pCMV-PE5max-EMX1-U6-PegRNA-H1-Nicking sgRNA

LOCUS       pCMV-PE5max-EMX1-U6-PegRNA-H1-Nicking sgRNA              14888 bp DNA     circular SYN 08-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14888)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 21, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..14888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1..25)
                     /label=OL1047
     primer_bind     complement(67..84)
                     /label=pADTrack-F
     primer_bind     complement(90..105)
                     /label=CMV-R
     promoter        362..565
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     516..536
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     526..543
                     /label=PSilencer4.1-R
     primer_bind     527..544
                     /label=pFlag-CMV-F
     primer_bind     528..547
                     /label=PIRES2-EGFP.P5
     primer_bind     539..558
                     /label=pEGFP-N5
     primer_bind     547..566
                     /label=pShuttle-CMV-F
     primer_bind     607..626
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        607..625
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     607..625
                     /label=T7a
     CDS             675..695
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             696..4796
                     /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"
     primer_bind     1748..1767
                     /label=OL2775
     CDS             4797..4898
                     /label=SGGSx2-sv40NLS-SGGSx2
                     /note="/created_by=chenp"
     CDS             4899..6929
                     /label=RT codon optimized Genscript
                     /note="/created_by=jessiedavis"
     primer_bind     complement(5579..5588)
                     /label=CMV-R
     primer_bind     complement(6481..6493)
                     /label=OL573
     primer_bind     6790..6811
                     /label=test
     primer_bind     6863..6885
                     /label=OL447
     CDS             6975..6995
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7005..7031
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7041..7097
                     /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     7041..7064
                     /label=PE001
     primer_bind     complement(7079..7098)
                     /label=PE002
     misc_feature    7098..9356
                     /label=MLH1 del754-756 genscript codon
     CDS             7098..9353
                     /label=MLH1 del754-756 genscript codon
                     /note="/created_by=chenp"
                     /note="/modified_by=chenp"
     primer_bind     7139..7181
                     /label=OL530
     primer_bind     7152..7187
                     /label=OL532
     primer_bind     7160..7190
                     /label=OL485
     primer_bind     complement(7160..7190)
                     /label=OL486
     misc_feature    7175
                     /label=A to G
     misc_feature    7175
                     /label=same sense mutation
     primer_bind     complement(7458..7496)
                     /label=OL531
     primer_bind     9332..9356
                     /label=OL1042
     CDS             9362..9379
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(9402..9419)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     primer_bind     complement(9406..9425)
                     /label=PCDNA3.0R-R2
     polyA_signal    9408..9632
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(9452..9469)
                     /label=BGH
     primer_bind     complement(9613..9636)
                     /label=PE008
     primer_bind     complement(9634..9657)
                     /label=OL1043
     primer_bind     9635..9657
                     /label=OL1044
     rep_origin      complement(9864..10452)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(10623..11483)
                     /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(10673..10692)
                     /label=Amp-R
     primer_bind     11336..11355
                     /label=Amp-F
     promoter        complement(11484..11588)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      11615..12070
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     12071..12087
                     /label=puc57-F
     polyA_signal    12201..12249
                     /note="synthetic polyadenylation signal"
     misc_feature    12263..12354
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     12303..12322
                     /label=RV3
     primer_bind     12304..12322
                     /label=pGL3+
     primer_bind     12388..12403
                     /label=OL2782
     primer_bind     complement(12392..12415)
                     /label=OL459
     promoter        12433..12681
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     12433..12453
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     12604..12623
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     12654..12674
                     /label=OL573
     primer_bind     12679..12704
                     /label=OL2774
     misc_feature    12682..12701
                     /label=EMX1 spacer
     misc_RNA        12702..12777
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    12778..12799
                     /label=RTT
     primer_bind     complement(12792..12830)
                     /label=OL2775
     misc_feature    12800..12812
                     /label=PBS
     primer_bind     12823..12839
                     /label=OL2772
     misc_feature    12825..13035
                     /label=H1 promoter
     primer_bind     12955..12974
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(13027..13068)
                     /label=OL2776
     primer_bind     complement(13027..13044)
                     /label=OL2773
     misc_feature    13045..13064
                     /label=EMX1 nickingRNA
     misc_RNA        13065..13140
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     13141..13164
                     /label=OL2780
     primer_bind     complement(13147..13166)
                     /label=OL2783
     misc_feature    13192..13309
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        13358..13868
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     13787..13805
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             13890..14489
                     /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(13890..13909)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     14386..14406
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(14588..14606)
                     /label=OL2781
     polyA_signal    14636..14757
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(14639..14658)
                     /label=pCMV5-R
     primer_bind     complement(14647..14666)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     14655..14676
                     /label=pFastBac-R
     primer_bind     14701..14720
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     14781..14798
                     /label=pEGFP-C3
     primer_bind     complement(14843..14885)
                     /label=PE009
     primer_bind     join(14866..14888,1..25)
                     /label=PE007
     enhancer        join(14870..14888,1..361)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
       61 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      121 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      181 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      241 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      301 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      361 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      421 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      481 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      541 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agagatccgc
      601 ggccgctaat acgactcact atagggagag ccgccaccat gaaacggaca gccgacggaa
      661 gcgagttcga gtcaccaaag aagaagcgga aagtcgacaa gaagtacagc atcggcctgg
      721 acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca
      781 agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag
      841 ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa
      901 gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga
      961 tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg
     1021 ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg
     1081 agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg
     1141 acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga
     1201 tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc
     1261 agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg
     1321 ccatcctgtc tgccagactg agcaagagca gaaagctgga aaatctgatc gcccagctgc
     1381 ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc
     1441 ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca
     1501 cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt
     1561 ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca
     1621 ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc
     1681 aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga
     1741 ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg
     1801 aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc
     1861 tcgtgaagct gaagagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca
     1921 tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt
     1981 acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct
     2041 actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg
     2101 aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga
     2161 gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca
     2221 agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg
     2281 tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg
     2341 acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca
     2401 agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct
     2461 ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg
     2521 aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag
     2581 agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc
     2641 agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca
     2701 tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca
     2761 acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga
     2821 aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca
     2881 gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag
     2941 tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca
     3001 cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag
     3061 agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga
     3121 agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca
     3181 tcaaccggct gtccgactac gatgtggacg ctatcgtgcc tcagagcttt ctgaaggacg
     3241 actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg
     3301 tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca
     3361 agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg
     3421 aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc
     3481 acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga
     3541 tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt
     3601 tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga
     3661 acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt
     3721 acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg
     3781 gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga
     3841 ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg
     3901 gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc
     3961 cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta
     4021 tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga
     4081 agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg
     4141 aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg
     4201 aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag
     4261 tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc
     4321 ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct
     4381 ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg
     4441 aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca
     4501 tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag
     4561 tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca
     4621 tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca
     4681 ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc
     4741 agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga ggtgactccg
     4801 gcggaagctc tggtggcagc aagcggaccg ccgacggctc tgaattcgag agccctaaga
     4861 agaaaagaaa ggtgagcgga ggctctagcg gcggaagcac cctgaacatt gaagacgagt
     4921 atagactgca tgaaacaagc aaggaacccg acgtgtccct gggctccacc tggctgtccg
     4981 actttcccca ggcctgggcc gagacaggag gaatgggcct ggccgtgcgg caggcacccc
     5041 tgatcatccc tctgaaggcc acctctacac ccgtgagcat caagcagtac cctatgtctc
     5101 aggaggccag actgggcatc aagcctcaca tccagaggct gctggaccag ggcatcctgg
     5161 tgccatgcca gagcccctgg aacacaccac tgctgcccgt gaagaagcca ggcaccaatg
     5221 actatagacc cgtgcaggat ctgagagagg tgaacaagag ggtggaggat atccacccca
     5281 ccgtgcccaa cccttacaat ctgctgtccg gcctgccccc ttctcaccag tggtatacag
     5341 tgctggacct gaaggatgcc ttcttttgtc tgagactgca ccctaccagc cagccactgt
     5401 tcgcctttga gtggagggac cctgagatgg gcatctctgg ccagctgacc tggacacgcc
     5461 tgcctcaggg cttcaagaat agcccaacac tgtttaacga ggccctgcac cgcgacctgg
     5521 cagatttccg gatccagcac ccagatctga tcctgctgca gtacgtggac gatctgctgc
     5581 tggccgccac cagcgagctg gattgccagc agggaacacg cgccctgctg cagaccctgg
     5641 gaaacctggg atatagggca tccgccaaga aggcccagat ctgtcagaag caggtgaagt
     5701 acctgggcta tctgctgaag gagggccaga gatggctgac agaggccagg aaggagacag
     5761 tgatgggcca gccaacaccc aagaccccaa gacagctgag ggagttcctg ggcaaagcag
     5821 gattttgcag gctgttcatc ccaggattcg cagagatggc agcacctctg tacccactga
     5881 ccaagccggg caccctgttt aattggggcc ctgaccagca gaaggcctat caggagatca
     5941 agcaggccct gctgacagca ccagccctgg gcctgccaga cctgaccaag cctttcgagc
     6001 tgtttgtgga tgagaagcag ggctacgcca agggcgtgct gacccagaag ctgggaccat
     6061 ggagacggcc cgtggcctat ctgtccaaga agctggaccc agtggcagca ggatggccac
     6121 catgcctgag gatggtggca gcaatcgccg tgctgacaaa ggatgccggc aagctgacca
     6181 tgggacagcc actggtcatc ctggcaccac acgcagtgga ggccctggtg aagcagcctc
     6241 cagatcgctg gctgtctaac gcccggatga cacactacca ggccctgctg ctggacaccg
     6301 atcgcgtgca gtttggccct gtggtggccc tgaatccagc caccctgctg cctctgccag
     6361 aggagggcct gcagcacaac tgtctggaca tcctggcaga ggcacacgga acaaggccag
     6421 acctgaccga tcagcccctg cctgacgccg atcacacatg gtataccgat ggaagctccc
     6481 tgctgcagga gggccagagg aaggcaggag cagcagtgac cacagagaca gaagtgatct
     6541 gggccaaggc cctgccagca ggcacatccg cccagcgggc cgagctgatc gccctgaccc
     6601 aggccctgaa gatggccgag ggcaagaagc tgaacgtgta cacagactcc agatatgcct
     6661 tcgccaccgc acacatccac ggagagatct acaggcgccg gggctggctg acctctgagg
     6721 gcaaggagat caagaacaag gatgagatcc tggccctgct gaaggccctg tttctgccca
     6781 agcggctgag catcatccac tgtcctggac accagaaggg acactccgcc gaggcaaggg
     6841 gcaatcggat ggccgaccag gccgccagaa aggctgctat tactgaaact cccgacactt
     6901 ccactctgct gattgaaaac tcctcccctt ctggcggctc aaaaagaacc gccgacggca
     6961 gcgaattcga gtctcccaag aagaagagga aagtcggctc tggccctgcc gctaagagag
     7021 tgaagctgga cggaagcgga gctactaact tcagcctgct gaagcaggct ggagacgtgg
     7081 aggagaaccc tggacctagc ttcgttgctg gagtcatccg gagactggac gagacagtgg
     7141 tgaacagaat tgccgccggc gaggtgatcc agagacctgc caatgcaata aaggagatga
     7201 tcgagaactg tctggacgcc aagtccacaa gcattcaggt gatcgtgaag gagggcggac
     7261 tgaagctgat ccagatccaa gacaacggca caggcatcag aaaggaagat ctggacatcg
     7321 tgtgtgaacg gttcaccaca tctaagctgc agtcttttga ggatctggcc tctatcagta
     7381 cctacggctt cagaggcgag gccctggcca gcatcagcca cgtggcccat gtgaccatca
     7441 ccaccaaaac cgccgacggc aaatgcgctt atcgcgctag ctacagcgac ggcaagctga
     7501 aagccccgcc aaagccttgc gccggcaacc agggtacaca gataacagtg gaggatctgt
     7561 tctacaacat cgccacccgg agaaaggccc tgaaaaatcc cagcgaggag tacggcaaga
     7621 tcctggaagt cgtcggcaga tactccgtgc acaacgccgg aatcagcttt agcgtaaaga
     7681 agcagggaga aaccgtggcc gatgtgcgca ccctgccaaa tgccagcacc gtggataaca
     7741 tcagaagcat tttcggaaat gccgtgtcca gagaactgat cgagatcggc tgcgaagata
     7801 agaccctggc ttttaagatg aacggctaca tcagcaacgc caattactct gtgaagaagt
     7861 gcatctttct tctgttcatc aaccacagac tggtggaaag caccagcctg cggaaagcca
     7921 tcgagacagt gtacgccgcc tacctgccta agaacaccca ccccttcctg tacctgagcc
     7981 tcgagatcag ccctcagaac gtggacgtca atgtgcatcc tacaaagcac gaggtgcact
     8041 tcctgcacga ggaaagcatc ctggaaagag tgcagcagca cattgagagc aagctgctgg
     8101 gctctaacag cagcagaatg tacttcacac agaccctgct gcctggcctg gccggcccct
     8161 caggcgaaat ggttaagtcc acaacctctc tgacctcatc cagcaccagc ggttcttccg
     8221 ataaggtgta cgcccaccag atggtgcgga ccgactctcg ggagcagaag ctggacgcct
     8281 ttctgcaacc tctgagcaaa cctctgagct ctcagcctca ggccatcgtg accgaggaca
     8341 agacagatat ctcctccggc cgtgccagac agcaggacga agaaatgctc gagctgccag
     8401 ctcctgccga ggtggccgcc aagaaccaga gcctggaggg agataccaca aagggcacca
     8461 gcgaaatgag cgagaagcgg ggccctacct ccagcaaccc cagaaaacgg caccgggagg
     8521 acagcgacgt ggaaatggtg gaggacgaca gcagaaagga aatgacagcc gcttgtaccc
     8581 ctagaagaag aatcatcaac ctgacctccg tgctgagcct gcaggaggag atcaacgagc
     8641 agggccacga ggtgctgaga gagatgctgc acaatcacag cttcgtgggc tgcgtgaacc
     8701 ctcaatgggc cctggctcag catcaaacaa agctgtacct gctgaacacc accaagctga
     8761 gcgaagagct gttctaccag atcctcatct acgacttcgc caacttcggc gtgctacgcc
     8821 tgagcgagcc cgcccctctg tttgacctgg ccatgctggc tctggatagc ccagaaagcg
     8881 gctggacaga agaggacgga cctaaagagg ggctggctga atacatcgtg gagttcctga
     8941 agaaaaaggc cgagatgctg gccgactact tttctctgga aatcgacgag gaaggcaacc
     9001 tgatcggcct gcctctgctg atcgataact acgtgcctcc cctggaaggc ctgcccatct
     9061 tcatcctgag actggctaca gaggtgaact gggacgagga aaaggaatgc ttcgagtctc
     9121 tgagcaagga gtgcgccatg ttctatagca tcagaaaaca gtacatctct gaagagagca
     9181 ctctgtctgg ccagcagagt gaagtgcccg gaagcatccc caacagctgg aagtggaccg
     9241 tggaacacat cgtgtacaag gccctgcgga gccacattct ccctcctaag cacttcaccg
     9301 aggacggcaa catcctgcag ctggccaacc tgcccgacct ttataaggtt ttctaaccgg
     9361 tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctag
     9421 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     9481 tcccactgtc ctttcctaat aaaatgagaa aattgcatcg cattgtctga gtaggtgtca
     9541 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     9601 caggcatgct ggggatgcgg tgggctctat ggatgactgt cttctttatc atgcaactcg
     9661 taggacaggt gccggcagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     9721 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     9781 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     9841 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     9901 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     9961 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
    10021 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
    10081 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
    10141 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
    10201 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
    10261 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
    10321 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
    10381 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
    10441 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
    10501 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
    10561 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
    10621 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
    10681 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
    10741 cccagtgctg caatgatacc gcgggaccca cgctcaccgg ctccagattt atcagcaata
    10801 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
    10861 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
    10921 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
    10981 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
    11041 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
    11101 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
    11161 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
    11221 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
    11281 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
    11341 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
    11401 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
    11461 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
    11521 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
    11581 gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc
    11641 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
    11701 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
    11761 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
    11821 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
    11881 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
    11941 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
    12001 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
    12061 cttacaattt gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
    12121 gcctcttcgc tattacgcca gcccaagcta ccatgataag taagtaatat taaggtacgg
    12181 gaggtacttg gagcggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
    12241 ttttgtgtga atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca
    12301 aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat
    12361 aggtaccgat tagtgaacgg atctcgacgg tatcgatcac gagactagcc tcgagcggcc
    12421 gcccccttca ccgagggcct atttcccatg attccttcat atttgcatat acgatacaag
    12481 gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat
    12541 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
    12601 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
    12661 ttgtggaaag gacgaaacac cgagtccgag cagaagaaga agttttagag ctagaaatag
    12721 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcgtt
    12781 gatgtgatgg gagccgttct tcttctgctc ggtttttttg aacgctgacg tcatcaaccc
    12841 gctccaagga atcgcgggcc cagtgtcact aggcgggaac acccagcgcg cgtgcgccct
    12901 ggcaggaaga tggctgtgag ggacagggga gtggcgccct gcaatatttg catgtcgcta
    12961 tgtgttctgg gaaatcacca taaacgtgaa atgtctttgg atttgggaat cttataagtt
    13021 ctgtatgagg accacgaaac accgctcccc attggcctgc ttcggtttta gagctagaaa
    13081 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
    13141 tttttttaaa gaattctcga cctcgagaca aatggcagta ttcatccaca attttaaaag
    13201 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
    13261 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
    13321 cagggacagc agagatccac tttggccgcg gctcgagggg gttggggttg cgccttttcc
    13381 aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc gggaaacgca
    13441 gcggcgccga ccctgggact cgcacattct tcacgtccgt tcgcagcgtc acccggatct
    13501 tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta agtcgggaag
    13561 gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt ctcactagta
    13621 ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga tgggctgtgg
    13681 ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg gcggtgcggg
    13741 aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt tccgcattct
    13801 gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc accgacctct
    13861 ctccccaggg ggatccaccg gagcttacca tgaccgagta caagcccacg gtgcgcctcg
    13921 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
    13981 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
    14041 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
    14101 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
    14161 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
    14221 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
    14281 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
    14341 ccgagcgcgc cggggtgccc gccttcctgg aaacctccgc gccccgcaac ctccccttct
    14401 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
    14461 ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg
    14521 aaaggagcgc acgaccccat gcatcggtac ctttaagacc aatgacttac aaggcagctg
    14581 tagatcttag ccactttcta gagtcggggc ggccggccgc ttcgagcaga catgataaga
    14641 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
    14701 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
    14761 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
    14821 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccgtcgag acattgatta
    14881 ttgactag
//