LOCUS       sequence-298698-       11317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A guide RNA targeting TMPRSS2 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  JOURNAL   Unpublished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            "Unpublished"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: TMPRSS2 g4 + Cas9
FEATURES             Location/Qualifiers
     source          1..11317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    846..1079
                     /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(1258..1506)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(1316..1335)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1486..1506)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    1513..1630
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1699..1910
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      1930..1939
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1939..1962
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1969..1989
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2014..6114
                     /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             2014..6114
                     /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             6115..6162
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6178..6234
                     /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             6235..6936
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    6943..7531
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6996..7016)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7414..7425)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7603..7836
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7914..8035
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7951..7970)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8005..8024
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8075..8210
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8230..8249)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8231..8249)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8259..8276)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8259..8275)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8268..8290)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8417..8872
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8504..8523)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8714..8735
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8898..9002
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9003..9071,9072..9863)
                     /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(9221..9240)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10034..10622
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10523..10542
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10776..10793
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10910..10931
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10946..10951,10952..10969,10970..10976)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10984..11000
                     /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     10989..11011
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11008..11024
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11008..11024
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11043..11063
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11045..11063
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        11091..11317
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcgcc atttgtctca agatctagaa ttcaaaaaaa
     1141 gcaccgactc ggtgccactt tgctgtttcc agcaaagttg ataacggact agccttattt
     1201 aaacttgcta tgctgtttcc agcatagctc ttaaacctta acaatccatg gcattgcggt
     1261 gtttcgtcct ttccacaaga tatataaagc caagaaatcg aaatactttc aagttacggt
     1321 aagcatatga tagtccattt taaaacataa ttttaaaact gcaaactacc caagaaatta
     1381 ttactttcta cgtcacgtat tttgtactaa tatctttgtg tttacagtca aattaattct
     1441 aattatctct ctaacagcct tgtatcgtat atgcaaatat gaaggaatca tgggaaatag
     1501 gccctcccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
     1561 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
     1621 tcaaaatttt cgggtttatt acagggacag cagagatcca ctttggcgcc ggctcgagtg
     1681 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     1741 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     1801 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     1861 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtgtcgtgac
     1921 gcgggatccg ccaccatgga ttacaaagac gatgacgata agatggcccc aaagaagaag
     1981 cggaaggtcg gtatccacgg agtcccagca gccgacaaga agtacagcat cggcctggac
     2041 atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag
     2101 aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc
     2161 ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga
     2221 agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg
     2281 gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat
     2341 aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag
     2401 aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac
     2461 ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc
     2521 gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag
     2581 acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc
     2641 atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc
     2701 ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc
     2761 aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc
     2821 tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt
     2881 ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc
     2941 gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag
     3001 gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt
     3061 ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa
     3121 gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc
     3181 gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc
     3241 ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac
     3301 ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac
     3361 tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag
     3421 gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc
     3481 ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag
     3541 cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg
     3601 accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac
     3661 ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag
     3721 aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc
     3781 ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag
     3841 gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag
     3901 atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag
     3961 ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc
     4021 cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac
     4081 agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa
     4141 gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc
     4201 cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg
     4261 atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc
     4321 cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag
     4381 ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag
     4441 ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc
     4501 aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac
     4561 tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg
     4621 ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag
     4681 ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa
     4741 ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac
     4801 gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc
     4861 cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc
     4921 cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac
     4981 gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac
     5041 ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc
     5101 aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt
     5161 accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg
     5221 gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc
     5281 caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc
     5341 ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag
     5401 tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa
     5461 aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa
     5521 agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg
     5581 aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg
     5641 aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc
     5701 aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag
     5761 gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc
     5821 gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg
     5881 ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc
     5941 cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc
     6001 atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag
     6061 agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg cgacaagcgt
     6121 cctgctgcta ctaagaaagc tggtcaagct aagaaaaaga aagctagcgg cagcggcgcc
     6181 accaacttca gcctgctgaa gcaggccggc gacgtggagg agaaccccgg ccccatgagc
     6241 gagctgatta aggagaacat gcacatgaag ctgtacatgg agggcaccgt ggacaaccat
     6301 cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga
     6361 atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc tactagcttc
     6421 ctctacggca gcaagacctt catcaaccac acccagggca tccccgactt cttcaagcag
     6481 tccttccctg agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg
     6541 accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga
     6601 ggggtgaact tcacatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc
     6661 ttcaccgaaa cgctgtaccc cgctgacggc ggcctggaag gcagaaacga catggccctg
     6721 aagctcgtgg gcgggagcca tctgatcgca aacgccaaga ccacatatag atccaagaaa
     6781 cccgctaaga acctcaagat gcctggcgtc tactatgtgg actacagact ggaaagaatc
     6841 aaggaggcca acaacgagac ctacgtcgag cagcacgagg tggcagtggc cagatactgc
     6901 gacctcccta gcaaactggg gcacaagctt aattgagtcg acaatcaacc tctggattac
     6961 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     7021 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     7081 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     7141 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     7201 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     7261 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     7321 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     7381 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     7441 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     7501 agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg
     7561 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     7621 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     7681 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     7741 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     7801 tcagaccctt ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc
     7861 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     7921 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     7981 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     8041 tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     8101 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     8161 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta
     8221 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     8281 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     8341 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     8401 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     8461 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     8521 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     8581 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     8641 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     8701 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     8761 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     8821 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     8881 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     8941 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     9001 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     9061 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     9121 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     9181 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     9241 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     9301 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     9361 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     9421 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     9481 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     9541 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     9601 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     9661 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     9721 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     9781 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     9841 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     9901 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     9961 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
    10021 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
    10081 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
    10141 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
    10201 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
    10261 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
    10321 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
    10381 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
    10441 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
    10501 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
    10561 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
    10621 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
    10681 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
    10741 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
    10801 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
    10861 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
    10921 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
    10981 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
    11041 gcgcaattaa ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta
    11101 tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg
    11161 agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt
    11221 aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag
    11281 agatattgta tttaagtgcc tagctcgata cataaac
//
