LOCUS       sequence-497737-       14538 bp    DNA     circular SYN 16-JAN-2026
DEFINITION  Lentiviral vector for stable expression of dCas9-3XGFP-CRY2hiclu 
            fusion protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14538)
  AUTHORS   Stortz M, Keikhosravi A, Pegoraro G, Misteli T
  TITLE     OptoLoop: An optogenetic tool to probe the functional role of genome
            organization
  JOURNAL   bioRxiv 2025.11.06.686574
REFERENCE   2  (bases 1 to 14538)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 16, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2025.11.06.686574"
COMMENT     Sequence Label: pHAGE-dCas9-3XGFP-CRY2
FEATURES             Location/Qualifiers
     source          1..14538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2299..2602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2603..2806
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2757..2777
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2808..2826
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2829..2847
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2857..2881
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2974..2983
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2986..3006
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3025..3048,3049..3051,3052..5535,5536..5541,5542..5607,
                     5608..5610,5611..7125)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(3025..3048,3049..3051,3052..5538,5539..5541,
                     5542..7125)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             3025..7125
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7144..7164
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7177..7890
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             7909..8622
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             8641..9354
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             9388..10881
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTRRAQIMIGAA"
     misc_feature    10928..11516
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10981..11001)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11399..11410)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11591..11824
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(11943..11961)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        12029..12133
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(12134..12202,12203..12994)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(12352..12371)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      13165..13753
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13654..13673
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13907..13924
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    14164..14185
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(14200..14205,14206..14223,14224..14230)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14238..14254
                     /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     14243..14265
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     14262..14278
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     14262..14278
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     14328..14347
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttgacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagcgat
     2221 gctagctgat gcgggcccta gttattaata gtaatcaatt acggggtcat tagttcatag
     2281 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     2341 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     2401 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     2461 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     2521 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     2581 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     2641 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     2701 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     2761 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctcc ctatcagtga
     2821 tagagatctc cctatcagtg atagagatcg tcgacgagct cgtttagtga accgtcagat
     2881 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
     2941 cctaccggtt aaggtgaacg cgtgcaggct ggcgccacca tggcccctaa aaagaaacgc
     3001 aaggtcgaat tcggaagcgg aagcgacaag aagtactcca ttgggctcgc tatcggtacc
     3061 aacagcgtcg gctgggccgt cattacggac gagtacaagg tgccgagcaa aaaattcaaa
     3121 gttctgggca ataccgatcg ccacagcata aagaagaacc tcattggagc cctcctgttc
     3181 gactccgggg agacggccga agccacgcgg ctcaaaagaa cagcacggcg cagatatacc
     3241 cgcagaaaga atcggatctg ctacctgcag gagatcttta gtaatgagat ggctaaggtg
     3301 gatgactctt tcttccatag gctggaggag tcctttttgg tggaggagga taaaaagcac
     3361 gagcgccacc caatctttgg caatatcgtg gacgaggtgg cgtaccatga aaagtaccca
     3421 accatatatc atctgaggaa gaagctggta gacagtactg ataaggctga cttgcggttg
     3481 atctatctcg cgctggcgca catgatcaaa tttcggggac acttcctcat cgagggggac
     3541 ctgaacccag acaacagcga tgtcgacaaa ctctttatcc aactggttca gacttacaat
     3601 cagcttttcg aggagaaccc gatcaacgca tccggcgttg acgccaaagc aatcctgagc
     3661 gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg cacagctccc tggggagaag
     3721 aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc caactttaaa
     3781 tctaacttcg acctggccga agatgccaag ctgcaactga gcaaagacac ctacgatgat
     3841 gatctcgaca atctgctggc ccagatcggc gaccagtacg cagacctttt tttggcggca
     3901 aagaacctgt cagacgccat tctgctgagt gatattctgc gagtgaacac ggagatcacc
     3961 aaagctccgc tgagcgctag tatgatcaag cgctatgatg agcaccacca agacttgact
     4021 ttgctgaagg cccttgtcag acagcaactg cctgagaagt acaaggaaat tttcttcgat
     4081 cagtctaaaa atggctacgc cggatacatt gacggcggag caagccagga ggaattttac
     4141 aaatttatta agcccatctt ggaaaaaatg gacggcaccg aggagctgct ggtaaagctg
     4201 aacagagaag atctgttgcg caaacagcgc actttcgaca atggaagcat cccccaccag
     4261 attcacctgg gcgaactgca cgctatcctc aggcggcaag aggatttcta cccctttttg
     4321 aaagataaca gggaaaagat tgagaaaatc ctcacatttc ggatacccta ctatgtaggc
     4381 cccctcgctc ggggaaattc cagattcgcg tggatgactc gcaaatcaga agagaccatc
     4441 actccctgga acttcgagga agtcgtggat aagggggcct ctgcccagtc cttcatcgaa
     4501 aggatgacta actttgataa aaatctgcct aacgaaaagg tgcttcctaa acactctctg
     4561 ctgtacgagt acttcacagt ttataacgag ctcaccaagg tcaaatacgt cacagaaggg
     4621 atgagaaagc cagcattcct gtctggagag cagaagaaag ctatcgtgga cctcctcttc
     4681 aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag actatttcaa aaagattgaa
     4741 tgtttcgact ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc cctgggaacg
     4801 tatcacgatc tcctgaaaat cattaaagac aaggacttcc tggacaatga ggagaacgag
     4861 gacattcttg aggacattgt cctcaccctt acgttgtttg aagataggga gatgattgaa
     4921 gaacgcttga aaacttacgc tcatctcttc gacgacaaag tcatgaaaca gctcaagaga
     4981 cgccgatata caggatgggg gcggctgtca agaaaactga tcaatggcat ccgagacaag
     5041 cagagtggaa agacaatcct ggattttctt aagtccgatg gatttgccaa ccggaacttc
     5101 atgcagttga tccatgatga ctctctcacc tttaaggagg acatccagaa agcacaagtt
     5161 tctggccagg gggacagtct tcacgagcac atcgctaatc ttgcaggtag cccagctatc
     5221 aaaaagggaa tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt aatgggaagg
     5281 cataagcccg agaatatcgt tatcgagatg gcccgagaga accaaactac ccagaaggga
     5341 cagaagaaca gtagggaaag gatgaagagg attgaagagg gtataaaaga actggggtcc
     5401 caaatcctta aggaacaccc agttgaaaac acccagcttc agaatgagaa gctctacctg
     5461 tactacctgc agaacggcag ggacatgtac gtggatcagg aactggacat caaccggttg
     5521 tccgactacg acgtggatgc tatcgtgccc caaagctttc tcaaagatga ttctattgat
     5581 aataaagtgt tgacaagatc cgataaaaat agagggaaga gtgataacgt cccctcagaa
     5641 gaagttgtca agaaaatgaa aaattattgg cggcagctgc tgaacgccaa actgatcaca
     5701 caacggaagt tcgataatct gactaaggct gaacgaggtg gcctgtctga gttggataaa
     5761 gccggcttca tcaaaaggca gcttgttgag acacgccaga tcaccaagca cgtggcccaa
     5821 attctcgatt cacgcatgaa caccaagtac gatgaaaatg acaaactgat tcgagaggtg
     5881 aaagttatta ctctgaagtc taagctggtc tcagatttca gaaaggactt tcagttttat
     5941 aaggtgagag agatcaacaa ttaccaccat gcgcatgatg cctacctgaa tgcagtggta
     6001 ggcactgcac ttatcaaaaa atatcccaag ctggaatctg aatttgttta cggagactat
     6061 aaagtgtacg atgttaggaa aatgatcgca aagtctgagc aggaaatagg caaggccacc
     6121 gctaagtact tcttttacag caatattatg aattttttca agaccgagat tacactggcc
     6181 aatggagaga ttcggaagcg accacttatc gaaacaaacg gagaaacagg agaaatcgtg
     6241 tgggacaagg gtagggattt cgcgacagtc cgcaaggtcc tgtccatgcc gcaggtgaac
     6301 atcgttaaaa agaccgaagt acagaccgga ggcttctcca aggaaagtat cctcccgaaa
     6361 aggaacagcg acaagctgat cgcacgcaaa aaagattggg accccaagaa atacggcgga
     6421 ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga gaaagggaag
     6481 tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca caatcatgga gcgatccagc
     6541 ttcgagaaaa accccatcga ctttctcgaa gcgaaaggat ataaagaggt caaaaaagac
     6601 ctcatcatta agctgcccaa gtactctctc tttgagcttg aaaacggccg gaaacgaatg
     6661 ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg cactgccctc taaatacgtt
     6721 aatttcttgt atctggccag ccactatgaa aagctcaaag ggtctcccga agataatgag
     6781 cagaagcagc tgttcgtgga acaacacaaa cactaccttg atgagatcat cgagcaaata
     6841 agcgagttct ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt gctttctgct
     6901 tacaataagc acagggataa gcccatcagg gagcaggcag aaaacattat ccacttgttt
     6961 actctgacca acttgggcgc gcctgcagcc ttcaagtact tcgacaccac catagacaga
     7021 aagcggtaca cctctacaaa ggaggtcctg gacgccacac tgattcatca gtcaattacg
     7081 gggctctatg aaacaagaat cgacctctct cagctcggtg gagacggctc tactagtggc
     7141 tctcccaaga agaagaggaa ggtaggatcc ggaagtatgc gtaaaggcga agagctgttc
     7201 actggtgtcg tccctattct ggtggaactg gatggtgatg tcaacggtca taagttttcc
     7261 gtgcgtggcg agggtgaagg tgacgcaact aatggtaaac tgacgctgaa gttcatctgt
     7321 actactggta aactgccggt accttggccg actctggtaa cgacgctgac ttatggtgtt
     7381 cagtgctttg ctcgttatcc ggaccatatg aagcagcatg acttcttcaa gtccgccatg
     7441 ccggaaggct atgtgcagga acgcacgatt tcctttaagg atgacggcac gtacaaaacg
     7501 cgtgcggaag tgaaatttga aggcgatacc ctggtaaacc gcattgagct gaaaggcatt
     7561 gactttaaag aagacggcaa tatcctgggc cataagctgg aatacaattt taacagccac
     7621 aatgtttaca tcaccgccga taaacaaaaa aatggcatta aagcgaattt taaaattcgc
     7681 cacaacgtgg aggatggcag cgtgcagctg gctgatcact accagcaaaa cactccaatc
     7741 ggtgatggtc ctgttctgct gccagacaat cactatctga gcacgcaaag cgttctgtct
     7801 aaagatccga acgagaaacg cgatcatatg gttctgctgg agttcgtaac cgcagcgggc
     7861 atcacgcatg gtatggatga actgtacaaa tctggcggcc gctctggcat gcgtaaaggc
     7921 gaagagctgt tcactggtgt cgtccctatt ctggtggaac tggatggtga tgtcaacggt
     7981 cataagtttt ccgtgcgtgg cgagggtgaa ggtgacgcaa ctaatggtaa actgacgctg
     8041 aagttcatct gtactactgg taaactgccg gtaccttggc cgactctggt aacgacgctg
     8101 acttatggtg ttcagtgctt tgctcgttat ccggaccata tgaagcagca tgacttcttc
     8161 aagtccgcca tgccggaagg ctatgtgcag gaacgcacga tttcctttaa ggatgacggc
     8221 acgtacaaaa cgcgtgcgga agtgaaattt gaaggcgata ccctggtaaa ccgcattgag
     8281 ctgaaaggca ttgactttaa agaagacggc aatatcctgg gccataagct ggaatacaat
     8341 tttaacagcc acaatgttta catcaccgcc gataaacaaa aaaatggcat taaagcgaat
     8401 tttaaaattc gccacaacgt ggaggatggc agcgtgcagc tggctgatca ctaccagcaa
     8461 aacactccaa tcggtgatgg tcctgttctg ctgccagaca atcactatct gagcacgcaa
     8521 agcgttctgt ctaaagatcc gaacgagaaa cgcgatcata tggttctgct ggagttcgta
     8581 accgcagcgg gcatcacgca tggtatggat gaactgtaca aaggtagcat cgatggtagc
     8641 atgcgtaaag gcgaagagct gttcactggt gtcgtcccta ttctggtgga actggatggt
     8701 gatgtcaacg gtcataagtt ttccgtgcgt ggcgagggtg aaggtgacgc aactaatggt
     8761 aaactgacgc tgaagttcat ctgtactact ggtaaactgc cggtaccttg gccgactctg
     8821 gtaacgacgc tgacttatgg tgttcagtgc tttgctcgtt atccggacca tatgaagcag
     8881 catgacttct tcaagtccgc catgccggaa ggctatgtgc aggaacgcac gatttccttt
     8941 aaggatgacg gcacgtacaa aacgcgtgcg gaagtgaaat ttgaaggcga taccctggta
     9001 aaccgcattg agctgaaagg cattgacttt aaagaagacg gcaatatcct gggccataag
     9061 ctggaataca attttaacag ccacaatgtt tacatcaccg ccgataaaca aaaaaatggc
     9121 attaaagcga attttaaaat tcgccacaac gtggaggatg gcagcgtgca gctggctgat
     9181 cactaccagc aaaacactcc aatcggtgat ggtcctgttc tgctgccaga caatcactat
     9241 ctgagcacgc aaagcgttct gtctaaagat ccgaacgaga aacgcgatca tatggttctg
     9301 ctggagttcg taaccgcagc gggcatcacg catggtatgg atgaactgta caaacctgga
     9361 ggttctggcg gccgcggtgg agccaccatg aagatggaca aaaagactat agtttggttt
     9421 agaagagacc taaggattga ggataatcct gcattagcag cagctgctca cgaaggatct
     9481 gtttttcctg tcttcatttg gtgtcctgaa gaagaaggac agttttatcc tggaagagct
     9541 tcaagatggt ggatgaaaca atcacttgct cacttatctc aatccttgaa ggctcttgga
     9601 tctgacctca ctttaatcaa aacccacaac acgatttcag cgatcttgga ttgtatccgc
     9661 gttaccggtg ctacaaaagt cgtctttaac cacctctatg atcctgtttc gttagttcgg
     9721 gaccataccg taaaggagaa gctggtggaa cgtgggatct ctgtgcaaag ctacaatgga
     9781 gatctattgt atgaaccgtg ggagatatac tgcgaaaagg gcaaaccttt tacgagtttc
     9841 aattcttact ggaagaaatg cttagatatg tcgattgaat ccgttatgct tcctcctcct
     9901 tggcggttga tgccaataac tgcagcggct gaagcgattt gggcgtgttc gattgaagaa
     9961 ctagggctgg agaatgaggc cgagaaaccg agcaatgcgt tgttaactag agcttggtct
    10021 ccaggatgga gcaatgctga taagttacta aatgagttca tcgagaagca gttgatagat
    10081 tatgcaaaga acagcaagaa agttgttggg aattctactt cactactttc tccgtatctc
    10141 catttcgggg aaataagcgt cagacacgtt ttccagtgtg cccggatgaa acaaattata
    10201 tgggcaagag ataagaacag tgaaggagaa gaaagtgcag atctttttct taggggaatc
    10261 ggtttaagag agtattctcg gtatatatgt ttcaacttcc cgtttactca cgagcaatcg
    10321 ttgttgagtc atcttcggtt tttcccttgg gatgctgatg ttgataagtt caaggcctgg
    10381 agacaaggca ggaccggtta tccgttggtg gatgccggaa tgagagagct ttgggctacc
    10441 ggatggatgc ataacagaat aagagtgatt gtttcaagct ttgctgtgaa gtttcttctc
    10501 cttccatgga aatggggaat gaagtatttc tgggatacac ttttggatgc tgatttggaa
    10561 tgtgacatcc ttggctggca gtatatctct gggagtatcc ccgatggcca cgagcttgat
    10621 cgcttggaca atcccgcgtt acaaggcgcc aaatatgacc cagaaggtga gtacataagg
    10681 caatggcttc ccgagcttgc gagattgcca actgaatgga tccatcatcc atgggacgct
    10741 cctttaaccg tactcaaagc ttctggtgtg gaactcggaa caaactatgc gaaacccatt
    10801 gtagacatcg acacagctcg tgagctacta gctaaagcta tttcaagaac ccgtagagca
    10861 cagatcatga tcggagcagc agcccgggat ccaccggacc tcgacaacta agtagggtaa
    10921 tctagataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
    10981 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
    11041 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
    11101 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
    11161 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
    11221 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
    11281 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
    11341 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
    11401 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
    11461 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctga
    11521 gatcctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
    11581 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
    11641 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
    11701 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
    11761 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
    11821 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
    11881 tcagagagtg agaggcccgg gttaattaag gaaagggcta gatcattctt gaagacgaaa
    11941 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
    12001 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
    12061 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
    12121 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
    12181 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
    12241 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
    12301 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
    12361 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
    12421 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
    12481 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
    12541 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
    12601 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
    12661 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
    12721 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
    12781 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
    12841 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
    12901 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
    12961 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
    13021 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
    13081 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
    13141 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
    13201 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
    13261 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
    13321 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
    13381 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
    13441 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
    13501 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
    13561 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
    13621 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
    13681 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
    13741 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
    13801 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
    13861 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
    13921 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
    13981 ttaatgcagc aagctcatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
    14041 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
    14101 aaaaagctcc ccgtggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
    14161 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
    14221 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacat
    14281 gattacgaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
    14341 actcatcaat gtatcttatc atgtctggat caactggata actcaagcta accaaaatca
    14401 tcccaaactt cccaccccat accctattac cactgccaat tacctgtggt ttcatttact
    14461 ctaaacctgt gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg
    14521 tactacaaac ttagtagt
//
