LOCUS       sequence-156364-       10679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  col1a1 targeting vector for inducible [TRE3G]-GFP-IRES-Cas9n (D10A 
            variant) expression. Contains NsiI cloning site for U6-sgRNA 
            cassettes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10679)
  AUTHORS   Dow LE, Fisher J, O'Rourke KP, Muley A, Kastenhuber ER, Livshits G, 
            Tschaharganeh DF, Socci ND, Lowe SW
  TITLE     Inducible in vivo genome editing with CRISPR-Cas9.
  JOURNAL   Nat Biotechnol. 2015 Apr;33(4):390-4. doi: 10.1038/nbt.3155. Epub 
            2015 Feb 18.
  PUBMED    25690852
REFERENCE   2  (bases 1 to 10679)
  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: "10.1038/nbt.3155"; 
            journalName: "Nat Biotechnol"; date: "2015-04"; volume: "33"; issue:
            "4"; pages: "390-4"
COMMENT     Sequence Label: c3GIC9n
FEATURES             Location/Qualifiers
     source          1..10679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..547
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(83..106)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     480..499
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     protein_bind    593..640
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    join(673..738,739..776)
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     polyA_signal    945..1079
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1199..1333
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1367..1745
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    1375..1393
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1411..1429
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1447..1465
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1483..1501
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1519..1537
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1555..1573
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1591..1609
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1654..1678
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(1769..1771,1772..1774,1775..2488)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2075..2094)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2422..2443
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2496..3063
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2513..3065
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2657..2674)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2884..2903
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3106..3132
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3136..3156
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3166..3189,3190..3192,3193..7266)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             3166..7266
                     /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
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7270..7290
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          7343..7915
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     rep_origin      complement(8243..8831)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8323..8342)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9002..9793,9794..9862))
                     /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     9625..9644
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9863..9967)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(9993..10448)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10130..10151)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10342..10361
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10575..10597
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10589..10606
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     10590..10606
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10616..10635
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        10616..10634
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     join(10667..10679,1..4)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(10667..10679,1..4)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 gatcccccgg gctgcaggaa ttaattcgtc gacctcgaaa ttctaccggg taggggaggc
       61 gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta
      121 cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc
      181 gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc
      241 cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct
      301 cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca
      361 atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg
      421 cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc
      481 attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg
      541 cctttcgacc tgcatccatc tagatctcga tcgagcagct gaagcttatg gagaagttac
      601 tattccgaag ttcctattct ctagaaagta taggaacttc gatccccctc gatcccccgg
      661 gctgcagatc tgtagggcgc agtagtccag ggtttccttg atgatgtcat acttatcctg
      721 tccctttttt ttccacagct cgcggttgag gacaaactct tcgcggtctt tccagtgggg
      781 atcgacggta tcgataagct tgcagatctg cgactctaga ggatctgcga ctctagagga
      841 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
      901 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
      961 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     1021 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct
     1081 gcgactctag aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     1141 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     1201 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     1261 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     1321 tcatgtctgg atcgagttta ccatgcatgt gttcgattct agattcgagt ttactcccta
     1381 tcagtgatag agaacgtatg aagagtttac tccctatcag tgatagagaa cgtatgcaga
     1441 ctttactccc tatcagtgat agagaacgta taaggagttt actccctatc agtgatagag
     1501 aacgtatgac cagtttactc cctatcagtg atagagaacg tatctacagt ttactcccta
     1561 tcagtgatag agaacgtata tccagtttac tccctatcag tgatagagaa cgtataagct
     1621 ttaggcgtgt acggtgggcg cctataaaag cagagctcgt ttagtgaacc gtcagatcgc
     1681 ctggagcaat tccacaacac ttttgtctta taccaacttt ccgtaccact tcctaccctc
     1741 gtaaagtcga gcttgcgttg gatccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     1801 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1861 tggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     1921 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     1981 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     2041 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     2101 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     2161 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     2221 ctatatcatg gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa
     2281 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2341 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     2401 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2461 tctcggcatg gacgagctgt acaagtaatg aattttccgc cccccccccc taacgttact
     2521 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
     2581 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
     2641 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
     2701 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
     2761 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
     2821 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
     2881 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
     2941 tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta
     3001 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     3061 taatatggcc ctcgagtatt gatcagtgga tccggtgcca ccatgtaccc atacgatgtt
     3121 ccagattacg cttcgccgaa gaaaaagcgc aaggtcgaag cgtccgacaa gaagtacagc
     3181 atcggcctgg ccatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag
     3241 gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac
     3301 ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga
     3361 accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc
     3421 agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg
     3481 gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg
     3541 gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc
     3601 gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc
     3661 cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc
     3721 cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg
     3781 gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc
     3841 gcccagctgc ccggcgagaa gaagaatggc ctgttcggca acctgattgc cctgagcctg
     3901 ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg
     3961 agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac
     4021 gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg
     4081 agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac
     4141 gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag
     4201 tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga
     4261 gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc
     4321 gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac
     4381 aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag
     4441 gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc
     4501 cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc
     4561 agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct
     4621 tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag
     4681 gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa
     4741 gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag
     4801 gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag
     4861 gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg
     4921 ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc
     4981 ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt
     5041 gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa
     5101 gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg
     5161 atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac
     5221 ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag
     5281 gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat
     5341 ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag
     5401 ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag
     5461 aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag
     5521 ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg
     5581 cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag
     5641 gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt
     5701 ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag
     5761 agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg
     5821 ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc
     5881 ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag
     5941 atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat
     6001 gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc
     6061 cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac
     6121 gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc
     6181 gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag
     6241 caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc
     6301 aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac
     6361 ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg
     6421 ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc
     6481 aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg
     6541 gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg
     6601 gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc
     6661 accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc
     6721 tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg
     6781 gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg
     6841 gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag
     6901 ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg
     6961 gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat
     7021 ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc
     7081 gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac
     7141 tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc
     7201 ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga
     7261 ggcgacagcc ccaagaagaa gagaaaggtg gaggccagct aagaattcgc cagcacagtg
     7321 ctcgagcgtt gagaacttca gggtgagttt ggggaccctt gattgttctt tctttttcgc
     7381 tattgtaaaa ttcatgttat atggaggggg caaagttttc agggtgttgt ttagaatggg
     7441 aagatgtccc ttgtatcacc atggaccctc atgataattt tgtttctttc actttctact
     7501 ctgttgacaa ccattgtctc ctcttatttt cttttcattt tctgtaactt tttcgttaaa
     7561 ctttagcttg catttgtaac gaatttttaa attcactttt gtttatttgt cagattgtaa
     7621 gtactttctc taatcacttt tttttcaagg caatcagggt atattatatt gtacttcagc
     7681 acagttttag agaacaattg ttataattaa atgataaggt agaatatttc tgcatataaa
     7741 ttctggctgg cgtggaaata ttcttattgg tagaaacaac tacaccctgg tcatcatcct
     7801 gcctttctct ttatggttac aatgatatac actgtttgag atgaggataa aatactctga
     7861 gtccaaaccg ggcccctctg ctaaccatgt tcatgccttc ttctctttcc tacagctcct
     7921 gggcaacgtg ctggttgttg tgctgtctca tcattttggc aaaggattca ctcctcaggt
     7981 gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca
     8041 ctgagatcgt tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct
     8101 gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga atttctcgat
     8161 cgctagcatc ctaggcgcgc cacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     8221 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     8281 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     8341 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     8401 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     8461 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     8521 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     8581 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     8641 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     8701 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     8761 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     8821 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     8881 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     8941 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     9001 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     9061 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     9121 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     9181 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     9241 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     9301 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     9361 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     9421 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     9481 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     9541 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     9601 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     9661 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     9721 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     9781 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     9841 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     9901 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     9961 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
    10021 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
    10081 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
    10141 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
    10201 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
    10261 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
    10321 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
    10381 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
    10441 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
    10501 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
    10561 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata
    10621 cgactcacta tagggcgaat tggagctcca ccgcggtggc ggccgctcta gaactagtg
//
