pCMV-PE5max-FANCF-U6-PegRNA-H1-Nicking sgRNA

LOCUS       pCMV-PE5max-FANCF-U6-PegRNA-H1-Nicking sgRNA              14884 bp DNA     circular SYN 07-NOV-2025
DEFINITION  expresses sgRNA from the U6 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14884)
  AUTHORS   Shen B, Zhang W, Zhang J, Zhou J, Wang J, Chen L, Wang L, Hodgkins 
            A, Iyer V, Huang X, Skarnes WC
  TITLE     Efficient genome modification by CRISPR-Cas9 nickase with minimal 
            off-target effects.
  JOURNAL   Nat Methods. 2014 Mar 2. doi: 10.1038/nmeth.2857.
  PUBMED    24584192
REFERENCE   2  (bases 1 to 14884)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 20, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2014 Mar 2. doi: 10.1038/nmeth.2857."
FEATURES             Location/Qualifiers
     source          1..14884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        72..320
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     72..92
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     109..131
                     /label=pRNA-U6Forward
     primer_bind     240..263
                     /label=U6-renyuan
     primer_bind     243..262
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     259..279
                     /label=U6-PROFOR
     primer_bind     complement(287..312)
                     /label=OL533
     primer_bind     293..339
                     /label=OL573
     primer_bind     293..329
                     /label=OL610
     misc_feature    322..341
                     /label=FANCF spacer
     misc_RNA        342..417
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(342..361)
                     /label=OL552
     primer_bind     complement(342..361)
                     /label=OL574
     primer_bind     complement(342..356)
                     /label=OL534
     misc_feature    418..434
                     /label=RT template
     misc_feature    430
                     /label=G to T
     primer_bind     complement(434..449)
                     /label=OL573
     misc_feature    435..447
                     /label=PBS
     misc_feature    455..670
                     /label=H1 promoter
     primer_bind     590..609
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_feature    680..699
                     /label=FANCF nicking
     primer_bind     complement(682..719)
                     /label=OL574
     primer_bind     complement(699..714)
                     /label=OL534
     misc_RNA        700..775
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(700..719)
                     /label=OL552
     misc_feature    827..944
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     857..881
                     /label=U6-shuyuan
     promoter        993..1503
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1422..1440
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             1525..2124
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1525..1544)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2021..2041
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2271..2392
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2274..2293)
                     /label=pCMV5-R
     primer_bind     complement(2282..2301)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2290..2311
                     /label=pFastBac-R
     primer_bind     2336..2355
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2416..2433
                     /label=pEGFP-C3
     primer_bind     complement(2478..2520)
                     /label=PE009
     primer_bind     2501..2548
                     /label=PE007
     enhancer        2505..2884
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(2590..2607)
                     /label=pADTrack-F
     primer_bind     complement(2613..2628)
                     /label=CMV-R
     promoter        2885..3088
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3039..3059
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3049..3066
                     /label=PSilencer4.1-R
     primer_bind     3050..3067
                     /label=pFlag-CMV-F
     primer_bind     3051..3070
                     /label=PIRES2-EGFP.P5
     primer_bind     3062..3081
                     /label=pEGFP-N5
     primer_bind     3070..3089
                     /label=pShuttle-CMV-F
     primer_bind     3130..3149
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3130..3148
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3130..3148
                     /label=T7a
     CDS             3198..3218
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3219..7319
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from 
                     theStreptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(R221K N394K H840A)
                     /note="generates RNA-guided single strand nicks in DNA
                     dueto the H840A mutation in the HNH catalytic domain"
                     /note="/Original_Translation=DKKYSIGLDIGTNSVGWAVITDEYKVPSKK
                     FKVLG 
                     NTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVD 
                     DSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLR 
                     LIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKA 
                     ILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSK 
                     DTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYD 
                     EHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMD 
                     GTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEK 
                     ILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDK 
                     NLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRK 
                     VTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDIL 
                     EDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQ 
                     SGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPA 
                     IKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKE 
                     LGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLK 
                     DDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERG 
                     GLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVS 
                     DFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMI 
                     AKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDF 
                     ATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPT 
                     VAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLII 
                     KLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQ 
                     KQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHL 
                     FTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD"
                     /note="/modified_by=chenp"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRKLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLKREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7320..7421
                     /label=SGGSx2-sv40NLS-SGGSx2
                     /note="/created_by=chenp"
     CDS             7422..9452
                     /label=RT codon optimized Genscript
                     /note="/created_by=jessiedavis"
     primer_bind     complement(9004..9016)
                     /label=OL573
     CDS             9498..9518
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9528..9554
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             9564..9620
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     9564..9587
                     /label=PE001
     primer_bind     complement(9602..9621)
                     /label=PE002
     CDS             9621..11876
                     /label=MLH1 del754-756 genscript codon
                     /note="/created_by=chenp"
                     /note="/modified_by=chenp"
     primer_bind     9662..9704
                     /label=OL530
     primer_bind     9675..9710
                     /label=OL532
     misc_feature    9698
                     /label=A to G
     primer_bind     complement(9981..10019)
                     /label=OL531
     CDS             11885..11902
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(11925..11942)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     primer_bind     complement(11929..11948)
                     /label=PCDNA3.0R-R2
     polyA_signal    11931..12155
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(11975..11992)
                     /label=BGH
     primer_bind     complement(12136..12159)
                     /label=PE008
     primer_bind     12143..12183
                     /label=PE010
     primer_bind     complement(12216..12233)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12387..12975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12467..12486)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(13146..14006)
                     /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     complement(13196..13215)
                     /label=Amp-Ra
     primer_bind     13769..13788
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     13859..13878
                     /label=Amp-F
     promoter        complement(14007..14111)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      14138..14593
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14225..14244)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14435..14456
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     14594..14610
                     /label=puc57-F
     polyA_signal    14724..14772
                     /note="synthetic polyadenylation signal"
     misc_feature    14786..14877
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     14826..14845
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     primer_bind     14827..14845
                     /label=pGL3+
ORIGIN
        1 ggtaccgatt agtgaacgga tctcgacggt atcgatcacg agactagcct cgagcggccg
       61 cccccttcac cgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
      121 ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata
      181 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
      241 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
      301 tgtggaaagg acgaaacacc gggaatccct tctgcagcac cgttttagag ctagaaatag
      361 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcgga
      421 aaagcgatca aggtgctgca gaagggattt ttttgaacgc tgacgtcatc aacccgctcc
      481 aaggaatcgc gggcccagtg tcactaggcg ggaacaccca gcgcgcgtgc gccctggcag
      541 gaagatggct gtgagggaca ggggagtggc gccctgcaat atttgcatgt cgctatgtgt
      601 tctgggaaat caccataaac gtgaaatgtc tttggatttg ggaatcttat aagttctgta
      661 tgaggaccac gaaacaccgg gggtcccagg tgctgacgtg ttttagagct agaaatagca
      721 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
      781 ttaaagaatt ctcgacctcg agacaaatgg cagtattcat ccacaatttt aaaagaaaag
      841 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
      901 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
      961 acagcagaga tccactttgg ccgcggctcg agggggttgg ggttgcgcct tttccaaggc
     1021 agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa acgcagcggc
     1081 gccgaccctg ggactcgcac attcttcacg tccgttcgca gcgtcacccg gatcttcgcc
     1141 gctacccttg tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc
     1201 ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg cacgtctcac tagtaccctc
     1261 gcagacggac agcgccaggg agcaatggca gcgcgccgac cgcgatgggc tgtggccaat
     1321 agcggctgct cagcagggcg cgccgagagc agcggccggg aaggggcggt gcgggaggcg
     1381 gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag
     1441 cctccggagc gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga cctctctccc
     1501 cagggggatc caccggagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc
     1561 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     1621 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     1681 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     1741 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     1801 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     1861 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     1921 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     1981 cgcgccgggg tgcccgcctt cctggaaacc tccgcgcccc gcaacctccc cttctacgag
     2041 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     2101 atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg
     2161 agcgcacgac cccatgcatc ggtaccttta agaccaatga cttacaaggc agctgtagat
     2221 cttagccact ttctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat
     2281 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2341 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     2401 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     2461 gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg tcgagacatt gattattgac
     2521 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     2581 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     2641 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     2701 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     2761 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     2821 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     2881 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     2941 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     3001 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     3061 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagagatc
     3121 cgcggccgct aatacgactc actataggga gagccgccac catgaaacgg acagccgacg
     3181 gaagcgagtt cgagtcacca aagaagaagc ggaaagtcga caagaagtac agcatcggcc
     3241 tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
     3301 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
     3361 gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca
     3421 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
     3481 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
     3541 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
     3601 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
     3661 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
     3721 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
     3781 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
     3841 aggccatcct gtctgccaga ctgagcaaga gcagaaagct ggaaaatctg atcgcccagc
     3901 tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga
     3961 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg
     4021 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
     4081 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
     4141 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc
     4201 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
     4261 agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc
     4321 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac
     4381 tgctcgtgaa gctgaagaga gaggacctgc tgcggaagca gcggaccttc gacaacggca
     4441 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt
     4501 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc
     4561 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga
     4621 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc
     4681 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc
     4741 ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat
     4801 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg
     4861 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
     4921 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
     4981 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
     5041 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
     5101 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
     5161 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
     5221 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
     5281 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
     5341 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
     5401 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
     5461 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
     5521 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
     5581 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
     5641 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
     5701 acatcaaccg gctgtccgac tacgatgtgg acgctatcgt gcctcagagc tttctgaagg
     5761 acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca
     5821 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg
     5881 ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga
     5941 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
     6001 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc
     6061 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
     6121 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
     6181 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg
     6241 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
     6301 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
     6361 agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa
     6421 ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca
     6481 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt
     6541 ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta
     6601 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag
     6661 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca
     6721 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag
     6781 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg
     6841 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc
     6901 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc
     6961 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga
     7021 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca
     7081 aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata
     7141 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca
     7201 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc
     7261 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggtgact
     7321 ccggcggaag ctctggtggc agcaagcgga ccgccgacgg ctctgaattc gagagcccta
     7381 agaagaaaag aaaggtgagc ggaggctcta gcggcggaag caccctgaac attgaagacg
     7441 agtatagact gcatgaaaca agcaaggaac ccgacgtgtc cctgggctcc acctggctgt
     7501 ccgactttcc ccaggcctgg gccgagacag gaggaatggg cctggccgtg cggcaggcac
     7561 ccctgatcat ccctctgaag gccacctcta cacccgtgag catcaagcag taccctatgt
     7621 ctcaggaggc cagactgggc atcaagcctc acatccagag gctgctggac cagggcatcc
     7681 tggtgccatg ccagagcccc tggaacacac cactgctgcc cgtgaagaag ccaggcacca
     7741 atgactatag acccgtgcag gatctgagag aggtgaacaa gagggtggag gatatccacc
     7801 ccaccgtgcc caacccttac aatctgctgt ccggcctgcc cccttctcac cagtggtata
     7861 cagtgctgga cctgaaggat gccttctttt gtctgagact gcaccctacc agccagccac
     7921 tgttcgcctt tgagtggagg gaccctgaga tgggcatctc tggccagctg acctggacac
     7981 gcctgcctca gggcttcaag aatagcccaa cactgtttaa cgaggccctg caccgcgacc
     8041 tggcagattt ccggatccag cacccagatc tgatcctgct gcagtacgtg gacgatctgc
     8101 tgctggccgc caccagcgag ctggattgcc agcagggaac acgcgccctg ctgcagaccc
     8161 tgggaaacct gggatatagg gcatccgcca agaaggccca gatctgtcag aagcaggtga
     8221 agtacctggg ctatctgctg aaggagggcc agagatggct gacagaggcc aggaaggaga
     8281 cagtgatggg ccagccaaca cccaagaccc caagacagct gagggagttc ctgggcaaag
     8341 caggattttg caggctgttc atcccaggat tcgcagagat ggcagcacct ctgtacccac
     8401 tgaccaagcc gggcaccctg tttaattggg gccctgacca gcagaaggcc tatcaggaga
     8461 tcaagcaggc cctgctgaca gcaccagccc tgggcctgcc agacctgacc aagcctttcg
     8521 agctgtttgt ggatgagaag cagggctacg ccaagggcgt gctgacccag aagctgggac
     8581 catggagacg gcccgtggcc tatctgtcca agaagctgga cccagtggca gcaggatggc
     8641 caccatgcct gaggatggtg gcagcaatcg ccgtgctgac aaaggatgcc ggcaagctga
     8701 ccatgggaca gccactggtc atcctggcac cacacgcagt ggaggccctg gtgaagcagc
     8761 ctccagatcg ctggctgtct aacgcccgga tgacacacta ccaggccctg ctgctggaca
     8821 ccgatcgcgt gcagtttggc cctgtggtgg ccctgaatcc agccaccctg ctgcctctgc
     8881 cagaggaggg cctgcagcac aactgtctgg acatcctggc agaggcacac ggaacaaggc
     8941 cagacctgac cgatcagccc ctgcctgacg ccgatcacac atggtatacc gatggaagct
     9001 ccctgctgca ggagggccag aggaaggcag gagcagcagt gaccacagag acagaagtga
     9061 tctgggccaa ggccctgcca gcaggcacat ccgcccagcg ggccgagctg atcgccctga
     9121 cccaggccct gaagatggcc gagggcaaga agctgaacgt gtacacagac tccagatatg
     9181 ccttcgccac cgcacacatc cacggagaga tctacaggcg ccggggctgg ctgacctctg
     9241 agggcaagga gatcaagaac aaggatgaga tcctggccct gctgaaggcc ctgtttctgc
     9301 ccaagcggct gagcatcatc cactgtcctg gacaccagaa gggacactcc gccgaggcaa
     9361 ggggcaatcg gatggccgac caggccgcca gaaaggctgc tattactgaa actcccgaca
     9421 cttccactct gctgattgaa aactcctccc cttctggcgg ctcaaaaaga accgccgacg
     9481 gcagcgaatt cgagtctccc aagaagaaga ggaaagtcgg ctctggccct gccgctaaga
     9541 gagtgaagct ggacggaagc ggagctacta acttcagcct gctgaagcag gctggagacg
     9601 tggaggagaa ccctggacct agcttcgttg ctggagtcat ccggagactg gacgagacag
     9661 tggtgaacag aattgccgcc ggcgaggtga tccagagacc tgccaatgca ataaaggaga
     9721 tgatcgagaa ctgtctggac gccaagtcca caagcattca ggtgatcgtg aaggagggcg
     9781 gactgaagct gatccagatc caagacaacg gcacaggcat cagaaaggaa gatctggaca
     9841 tcgtgtgtga acggttcacc acatctaagc tgcagtcttt tgaggatctg gcctctatca
     9901 gtacctacgg cttcagaggc gaggccctgg ccagcatcag ccacgtggcc catgtgacca
     9961 tcaccaccaa aaccgccgac ggcaaatgcg cttatcgcgc tagctacagc gacggcaagc
    10021 tgaaagcccc gccaaagcct tgcgccggca accagggtac acagataaca gtggaggatc
    10081 tgttctacaa catcgccacc cggagaaagg ccctgaaaaa tcccagcgag gagtacggca
    10141 agatcctgga agtcgtcggc agatactccg tgcacaacgc cggaatcagc tttagcgtaa
    10201 agaagcaggg agaaaccgtg gccgatgtgc gcaccctgcc aaatgccagc accgtggata
    10261 acatcagaag cattttcgga aatgccgtgt ccagagaact gatcgagatc ggctgcgaag
    10321 ataagaccct ggcttttaag atgaacggct acatcagcaa cgccaattac tctgtgaaga
    10381 agtgcatctt tcttctgttc atcaaccaca gactggtgga aagcaccagc ctgcggaaag
    10441 ccatcgagac agtgtacgcc gcctacctgc ctaagaacac ccaccccttc ctgtacctga
    10501 gcctcgagat cagccctcag aacgtggacg tcaatgtgca tcctacaaag cacgaggtgc
    10561 acttcctgca cgaggaaagc atcctggaaa gagtgcagca gcacattgag agcaagctgc
    10621 tgggctctaa cagcagcaga atgtacttca cacagaccct gctgcctggc ctggccggcc
    10681 cctcaggcga aatggttaag tccacaacct ctctgacctc atccagcacc agcggttctt
    10741 ccgataaggt gtacgcccac cagatggtgc ggaccgactc tcgggagcag aagctggacg
    10801 cctttctgca acctctgagc aaacctctga gctctcagcc tcaggccatc gtgaccgagg
    10861 acaagacaga tatctcctcc ggccgtgcca gacagcagga cgaagaaatg ctcgagctgc
    10921 cagctcctgc cgaggtggcc gccaagaacc agagcctgga gggagatacc acaaagggca
    10981 ccagcgaaat gagcgagaag cggggcccta cctccagcaa ccccagaaaa cggcaccggg
    11041 aggacagcga cgtggaaatg gtggaggacg acagcagaaa ggaaatgaca gccgcttgta
    11101 cccctagaag aagaatcatc aacctgacct ccgtgctgag cctgcaggag gagatcaacg
    11161 agcagggcca cgaggtgctg agagagatgc tgcacaatca cagcttcgtg ggctgcgtga
    11221 accctcaatg ggccctggct cagcatcaaa caaagctgta cctgctgaac accaccaagc
    11281 tgagcgaaga gctgttctac cagatcctca tctacgactt cgccaacttc ggcgtgctac
    11341 gcctgagcga gcccgcccct ctgtttgacc tggccatgct ggctctggat agcccagaaa
    11401 gcggctggac agaagaggac ggacctaaag aggggctggc tgaatacatc gtggagttcc
    11461 tgaagaaaaa ggccgagatg ctggccgact acttttctct ggaaatcgac gaggaaggca
    11521 acctgatcgg cctgcctctg ctgatcgata actacgtgcc tcccctggaa ggcctgccca
    11581 tcttcatcct gagactggct acagaggtga actgggacga ggaaaaggaa tgcttcgagt
    11641 ctctgagcaa ggagtgcgcc atgttctata gcatcagaaa acagtacatc tctgaagaga
    11701 gcactctgtc tggccagcag agtgaagtgc ccggaagcat ccccaacagc tggaagtgga
    11761 ccgtggaaca catcgtgtac aaggccctgc ggagccacat tctccctcct aagcacttca
    11821 ccgaggacgg caacatcctg cagctggcca acctgcccga cctttataag gttttctaac
    11881 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
    11941 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
    12001 cactcccact gtcctttcct aataaaatga gaaaattgca tcgcattgtc tgagtaggtg
    12061 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
    12121 tagcaggcat gctggggatg cggtgggctc tatggatgac tgtcttcttt atcatgcaac
    12181 tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
    12241 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
    12301 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
    12361 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
    12421 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
    12481 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
    12541 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
    12601 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
    12661 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
    12721 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
    12781 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
    12841 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
    12901 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
    12961 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
    13021 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
    13081 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
    13141 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
    13201 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
    13261 ggccccagtg ctgcaatgat accgcgggac ccacgctcac cggctccaga tttatcagca
    13321 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
    13381 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
    13441 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
    13501 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
    13561 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
    13621 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
    13681 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
    13741 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
    13801 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
    13861 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
    13921 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
    13981 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
    14041 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
    14101 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta
    14161 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
    14221 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
    14281 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
    14341 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
    14401 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
    14461 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
    14521 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
    14581 acgcttacaa tttgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
    14641 cgggcctctt cgctattacg ccagcccaag ctaccatgat aagtaagtaa tattaaggta
    14701 cgggaggtac ttggagcggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg
    14761 gttttttgtg tgaatcgata gtactaacat acgctctcca tcaaaacaaa acgaaacaaa
    14821 acaaactagc aaaataggct gtccccagtg caagtgcagg tgccagaaca tttctctatc
    14881 gata
//