LOCUS       Exported               10751 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  col1a1 targeting vector for inducible [TRE3G]-GFP-IRES-Cas9 
            expression. Contains NsiI cloning site for U6-sgRNA cassettes.
ACCESSION   .
VERSION     .
KEYWORDS    c3GIC9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10751)
  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 10751)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10751
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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    673..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             1769..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(1814..1835)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     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    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"
     regulatory      3097..3106
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3106..3171
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3178..3198
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3223..7323
                     /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             7324..7371
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     intron          7415..7987
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     rep_origin      complement(8315..8903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8395..8414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9074..9934)
                     /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     9697..9716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9935..10039)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10065..10520)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10202..10223)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10414..10433
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10647..10669
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10661..10678
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     10662..10678
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10688..10707
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        10688..10706
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     10739..4
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10739..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 ccatggacta taaggaccac
     3121 gacggagact acaaggatca tgatattgat tacaaagacg atgacgataa gatggcccca
     3181 aagaagaagc ggaaggtcgg tatccacgga gtcccagcag ccgacaagaa gtacagcatc
     3241 ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg
     3301 cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg
     3361 atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc
     3421 gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc
     3481 aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg
     3541 gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc
     3601 taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac
     3661 aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac
     3721 ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag
     3781 ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac
     3841 gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc
     3901 cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct gagcctgggc
     3961 ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc
     4021 aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc
     4081 gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga
     4141 gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag
     4201 caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac
     4261 aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga cggcggagcc
     4321 agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag
     4381 gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac
     4441 ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa
     4501 gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc
     4561 atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga
     4621 aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgcttcc
     4681 gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg
     4741 ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg
     4801 aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaggcc
     4861 atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac
     4921 tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc
     4981 aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg
     5041 gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag
     5101 gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg
     5161 atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc
     5221 aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc
     5281 ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac
     5341 atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg
     5401 gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc
     5461 gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac
     5521 cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc
     5581 atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag
     5641 aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa
     5701 ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca gagctttctg
     5761 aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg gggcaagagc
     5821 gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg gcagctgctg
     5881 aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga gagaggcggc
     5941 ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc
     6001 acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaatgac
     6061 aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg
     6121 aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc
     6181 tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcgag
     6241 ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag
     6301 gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag
     6361 accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc
     6421 gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg gaaagtgctg
     6481 agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa
     6541 gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa ggactgggac
     6601 cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc
     6661 aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc
     6721 atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac
     6781 aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa
     6841 aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc
     6901 ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc
     6961 tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac
     7021 gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg
     7081 gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag
     7141 aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt
     7201 gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg
     7261 atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggc
     7321 gacaaaaggc cggcggccac gaaaaaggcc ggccaggcaa aaaagaaaaa gtaagaattc
     7381 gccagcacag tgctcgagcg ttgagaactt cagggtgagt ttggggaccc ttgattgttc
     7441 tttctttttc gctattgtaa aattcatgtt atatggaggg ggcaaagttt tcagggtgtt
     7501 gtttagaatg ggaagatgtc ccttgtatca ccatggaccc tcatgataat tttgtttctt
     7561 tcactttcta ctctgttgac aaccattgtc tcctcttatt ttcttttcat tttctgtaac
     7621 tttttcgtta aactttagct tgcatttgta acgaattttt aaattcactt ttgtttattt
     7681 gtcagattgt aagtactttc tctaatcact tttttttcaa ggcaatcagg gtatattata
     7741 ttgtacttca gcacagtttt agagaacaat tgttataatt aaatgataag gtagaatatt
     7801 tctgcatata aattctggct ggcgtggaaa tattcttatt ggtagaaaca actacaccct
     7861 ggtcatcatc ctgcctttct ctttatggtt acaatgatat acactgtttg agatgaggat
     7921 aaaatactct gagtccaaac cgggcccctc tgctaaccat gttcatgcct tcttctcttt
     7981 cctacagctc ctgggcaacg tgctggttgt tgtgctgtct catcattttg gcaaaggatt
     8041 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     8101 tcacaaatac cactgagatc gttttccctc tgccaaaaat tatggggaca tcatgaagcc
     8161 ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     8221 gaatttctcg atcgctagca tcctaggcgc gccacatgtg agcaaaaggc cagcaaaagg
     8281 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     8341 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     8401 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     8461 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     8521 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     8581 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     8641 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     8701 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     8761 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     8821 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     8881 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     8941 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     9001 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     9061 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     9121 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     9181 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     9241 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     9301 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     9361 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     9421 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     9481 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     9541 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     9601 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     9661 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     9721 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     9781 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     9841 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     9901 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     9961 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
    10021 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
    10081 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
    10141 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
    10201 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
    10261 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
    10321 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
    10381 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
    10441 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
    10501 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
    10561 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
    10621 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
    10681 gcgcgcgtaa tacgactcac tatagggcga attggagctc caccgcggtg gcggccgctc
    10741 tagaactagt g
//
