LOCUS       sequence-365005-       11590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus all in one CRISPR vector targeting human AAVS1 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11590)
  AUTHORS   Tsvetkov P, Coy S, Petrova B, Dreishpoon M, Verma A, Abdusamad M, 
            Rossen J, Joesch-Cohen L, Humeidi R, Spangler RD, Eaton JK, Frenkel 
            E, Kocak M, Corsello SM, Lutsenko S, Kanarek N, Santagata S, Golub 
            TR
  TITLE     Copper induces cell death by targeting lipoylated TCA cycle 
            proteins.
  JOURNAL   Science. 2022 Mar 18;375(6586):1254-1261. doi: 
            10.1126/science.abf0529. Epub 2022 Mar 17.
  PUBMED    35298263
REFERENCE   2  (bases 1 to 11590)
  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"; doi: 
            "10.1126/science.abf0529"; journalName: "Science"; date: 
            "2022-03-18- 18"; volume: "375"; issue: "6586"; pages: "1254-1261"
COMMENT     Sequence Label: plentiCRISPv2_AAVS1
FEATURES             Location/Qualifiers
     source          1..11590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     41..60
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             175..408
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    480..1068
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             586..597
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     995..1015
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1084..1683)
                     /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(1167..1187)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1664..1683
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(1684..1740)
                     /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             complement(1756..1803)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(1804..5904)
                     /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             complement(1804..5904)
                     /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             complement(5929..5949)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(5956..5979)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      5979..5988
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6008..6219)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    6288..6405
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        complement(6453..6528)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(6550..6798)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(6608..6627)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(6778..6798)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(6976..7020)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7205..7438
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    7931..8056
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             8103..8283
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(8284..8510)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(8538..8558)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8538..8556)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8577..8593)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8577..8593)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8590..8612)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8601..8617
                     /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)."
     promoter        complement(join(8625..8631,8632..8649,8650..8655))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8670..8691
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8808..8825)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8979..9567)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9059..9078)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9738..10529,10530..10598))
                     /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     10361..10380
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10599..10703)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10729..11184)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10866..10887)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     11078..11097
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11311..11333
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11325..11342
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11326..11342
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11352..11371
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        11352..11370
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      11391..11526
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    join(11566..11590,1..97)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(join(11577..11590,1..6))
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
       61 tttatttgta accattataa gctgcaataa acaagttcct ctcactctct gatattcatt
      121 tctttgcaag ttataaatac tgaataataa gatgacatga actactatac gtactgctag
      181 agattttcca cactgactaa aagggtctga gggatctcta gttaccagag tcacacaaca
      241 gacgggcaca cactacttga agcactcaag gcaagcttta ttgaggctta agcagtgggt
      301 tccctagtta gccagagagc tcccaggctc agatctggtc taaccagaga gacccagtac
      361 aagcaaaaag cagatcttgt cttcgttggg agtgaattag cccttccagt cccccctttt
      421 cttttaaaaa gtggctaaga tctacagctg ccttgtaagt cattggtctt aaagtcgacg
      481 cggggaggcg gcccaaaggg agatccgact cgtctgaggg cgaaggcgaa gacgcggaag
      541 aggccgcaga gccggcagca ggccgcggga aggaaggtcc gctggattga gggccgaagg
      601 gacgtagcag aaggacgtcc cgcgcagaat ccaggtggca acacaggcga gcagccaagg
      661 aaaggacgat gatttccccg acaacaccac ggaattgtca gtgcccaaca gccgagcccc
      721 tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaatag ggagggggaa
      781 agcgaaagtc ccggaaagga gctgacaggt ggtggcaatg ccccaaccag tgggggttgc
      841 gtcagcaaac acagtgcaca ccacgccacg ttgcctgaca acgggccaca actcctcata
      901 aagagacagc aaccaggatt tatacaagga ggagaaaatg aaagccatac gggaagcaat
      961 agcatgatac aaaggcatta aagcagcgta tccacatagc gtaaaaggag caacatagtt
     1021 aagaatacca gtcaatcttt cacaaatttt gtaatccaga ggttgattgt cgacttaacg
     1081 cgttcaggca ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg gcacctcgac
     1141 gtcggcggtg acggtgaagc cgagccgctc gtagaagggg aggttgcggg gcgcggaggt
     1201 ctccaggaag gcgggcaccc cggcgcgctc ggccgcctcc actccgggga gcacgacggc
     1261 gctgcccaga cccttgccct ggtggtcggg cgagactccg acggtggcca ggaaccacgc
     1321 gggctccttg ggccggtgcg gcgccaggag gccttccatc tgttgctgcg cggccagccg
     1381 ggaaccgctc aactcggcca tgcgcgggcc gatctcggcg aacaccgccc ccgcttcgac
     1441 gctctccggc gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc acaccttgcc
     1501 gatgtcgagc ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc gctcgatgtg
     1561 gcggtccgga tcgacggtgt ggcgcgtggc ggggtagtcg gcgaacgcgg cggcgagggt
     1621 gcgtacggcc ctggggacgt cgtcgcgggt ggcgaggcgc accgtgggct tgtactcggt
     1681 catggggccg gggttctcct ccacgtcgcc ggcctgcttc agcaggctga agttggtggc
     1741 gccgctgccg ctagctttct ttttcttagc ttgaccagct ttcttagtag cagcaggacg
     1801 cttgtcgcct cccagctgag acaggtcgat ccgtgtctcg tacaggccgg tgatgctctg
     1861 gtggatcagg gtggcgtcca gcacctcttt ggtgctggtg tacctcttcc ggtcgatggt
     1921 ggtgtcaaag tacttgaagg cggcaggggc tcccagattg gtcagggtaa acaggtggat
     1981 gatattctcg gcctgctctc tgatgggctt atcccggtgc ttgttgtagg cggacagcac
     2041 tttgtccaga ttagcgtcgg ccaggatcac tctcttggag aactcgctga tctgctcgat
     2101 gatctcgtcc aggtagtgct tgtgctgttc cacaaacagc tgtttctgct cattatcctc
     2161 gggggagccc ttcagcttct catagtggct ggccaggtac aggaagttca catatttgga
     2221 gggcagggcc agttcgtttc ccttctgcag ttcgccggca gaggccagca ttctcttccg
     2281 gccgttttcc agctcgaaca gggagtactt aggcagcttg atgatcaggt cctttttcac
     2341 ttctttgtag cccttggctt ccagaaagtc gatgggattc ttctcgaagc tgcttctttc
     2401 catgatggtg atccccagca gctctttcac actcttcagt ttcttggact tgcccttttc
     2461 cactttggcc accaccagca cagaataggc cacggtgggg ctgtcgaagc cgccgtactt
     2521 cttagggtcc cagtccttct ttctggcgat cagcttatcg ctgttcctct tgggcaggat
     2581 agactctttg ctgaagccgc ctgtctgcac ctcggtcttt ttcacgatat tcacttgggg
     2641 catgctcagc actttccgca cggtggcaaa atcccggccc ttatcccaca cgatctcccc
     2701 ggtttcgccg tttgtctcga tcagaggccg cttccggatc tcgccgttgg ccagggtaat
     2761 ctcggtcttg aaaaagttca tgatgttgct gtagaagaag tacttggcgg tagccttgcc
     2821 gatttcctgc tcgctcttgg cgatcatctt ccgcacgtcg tacaccttgt agtcgccgta
     2881 cacgaactcg ctttccagct tagggtactt tttgatcagg gcggttccca cgacggcgtt
     2941 caggtaggcg tcgtgggcgt ggtggtagtt gttgatctcg cgcactttgt aaaactggaa
     3001 atccttccgg aaatcggaca ccagcttgga cttcagggtg atcactttca cttcccggat
     3061 cagcttgtca ttctcgtcgt acttagtgtt catccgggag tccaggatct gtgccacgtg
     3121 ctttgtgatc tgccgggttt ccaccagctg tctcttgatg aagccggcct tatccagttc
     3181 gctcaggccg cctctctcgg ccttggtcag attgtcgaac tttctctggg taatcagctt
     3241 ggcgttcagc agctgccgcc agtagttctt catcttcttc acgacctctt cggagggcac
     3301 gttgtcgctc ttgccccggt tcttgtcgct tctggtcagc accttgttgt cgatggagtc
     3361 gtccttcaga aagctctgag gcacgatatg gtccacatcg tagtcggaca gccggttgat
     3421 gtccagttcc tggtccacgt acatatcccg cccattctgc aggtagtaca ggtacagctt
     3481 ctcgttctgc agctgggtgt tttccacggg gtgttctttc aggatctggc tgcccagctc
     3541 tttgatgccc tcttcgatcc gcttcattct ctcgcggctg ttcttctgtc ccttctgggt
     3601 ggtctggttc tctctggcca tttcgatcac gatgttctcg ggcttgtgcc ggcccatcac
     3661 tttcacgagc tcgtccacca ccttcactgt ctgcaggatg cccttcttaa tggcggggct
     3721 gccggccaga ttggcaatgt gctcgtgcag gctatcgccc tggccggaca cctgggcttt
     3781 ctggatgtcc tctttaaagg tcaggctgtc gtcgtggatc agctgcatga agtttctgtt
     3841 ggcgaagccg tcggacttca ggaaatccag gattgtcttg ccggactgct tgtcccggat
     3901 gccgttgatc agcttccggc tcagcctgcc ccagccggtg tatctccgcc gcttcagctg
     3961 cttcatcact ttgtcgtcga acaggtgggc ataggttttc agccgttcct cgatcatctc
     4021 tctgtcctca aacagtgtca gggtcagcac gatatcttcc agaatgtcct cgttttcctc
     4081 attgtccagg aagtccttgt ccttgataat tttcagcaga tcgtggtatg tgcccaggga
     4141 ggcgttgaac cgatcttcca cgccggagat ttccacggag tcgaagcact cgattttctt
     4201 gaagtagtcc tctttcagct gcttcacggt cactttccgg ttggtcttga acagcaggtc
     4261 cacgatggcc tttttctgct cgccgctcag gaaggcgggc tttctcattc cctcggtcac
     4321 gtatttcact ttggtcagct cgttatacac ggtgaagtac tcgtacagca ggctgtgctt
     4381 gggcagcacc ttctcgttgg gcaggttctt atcgaagttg gtcatccgct cgatgaagct
     4441 ctgggcggaa gcgcccttgt ccaccacttc ctcgaagttc cagggggtga tggtttcctc
     4501 gctctttctg gtcatccagg cgaatctgct gtttcccctg gccagagggc ccacgtagta
     4561 ggggatgcgg aaggtcagga tcttctcgat cttttcccgg ttgtccttca ggaatgggta
     4621 aaaatcttcc tgccgccgca gaatggcgtg cagctctccc aggtggatct ggtgggggat
     4681 gctgccgttg tcgaaggtcc gctgcttccg cagcaggtcc tctctgttca gcttcacgag
     4741 cagttcctcg gtgccgtcca tcttttccag gatgggcttg atgaacttgt agaactcttc
     4801 ctggctggct ccgccgtcaa tgtagccggc gtagccgttc ttgctctggt cgaagaaaat
     4861 ctctttgtac ttctcaggca gctgctgccg cacgagagct ttcagcaggg tcaggtcctg
     4921 gtggtgctcg tcgtatctct tgatcataga ggcgctcagg ggggccttgg tgatctcggt
     4981 gttcactctc aggatgtcgc tcagcaggat ggcgtcggac aggttcttgg cggccagaaa
     5041 caggtcggcg tactggtcgc cgatctgggc cagcaggttg tccaggtcgt cgtcgtaggt
     5101 gtccttgctc agctgcagtt tggcatcctc ggccaggtcg aagttgctct tgaagttggg
     5161 ggtcaggccc aggctcaggg caatcaggtt tccgaacagg ccattcttct tctcgccggg
     5221 cagctgggcg atcagatttt ccagccgtct gctcttgctc agtctggcag acaggatggc
     5281 cttggcgtcc acgccgctgg cgttgatggg gttttcctcg aacagctggt tgtaggtctg
     5341 caccagctgg atgaacagct tgtccacgtc gctgttgtcg gggttcaggt cgccctcgat
     5401 caggaagtgg ccccggaact tgatcatgtg ggccagggcc agatagatca gccgcaggtc
     5461 ggccttgtcg gtgctgtcca ccagtttctt tctcaggtgg tagatggtgg ggtacttctc
     5521 gtggtaggcc acctcgtcca cgatgttgcc gaagatgggg tgccgctcgt gcttcttatc
     5581 ctcttccacc aggaaggact cttccagtct gtggaagaag ctgtcgtcca ccttggccat
     5641 ctcgttgctg aagatctctt gcagatagca gatccggttc ttccgtctgg tgtatcttct
     5701 tctggcggtt ctcttcagcc gggtggcctc ggctgtttcg ccgctgtcga acagcagggc
     5761 tccgatcagg ttcttcttga tgctgtgccg gtcggtgttg cccagcacct tgaatttctt
     5821 gctgggcacc ttgtactcgt cggtgatcac ggcccagccc acagagttgg tgccgatgtc
     5881 caggccgatg ctgtacttct tgtcggctgc tgggactccg tggataccga ccttccgctt
     5941 cttctttggg gccatcttat cgtcatcgtc tttgtaatcc atggtggcgg atcccgcgtc
     6001 acgacacctg tgttctggcg gcaaacccgt tgcgaaaaag aacgttcacg gcgactactg
     6061 cacttatata cggttctccc ccaccctcgg gaaaaaggcg gagccagtac acgacatcac
     6121 tttcccagtt taccccgcgc caccttctct aggcaccggt tcaattgccg acccctcccc
     6181 ccaacttctc ggggactgtg ggcgatgtgc gctctgccca ctgacgggca ccggagccac
     6241 tcgagccggc gccaaagtgg atctctgctg tccctgtaat aaacccgaaa attttgaatt
     6301 tttgtaattt gtttttgtaa ttctttagtt tgtatgtctg ttgctattat gtctactatt
     6361 ctttcccctg cactgtaccc cccaatcccc ccttttcttt taaaattgtg gatgaatact
     6421 gccatttgtc tcaagatcta gaattcaaaa aagcaccgac tcggtgccac tttttcaagt
     6481 tgataacgga ctagccttat tttaacttgc tatttctagc tctaaaacat cctgtcccta
     6541 gtggcccccg gtgtttcgtc ctttccacaa gatatataaa gccaagaaat cgaaatactt
     6601 tcaagttacg gtaagcatat gatagtccat tttaaaacat aattttaaaa ctgcaaacta
     6661 cccaagaaat tattactttc tacgtcacgt attttgtact aatatctttg tgtttacagt
     6721 caaattaatt ctaattatct ctctaacagc cttgtatcgt atatgcaaat atgaaggaat
     6781 catgggaaat aggccctctc tgggtcccct cggggttggg aggtgggtct gaaacgataa
     6841 tggtgaatat ccctgcctaa ctctattcac tatagaaagt acagcaaaaa ctattcttaa
     6901 acctaccaag cctcctacta tcattatgaa taattttata taccacagcc aatttgttat
     6961 gttaaaccaa ttccacaaac ttgcccattt atctaattcc aataattctt gttcattctt
     7021 ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt aagcttgtgt aattgttaat
     7081 ttctctgtcc cactccatcc aggtcgtgtg attccaaatc tgttccagag atttattact
     7141 ccaactagca ttccaaggca cagcagtggt gcaaatgagt tttccagagc aaccccaaat
     7201 ccccaggagc tgttgatcct ttaggtatct ttccacagcc aggattcttg cctggagctg
     7261 cttgatgccc cagactgtga gttgcaacag atgctgttgc gcctcaatag ccctcagcaa
     7321 attgttctgc tgctgcacta taccagacaa taattgtctg gcctgtaccg tcagcgtcat
     7381 tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac ccaaggaaca aagctcctat
     7441 tcccactgct cttttttctc tctgcaccac tcttctcttt gccttggtgg gtgctactcc
     7501 taatggttca atttttacta ctttatattt atataattca cttctccaat tgtccctcat
     7561 atctcctcct ccaggtctga agatcagcgg ccgcttgctg tgcggtggtc ttacttttgt
     7621 tttgctcttc ctctatcttg tctaaagctt ccttggtgtc ttttatctct atcctttgat
     7681 gcacacaata gagggttgct actgtattat ataatgatct aagttcttct gatcctgtct
     7741 gaagggatgg ttgtagctgt cccagtattt gtctacagcc ttctgatgtt tctaacaggc
     7801 caggattaac tgcgaatcgt tctagctccc tgcttgccca tactatatgt tttaatttat
     7861 attttttctt tccccctggc cttaaccgaa ttttttccca tcgcgatcta attctccccc
     7921 gcttaatact gacgctctcg cacccatctc tctccttcta gcctccgcta gtcaaaattt
     7981 ttggcgtact caccagtcgc cgcccctcgc ctcttgccgt gcgcgcttca gcaagccgag
     8041 tcctgcgtcg agagagctcc tctggtttcc ctttcgcttt caagtccctg ttcgggcgcc
     8101 actgctagag attttccaca ctgactaaaa gggtctgagg gatctctagt taccagagtc
     8161 acacaacaga cgggcacaca ctacttgaag cactcaaggc aagctttatt gaggcttaag
     8221 cagtgggttc cctagttagc cagagagctc ccaggctcag atctggtcta accagagaga
     8281 cccgtttatg tatcgagcta ggcacttaaa tacaatatct ctgcaatgcg gcaattcagt
     8341 ggttcgtcca atccatgtca gacccgtctg ttgccttcct aataaggcac gatcgtacca
     8401 ccttacttcc accaatcggc atgcacggtg ctttttctct ccttgtaagg catgttgcta
     8461 actcatcgtt accatgttgc aagactacaa gagtattgca taagactaca ttaagcttgc
     8521 agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     8581 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     8641 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     8701 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     8761 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     8821 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     8881 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     8941 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     9001 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     9061 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     9121 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     9181 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     9241 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     9301 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     9361 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     9421 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     9481 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     9541 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     9601 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     9661 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     9721 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     9781 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     9841 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     9901 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     9961 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
    10021 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
    10081 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
    10141 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
    10201 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
    10261 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
    10321 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
    10381 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
    10441 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
    10501 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
    10561 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
    10621 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
    10681 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
    10741 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
    10801 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
    10861 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
    10921 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
    10981 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
    11041 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
    11101 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
    11161 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
    11221 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
    11281 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
    11341 gtgagcgcgc gtaatacgac tcactatagg gcgaattggg tacgtcccta ggcctccaaa
    11401 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
    11461 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
    11521 cgggatgggc ggagttaggg gcgggatagc tagagccaga catgataaga tacattgatg
    11581 agtttggaca
//
