LOCUS       sequence-298673-       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  JOURNAL   Unpublished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            "Unpublished"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 g1 + Cas9
FEATURES             Location/Qualifiers
     source          1..11317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    8..24
                     /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     13..35
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     32..48
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     32..48
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     67..87
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        69..87
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        115..341
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             342..522
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    569..694
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1187..1420
                     /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(1599..1847)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(1657..1676)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1827..1847)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    1854..1971
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2040..2251
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2271..2280
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2280..2303
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2310..2330
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2355..6455
                     /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             2355..6455
                     /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             6456..6503
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6519..6575
                     /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             6576..7277
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    7284..7872
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7337..7357)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7755..7766)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7944..8177
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8255..8376
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8292..8311)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8346..8365
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8416..8551
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8571..8590)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8572..8590)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8600..8617)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8600..8616)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8609..8631)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8758..9213
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8845..8864)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9055..9076
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9239..9343
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9344..9412,9413..10204)
                     /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(9562..9581)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10375..10963
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10864..10883
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11117..11134
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11251..11272
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11287..11292,11293..11310,11311..11317)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
       61 aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt
      121 cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa catgccttac
      181 aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac gatcgtgcct
      241 tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa ttgccgcatt
      301 gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc tggttagacc
      361 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
      421 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
      481 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
      541 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
      601 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
      661 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
      721 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
      781 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
      841 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
      901 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
      961 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
     1021 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     1081 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     1141 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     1201 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     1261 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     1321 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     1381 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     1441 gaaaactcgc catttgtctc aagatctaga attcaaaaaa agcaccgact cggtgccact
     1501 ttgctgtttc cagcaaagtt gataacggac tagccttatt taaacttgct atgctgtttc
     1561 cagcatagct cttaaacgaa gtcgcccttc cccaggtcgg tgtttcgtcc tttccacaag
     1621 atatataaag ccaagaaatc gaaatacttt caagttacgg taagcatatg atagtccatt
     1681 ttaaaacata attttaaaac tgcaaactac ccaagaaatt attactttct acgtcacgta
     1741 ttttgtacta atatctttgt gtttacagtc aaattaattc taattatctc tctaacagcc
     1801 ttgtatcgta tatgcaaata tgaaggaatc atgggaaata ggccctccca caattttaaa
     1861 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
     1921 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat
     1981 tacagggaca gcagagatcc actttggcgc cggctcgagt ggctccggtg cccgtcagtg
     2041 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
     2101 cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
     2161 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
     2221 ttttcgcaac gggtttgccg ccagaacaca ggtgtcgtga cgcgggatcc gccaccatgg
     2281 attacaaaga cgatgacgat aagatggccc caaagaagaa gcggaaggtc ggtatccacg
     2341 gagtcccagc agccgacaag aagtacagca tcggcctgga catcggcacc aactctgtgg
     2401 gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca
     2461 acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg
     2521 aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga
     2581 accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct
     2641 tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc
     2701 ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc
     2761 acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg
     2821 ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg
     2881 acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg
     2941 aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga
     3001 gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc
     3061 tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg
     3121 acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca
     3181 acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt
     3241 ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc
     3301 tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag
     3361 ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga
     3421 acggctacgc cggctacatt gacggcggag ccagccagga agagttctac aagttcatca
     3481 agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg
     3541 acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg
     3601 gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc
     3661 gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca
     3721 ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga
     3781 acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca
     3841 acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt
     3901 acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc
     3961 ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc
     4021 ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact
     4081 ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc
     4141 tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg
     4201 aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga
     4261 aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca
     4321 ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca
     4381 agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga
     4441 tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg
     4501 gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca
     4561 tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg
     4621 agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca
     4681 gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga
     4741 aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc
     4801 agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg
     4861 atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc
     4921 tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga
     4981 agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt
     5041 tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca
     5101 tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact
     5161 cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca
     5221 ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg
     5281 agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc
     5341 tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg
     5401 acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact
     5461 tcttctacag caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga
     5521 tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg
     5581 gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa
     5641 agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg
     5701 ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc
     5761 ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac
     5821 tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga
     5881 atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca
     5941 agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg
     6001 ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt
     6061 acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc
     6121 tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct
     6181 ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc
     6241 accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca
     6301 atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca
     6361 ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg
     6421 agacacggat cgacctgtct cagctgggag gcgacaagcg tcctgctgct actaagaaag
     6481 ctggtcaagc taagaaaaag aaagctagcg gcagcggcgc caccaacttc agcctgctga
     6541 agcaggccgg cgacgtggag gagaaccccg gccccatgag cgagctgatt aaggagaaca
     6601 tgcacatgaa gctgtacatg gagggcaccg tggacaacca tcacttcaag tgcacatccg
     6661 agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg gtcgagggcg
     6721 gccctctccc cttcgccttc gacatcctgg ctactagctt cctctacggc agcaagacct
     6781 tcatcaacca cacccagggc atccccgact tcttcaagca gtccttccct gagggcttca
     6841 catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca
     6901 gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcacatcca
     6961 acggccctgt gatgcagaag aaaacactcg gctgggaggc cttcaccgaa acgctgtacc
     7021 ccgctgacgg cggcctggaa ggcagaaacg acatggccct gaagctcgtg ggcgggagcc
     7081 atctgatcgc aaacgccaag accacatata gatccaagaa acccgctaag aacctcaaga
     7141 tgcctggcgt ctactatgtg gactacagac tggaaagaat caaggaggcc aacaacgaga
     7201 cctacgtcga gcagcacgag gtggcagtgg ccagatactg cgacctccct agcaaactgg
     7261 ggcacaagct taattgagtc gacaatcaac ctctggatta caaaatttgt gaaagattga
     7321 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     7381 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     7441 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     7501 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     7561 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     7621 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     7681 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     7741 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     7801 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     7861 ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag atcttagcca
     7921 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct
     7981 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     8041 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     8101 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     8161 gtggaaaatc tctagcagta cgtatagtag ttcatgtcat cttattattc agtatttata
     8221 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
     8281 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     8341 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
     8401 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     8461 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     8521 cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg
     8581 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     8641 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     8701 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
     8761 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     8821 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     8881 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     8941 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     9001 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     9061 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     9121 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     9181 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     9241 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     9301 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     9361 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     9421 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     9481 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     9541 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     9601 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     9661 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     9721 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     9781 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     9841 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     9901 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     9961 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
    10021 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
    10081 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
    10141 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
    10201 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
    10261 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
    10321 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
    10381 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
    10441 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
    10501 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
    10561 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
    10621 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
    10681 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
    10741 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
    10801 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
    10861 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
    10921 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
    10981 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
    11041 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
    11101 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
    11161 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
    11221 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
    11281 ccaggcttta cactttatgc ttccggctcg tatgttg
//
