261..3280
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3442..3582
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3453..3582
                     /label=AAV2 ITR
     polyA_signal    3591..3798
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3787..3804
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3841..3861)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(3865..7938,7939..7941,7942..7965))
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(3865..7965)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(7975..7995)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(7999..8025)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          8046..8273
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     promoter        complement(join(8274..8430,1..118))
                     /label=chicken beta-actin promoter
ORIGIN
        1 cccccccccc ccgcccccat cgctgcacaa aataattaaa aaataaataa atacaaaatt
       61 gggggtgggg agggggggga gatggggaga gtgaagcaga acgtggggct cacctcgacc
      121 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
      181 atgtactggg cacaatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
      241 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
      301 cattgacgtc aatggaaagt ccctattggc gttactattg acgtcaatgg gcgggggtcg
      361 ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgggtac ctctagagcc
      421 atttgtctgc agaattggcg cacgcgctaa aaacggacta gccttatttt aacttgctat
      481 ttctagctct aaaacaaaaa agcaccgact cggtgccact ttttcaagtt gataacggac
      541 tagccttatt ttaacttgct atttctagct ctaaaacagg tcttctcgaa gacccggtgt
      601 ttcgtccttt ccacaagata tataaagcca agaaatcgaa atactttcaa gttacggtaa
      661 gcatatgata gtccatttta aaacataatt ttaaaactgc aaactaccca agaaattatt
      721 actttctacg tcacgtattt tgtactaata tctttgtgtt tacagtcaaa ttaattccaa
      781 ttatctctct aacagccttg tatcgtatat gcaaatatga aggaatcatg ggaaataggc
      841 cctcacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
      901 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
      961 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1021 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1081 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1141 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1201 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     1261 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     1321 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     1381 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     1441 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     1501 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     1561 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     1621 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     1681 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     1741 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     1801 ccgcggcttc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     1861 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     1921 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     1981 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     2041 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     2101 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     2161 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     2221 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     2281 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     2341 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     2401 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     2461 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     2521 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     2581 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     2641 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     2701 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     2761 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     2821 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     2881 tcagagcaga ttgtactgag agtgcaccat aaaattgtaa acgttaatat tttgttaaaa
     2941 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aatagaccga aatcggcaaa
     3001 atcccttata aatcaaaaga atagcccgag atagagttga gtgttgttcc agtttggaac
     3061 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     3121 ggcgatggcc cactacgtga accatcaccc aaatcaagtt ttttggggtc gaggtgccgt
     3181 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     3241 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctaa ggcgctggca
     3301 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     3361 ggcgcgtact atggttgctt tgacgtatgc ggtgtgaaat accgcacaga tgcgtaagga
     3421 gaaaataccg catcaggcgc ccctgcaggc agctgcgcgc tcgctcgctc actgaggccg
     3481 cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag
     3541 cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc tccccagcat
     3601 gcctgctatt ctcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa
     3661 tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag
     3721 tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct
     3781 ggcaactaga aggcacagtc gaggctgatc agcgagctct aggaattctt agctggcctc
     3841 cacctttctc ttcttcttgg ggctgtcgcc tcccagctga gacaggtcga tccgtgtctc
     3901 gtacaggccg gtgatgctct ggtggatcag ggtggcgtcc agcacctctt tggtgctggt
     3961 gtacctcttc cggtcgatgg tggtgtcaaa gtacttgaag gcggcagggg ctcccagatt
     4021 ggtcagggta aacaggtgga tgatattctc ggcctgctct ctgatgggct tatcccggtg
     4081 cttgttgtag gcggacagca ctttgtccag attagcgtcg gccaggatca ctctcttgga
     4141 gaactcgctg atctgctcga tgatctcgtc caggtagtgc ttgtgctgtt ccacaaacag
     4201 ctgtttctgc tcattatcct cgggggagcc cttcagcttc tcatagtggc tggccaggta
     4261 caggaagttc acatatttgg agggcagggc cagttcgttt cccttctgca gttcgccggc
     4321 agaggccagc attctcttcc ggccgttttc cagctcgaac agggagtact taggcagctt
     4381 gatgatcagg tcctttttca cttctttgta gcccttggct tccagaaagt cgatgggatt
     4441 cttctcgaag ctgcttcttt ccatgatggt gatccccagc agctctttca cactcttcag
     4501 tttcttggac ttgccctttt ccactttggc caccaccagc acagaatagg ccacggtggg
     4561 gctgtcgaag ccgccgtact tcttagggtc ccagtccttc tttctggcga tcagcttatc
     4621 gctgttcctc ttgggcagga tagactcttt gctgaagccg cctgtctgca cctcggtctt
     4681 tttcacgata ttcacttggg gcatgctcag cactttccgc acggtggcaa aatcccggcc
     4741 cttatcccac acgatctccc cggtttcgcc gtttgtctcg atcagaggcc gcttccggat
     4801 ctcgccgttg gccagggtaa tctcggtctt gaaaaagttc atgatgttgc tgtagaagaa
     4861 gtacttggcg gtagccttgc cgatttcctg ctcgctcttg gcgatcatct tccgcacgtc
     4921 gtacaccttg tagtcgccgt acacgaactc gctttccagc ttagggtact ttttgatcag
     4981 ggcggttccc acgacggcgt tcaggtaggc gtcgtgggcg tggtggtagt tgttgatctc
     5041 gcgcactttg taaaactgga aatccttccg gaaatcggac accagcttgg acttcagggt
     5101 gatcactttc acttcccgga tcagcttgtc attctcgtcg tacttagtgt tcatccggga
     5161 gtccaggatc tgtgccacgt gctttgtgat ctgccgggtt tccaccagct gtctcttgat
     5221 gaagccggcc ttatccagtt cgctcaggcc gcctctctcg gccttggtca gattgtcgaa
     5281 ctttctctgg gtaatcagct tggcgttcag cagctgccgc cagtagttct tcatcttctt
     5341 cacgacctct tcggagggca cgttgtcgct cttgccccgg ttcttgtcgc ttctggtcag
     5401 caccttgttg tcgatggagt cgtccttcag aaagctctga ggcacgatat ggtccacatc
     5461 gtagtcggac agccggttga tgtccagttc ctggtccacg tacatatccc gcccattctg
     5521 caggtagtac aggtacagct tctcgttctg cagctgggtg ttttccacgg ggtgttcttt
     5581 caggatctgg ctgcccagct ctttgatgcc ctcttcgatc cgcttcattc tctcgcggct
     5641 gttcttctgt cccttctggg tggtctggtt ctctctggcc atttcgatca cgatgttctc
     5701 gggcttgtgc cggcccatca ctttcacgag ctcgtccacc accttcactg tctgcaggat
     5761 gcccttctta atggcggggc tgccggccag attggcaatg tgctcgtgca ggctatcgcc
     5821 ctggccggac acctgggctt tctggatgtc ctctttaaag gtcaggctgt cgtcgtggat
     5881 cagctgcatg aagtttctgt tggcgaagcc gtcggacttc aggaaatcca ggattgtctt
     5941 gccggactgc ttgtcccgga tgccgttgat cagcttccgg ctcagcctgc cccagccggt
     6001 gtatctccgc cgcttcagct gcttcatcac tttgtcgtcg aacaggtggg cataggtttt
     6061 cagccgttcc tcgatcatct ctctgtcctc aaacagtgtc agggtcagca cgatatcttc
     6121 cagaatgtcc tcgttttcct cattgtccag gaagtccttg tccttgataa ttttcagcag
     6181 atcgtggtat gtgcccaggg aggcgttgaa ccgatcttcc acgccggaga tttccacgga
     6241 gtcgaagcac tcgattttct tgaagtagtc ctctttcagc tgcttcacgg tcactttccg
     6301 gttggtcttg aacagcaggt ccacgatggc ctttttctgc tcgccgctca ggaaggcggg
     6361 ctttctcatt ccctcggtca cgtatttcac tttggtcagc tcgttataca cggtgaagta
     6421 ctcgtacagc aggctgtgct tgggcagcac cttctcgttg ggcaggttct tatcgaagtt
     6481 ggtcatccgc tcgatgaagc tctgggcgga agcgcccttg tccaccactt cctcgaagtt
     6541 ccagggggtg atggtttcct cgctctttct ggtcatccag gcgaatctgc tgtttcccct
     6601 ggccagaggg cccacgtagt aggggatgcg gaaggtcagg atcttctcga tcttttcccg
     6661 gttgtccttc aggaatgggt aaaaatcttc ctgccgccgc agaatggcgt gcagctctcc
     6721 caggtggatc tggtggggga tgctgccgtt gtcgaaggtc cgctgcttcc gcagcaggtc
     6781 ctctctgttc agcttcacga gcagttcctc ggtgccgtcc atcttttcca ggatgggctt
     6841 gatgaacttg tagaactctt cctggctggc tccgccgtca atgtagccgg cgtagccgtt
     6901 cttgctctgg tcgaagaaaa tctctttgta cttctcaggc agctgctgcc gcacgagagc
     6961 tttcagcagg gtcaggtcct ggtggtgctc gtcgtatctc ttgatcatag aggcgctcag
     7021 gggggccttg gtgatctcgg tgttcactct caggatgtcg ctcagcagga tggcgtcgga
     7081 caggttcttg gcggccagaa acaggtcggc gtactggtcg ccgatctggg ccagcaggtt
     7141 gtccaggtcg tcgtcgtagg tgtccttgct cagctgcagt ttggcatcct cggccaggtc
     7201 gaagttgctc ttgaagttgg gggtcaggcc caggctcagg gcaatcaggt tgccgaacag
     7261 gccattcttc ttctcgccgg gcagctgggc gatcagattt tccagccgtc tgctcttgct
     7321 cagtctggca gacaggatgg ccttggcgtc cacgccgctg gcgttgatgg ggttttcctc
     7381 gaacagctgg ttgtaggtct gcaccagctg gatgaacagc ttgtccacgt cgctgttgtc
     7441 ggggttcagg tcgccctcga tcaggaagtg gccccggaac ttgatcatgt gggccagggc
     7501 cagatagatc agccgcaggt cggccttgtc ggtgctgtcc accagtttct ttctcaggtg
     7561 gtagatggtg gggtacttct cgtggtaggc cacctcgtcc acgatgttgc cgaagatggg
     7621 gtgccgctcg tgcttcttat cctcttccac caggaaggac tcttccagtc tgtggaagaa
     7681 gctgtcgtcc accttggcca tctcgttgct gaagatctct tgcagatagc agatccggtt
     7741 cttccgtctg gtgtatcttc ttctggcggt tctcttcagc cgggtggcct cggctgtttc
     7801 gccgctgtcg aacagcaggg ctccgatcag gttcttcttg atgctgtgcc ggtcggtgtt
     7861 gcccagcacc ttgaatttct tgctgggcac cttgtactcg tcggtgatca cggcccagcc
     7921 cacagagttg gtgccgatgg ccaggccgat gctgtacttc ttgtcggacg cttcgacctt
     7981 gcgctttttc ttcggcgaag cgtaatctgg aacatcgtat gggtacatgg tggcaccggt
     8041 ccaacctgaa aaaaagtgat ttcaggcagg tgctccaggt aattaaacat taatacccca
     8101 ccaaccaacc atcccttaaa cccttacctc ttgctcagct aattacagcc cggaggagaa
     8161 gggccgtccc gcccgctcac ctgtgggagt aacgcggtca gtcagagccg gggcgggcgg
     8221 cgcgaggcgg cggcggagcg gggcacgggg cgaaggcagc gcgcagcgac tcccgcccgc
     8281 cgcgcgcttc gctttttata gggccgccgc cgccgccgcc tcgccataaa aggaaacttt
     8341 cggagcgcgc cgctctgatt ggctgccgcc gcacctctcc gcctcgcccc gccccgcccc
     8401 tcgccccgcc ccgccccgcc tggcgcgcgc
//
