LOCUS       sequence-298677-       11317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A guide RNA targeting ACE2 in a lentiviral plasmid co-expressing 
            Cas9 and TagBFP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11317)
  TITLE     CRISPR plasmids to knock out SARS-CoV-2 host genes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11317)
  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: 
            "Unpublished"
COMMENT     Sequence Label: ACE2 g4 + Cas9
FEATURES             Location/Qualifiers
     source          1..11317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     promoter        complement(1485..1733)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(1543..1562)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1713..1733)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    1740..1857
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1926..2137
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2157..2166
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2166..2189
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2196..2216
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2241..6341
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             2241..6341
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6342..6389
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6405..6461
                     /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"
     CDS             6462..7163
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    7170..7758
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7223..7243)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7641..7652)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7830..8063
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8141..8262
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8178..8197)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8232..8251
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8302..8437
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8457..8476)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8458..8476)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8486..8503)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8486..8502)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8495..8517)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8644..9099
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8731..8750)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8941..8962
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9125..9229
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9230..9298,9299..10090)
                     /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(9448..9467)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10261..10849
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10750..10769
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11003..11020
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11137..11158
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11173..11178,11179..11196,11197..11203)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11211..11227
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     11216..11238
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11235..11251
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11235..11251
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11270..11290
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11272..11290
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcgccatt tgtctcaaga tctagaattc aaaaaaagca ccgactcggt
     1381 gccactttgc tgtttccagc aaagttgata acggactagc cttatttaaa cttgctatgc
     1441 tgtttccagc atagctctta aaccacttgc ccaaatgtat ccacggtgtt tcgtcctttc
     1501 cacaagatat ataaagccaa gaaatcgaaa tactttcaag ttacggtaag catatgatag
     1561 tccattttaa aacataattt taaaactgca aactacccaa gaaattatta ctttctacgt
     1621 cacgtatttt gtactaatat ctttgtgttt acagtcaaat taattctaat tatctctcta
     1681 acagccttgt atcgtatatg caaatatgaa ggaatcatgg gaaataggcc ctcccacaat
     1741 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
     1801 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg
     1861 gtttattaca gggacagcag agatccactt tggcgccggc tcgagtggct ccggtgcccg
     1921 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     1981 ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     2041 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     2101 cgttcttttt cgcaacgggt ttgccgccag aacacaggtg tcgtgacgcg ggatccgcca
     2161 ccatggatta caaagacgat gacgataaga tggccccaaa gaagaagcgg aaggtcggta
     2221 tccacggagt cccagcagcc gacaagaagt acagcatcgg cctggacatc ggcaccaact
     2281 ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc
     2341 tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca
     2401 gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac
     2461 ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg
     2521 acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc
     2581 ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca
     2641 tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct
     2701 atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga
     2761 accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc
     2821 tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca
     2881 gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga
     2941 atggcctgtt cggaaacctg attgccctga gcctgggcct gacccccaac ttcaagagca
     3001 acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc
     3061 tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga
     3121 acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg
     3181 cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc
     3241 tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga
     3301 gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt
     3361 tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca
     3421 gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc
     3481 acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg
     3541 acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc
     3601 tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc
     3661 cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga
     3721 tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt
     3781 acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga
     3841 gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga
     3901 ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct
     3961 tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc
     4021 acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca
     4081 ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac
     4141 ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga
     4201 gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt
     4261 ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc
     4321 agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg
     4381 gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga
     4441 agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca
     4501 agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga
     4561 agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga
     4621 tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact
     4681 acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg
     4741 actacgatgt ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca
     4801 aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg
     4861 tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga
     4921 gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg
     4981 gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc
     5041 tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag
     5101 tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag
     5161 tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa
     5221 ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg
     5281 tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca
     5341 agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg
     5401 gcgagatccg gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg
     5461 ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg
     5521 tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga
     5581 acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg
     5641 acagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca
     5701 agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg
     5761 agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga
     5821 tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg
     5881 cctctgccgg cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact
     5941 tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga
     6001 aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg
     6061 agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca
     6121 acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc
     6181 tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga
     6241 ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc
     6301 tgtacgagac acggatcgac ctgtctcagc tgggaggcga caagcgtcct gctgctacta
     6361 agaaagctgg tcaagctaag aaaaagaaag ctagcggcag cggcgccacc aacttcagcc
     6421 tgctgaagca ggccggcgac gtggaggaga accccggccc catgagcgag ctgattaagg
     6481 agaacatgca catgaagctg tacatggagg gcaccgtgga caaccatcac ttcaagtgca
     6541 catccgaggg cgaaggcaag ccctacgagg gcacccagac catgagaatc aaggtggtcg
     6601 agggcggccc tctccccttc gccttcgaca tcctggctac tagcttcctc tacggcagca
     6661 agaccttcat caaccacacc cagggcatcc ccgacttctt caagcagtcc ttccctgagg
     6721 gcttcacatg ggagagagtc accacatacg aagacggggg cgtgctgacc gctacccagg
     6781 acaccagcct ccaggacggc tgcctcatct acaacgtcaa gatcagaggg gtgaacttca
     6841 catccaacgg ccctgtgatg cagaagaaaa cactcggctg ggaggccttc accgaaacgc
     6901 tgtaccccgc tgacggcggc ctggaaggca gaaacgacat ggccctgaag ctcgtgggcg
     6961 ggagccatct gatcgcaaac gccaagacca catatagatc caagaaaccc gctaagaacc
     7021 tcaagatgcc tggcgtctac tatgtggact acagactgga aagaatcaag gaggccaaca
     7081 acgagaccta cgtcgagcag cacgaggtgg cagtggccag atactgcgac ctccctagca
     7141 aactggggca caagcttaat tgagtcgaca atcaacctct ggattacaaa atttgtgaaa
     7201 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     7261 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     7321 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     7381 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     7441 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     7501 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     7561 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     7621 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     7681 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     7741 tttgggccgc ctccccgcgt cgactttaag accaatgact tacaaggcag ctgtagatct
     7801 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     7861 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     7921 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     7981 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     8041 gtcagtgtgg aaaatctcta gcagtacgta tagtagttca tgtcatctta ttattcagta
     8101 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
     8161 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     8221 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     8281 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     8341 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     8401 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct
     8461 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     8521 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     8581 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     8641 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     8701 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     8761 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     8821 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     8881 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     8941 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     9001 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     9061 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     9121 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     9181 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     9241 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     9301 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     9361 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     9421 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     9481 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     9541 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     9601 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     9661 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     9721 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     9781 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     9841 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     9901 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     9961 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
    10021 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
    10081 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
    10141 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
    10201 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
    10261 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
    10321 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
    10381 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
    10441 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
    10501 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
    10561 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
    10621 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
    10681 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
    10741 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
    10801 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
    10861 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
    10921 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
    10981 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
    11041 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
    11101 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
    11161 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
    11221 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
    11281 tcactaaagg gaacaaaagc tggagctgca agcttaa
//
