pST1374-NLS-flag-linker-Cas9

LOCUS       pST1374-NLS-flag-linker-Cas9                9317 bp ds-DNA     circular SYN 10-JAN-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pST1374-NLS-flag-linker-Cas9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9317)
  AUTHORS   Shen B, Zhang J, Wu H, Wang J, Ma K, Li Z, Zhang X, Zhang P, Huang X
  TITLE     Generation of gene-modified mice via Cas9/RNA-mediated gene 
            targeting.
  JOURNAL   Cell Res. 2013 Apr 2. doi: 10.1038/cr.2013.46.
  PUBMED    23545779
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|biofeng.com|
COMMENT     VNTNAME|pST1374-NLS-flag-linker-Cas9|
FEATURES             Location/Qualifiers
     source          1..9317
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    270..391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(281..300)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     335..354
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(549..947)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(966..1013)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(1061..1390)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1075..1210
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1129..1148)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1375..1395
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1404..1832)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1514..1535)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1726..1745
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    1878..2102
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2091..2108
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2131..2148)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2158..2199)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(2212..6315)
                     /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(6418..6441)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(6451..6471)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6499..6518)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6500..6518)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6563..6766)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6592..6612)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6767..7146
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     7318..7337
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7412..7516
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7517..8377
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7735..7754)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8548..9136
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9037..9056
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9290..9307
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
      241 agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac
      301 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      361 tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat
      421 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
      481 catagaaggc ggcggtggaa tcgaaatctc gtagcacgtg tcagtcctgc tcctcggcca
      541 cgaagtgctt agccctccca cacataacca gagggcagca attcacgaat cccaactgcc
      601 gtcggctgtc catcactgtc cttcactatg gctttgatcc caggatgcag atcgagaagc
      661 acctgtcggc accgtccgca ggggctcaag atgcccctgt tctcatttcc gatcgcgacg
      721 atacaagtca ggttgccagc tgccgcagca gcagcagtgc ccagcaccac gagttctgca
      781 caaggtcccc cagtaaaatg atatacattg acaccagtga agatgcggcc gtcgctagag
      841 agagctgcgc tggcgacgct gtagtcttca gagatgggga tgctgttgat tgtagccgtt
      901 gctctttcaa tgagggtgga ttcttcttga gacaaaggct tggccatggt ttagttcctc
      961 accttgtcgt attatactat gccgatatac tatgccgatg attaattgtc aacacgtgct
     1021 gatcagatcc gaaaatggat atacaagctc ccgggagctt tttgcaaaag cctaggcctc
     1081 caaaaaagcc tcctcactac ttctggaata gctcagaggc agaggcggcc tcggcctctg
     1141 cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta
     1201 ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc
     1261 tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat
     1321 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta
     1381 actgacacac attccacaga attaattcgc gttaaatttt tgttaaatca gctcattttt
     1441 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     1501 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     1561 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     1621 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     1681 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     1741 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     1801 cgccgcgctt aatgcgccgc tacagggcgc gtggggatac cccctagagc cccagctggt
     1861 tctttccgcc tcagaagcca tagagcccac cgcatcccca gcatgcctgc tattgtcttc
     1921 ccaatcctcc cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact
     1981 cagacaatgc gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc
     2041 cagggtcaag gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac
     2101 agtcgaggct gatcagcggg tttaaactca atggtgatgg tgatgatgac cggtacgcgt
     2161 agaatcgaga ccgaggagag ggttagggat aggcttacct tcgaagggcc cctagtcgcc
     2221 gccgagctga gacaggtcga tgcgagtctc gtacagtccg gtaattgact ggtgtatcag
     2281 ggtggcatcg agaacttctt tggtagaggt gtaccgcttc ctgtcaatag ttgtatcgaa
     2341 atacttgaag gcagcaggag cgcccagatt agtcagagta aagaggtgga taatattctc
     2401 tgcttgttcg cgaattggct tgtccctgtg cttattatat gcgctcagca ctttatcgag
     2461 gtttgcatcg gccagaataa cccgcttgct gaactcgcta atctgttcaa tgatttcgtc
     2521 caggtagtgt ttatgttgct caacaaagag ttgcttctgc tcattgtctt cagggctacc
     2581 tttgagtttc tcgtagtggg aggccagata caggaagttc acgtatttgg agggcagagc
     2641 cagctcgttt cctttctgca gctctccggc ggaggccagc atccgcttcc taccattctc
     2701 cagctcaaag agagagtact tgggcagttt gatgatgaga tctttcttca cttctttata
     2761 gcccttagct tccaggaaat cgattggatt cttctcgaag ctggatctct ccataatagt
     2821 aattccgagc agctccttaa cagacttgag tttcttggac ttgcctttct ccacttttgc
     2881 cacgaccaga acggaataag ccactgtagg ggaatcgaaa ccgccatact tctttgggtc
     2941 ccaatctttc ttcctggcga tcagcttgtc agagttccgc tttggcagga tgctctcctt
     3001 tgagaatccg ccggtctgca cttcggtctt cttcacgata ttgacttgtg gcatggacag
     3061 caccttccgc acagttgcga agtccctgcc tttatcccac acgatttctc ctgtttctcc
     3121 gtttgtttcg atcagtgggc gcttccggat ttcgccgtta gccagggtta tctcagtctt
     3181 aaagaaattc atgatattag agtagaagaa gtacttggcg gtggctttgc caatctcttg
     3241 ctcagacttt gctatcatct tcctcacatc gtagacttta tagtcaccgt acacgaactc
     3301 agactccagt ttagggtatt tcttgatcag ggcggtgcca acgacagcat tgagataggc
     3361 atcgtgagca tgatggtaat tgtttatttc gcgaactttg tagaattgaa agtccttccg
     3421 gaagtcagac accagcttgc tcttcagagt tatcactttc acttccctga tcagcttatc
     3481 gttctcatcg tacttagtgt tcatcctaga gtcgaggatt tgtgccacgt gtttggtaat
     3541 ctggcgtgtt tcgaccagtt gcctcttaat aaagccggcc ttgtcgagtt cgctcagacc
     3601 tcctctttct gccttagtca gattgtcaaa cttccgctgg gtgatcagtt tggcgttgag
     3661 gagctgccgc cagtaattct tcatcttctt gaccacctct tctgatggaa cattgtcaga
     3721 cttaccgcga ttcttatcgg atctggtcag caccttgttg tcaatggagt catctttgag
     3781 gaaggactgt ggaacaatat ggtccacgtc ataatcggac agccggttga tgtcgagttc
     3841 ctggtcaacg tacatgtccc gcccgttctg gaggtagtac aggtagagtt tctcgttctg
     3901 gagctgtgta ttctccacag ggtgctcctt cagtatctga gatcccagct ccttaattcc
     3961 ctcttcgatt cttttcatcc gttcccgaga gttcttctgg cccttctggg tggtttggtt
     4021 ctcccttgcc atttcgataa cgatattctc tggcttgtgc cgacccatca ctttgacgag
     4081 ttcgtccacg accttgactg tctgcagtat tcccttcttt atggcgggtg atccagcgag
     4141 gttggcgatg tgctcgtgca ggctgtcgcc ttgaccgctc acctgtgcct tctggatgtc
     4201 ctctttaaat gtcagagagt catcgtgaat cagttgcatg aagttcctgt tagcgaatcc
     4261 gtcggatttc aggaaatcca ggatggtctt tccgctctgt ttgtcgcgga tgccgtttat
     4321 gagtttcctg gagagtctac cccacccagt gtagcgccgc cgcttgagct gtttcatgac
     4381 tttatcgtca aacagatggg cataggtctt caggcgctct tcgatcatct ctctatcctc
     4441 gaacagagtc agggtcagca cgatatcttc caggatgtcc tcgttctcct cattatcgag
     4501 gaaatctttg tctttgatta tcttcagcag atcatggtaa gtgcccaggc tggcattaaa
     4561 gcggtcctcc acgccagaga tttccactga gtcaaagcat tcgatcttct taaagtaatc
     4621 ctccttgagc tgcttcactg tcacctttct attagtcttg aagagcagat caacgatagc
     4681 cttcttctgc tctccggaca ggaaggcagg cttcctcatg ccctcggtca cgtacttcac
     4741 tttagtcagc tcgttataga cggtgaaata ctcgtacagg agtgaatgtt tgggcaggac
     4801 cttctcgttg ggcaggttct tatcgaaatt ggtcatccgt tcgatgaatg actgggcgct
     4861 tgcgccctta tccacgacct cttcgaagtt ccaaggagta attgtctcct cagatttcct
     4921 ggtcatccaa gcgaagcggg agttgcctct agccagaggg ccgacgtaat aagggatcct
     4981 gaaggtcagg atcttctcta tcttctcccg gttatccttc aggaaaggat agaaatcctc
     5041 ctgcctgcgg aggattgcat gcagctctcc caggtgtatc tgatgtggaa tggagccatt
     5101 atcaaaggtc ctctgcttcc tcagcaggtc ctccctgttc agcttcacca gcagctcttc
     5161 agtaccgtcc atcttctcga ggattggttt gatgaacttg taaaattctt cctgtgatgc
     5221 tccgccatcg atgtatccgg catatccatt cttgctctgg tcgaagaata tctctttgta
     5281 cttctctggc agctgttgcc tcacgagggc tttgagcaga gtcaggtctt gatggtgttc
     5341 atcatagcgt tttatcatgg aggcgctcag aggtgctttg gtgatctcag tgttgacccg
     5401 gagtatgtcg ctcagcagaa tggcgtcgga gagattctta gcagccagaa agagatcggc
     5461 gtactggtcg cctatctgtg cgagcaggtt gtccagatca tcgtcatagg tgtccttgga
     5521 gagctggagt ttagcatctt cggccagatc aaaattggac ttgaagttag gtgtcaggcc
     5581 caggctcagg gcgatgaggt tcccaaacag gccgttcttc ttttctcctg gcagttgggc
     5641 aatcagattc tccagtctgc gtgacttgga cagccgagcg gacagaatag ccttggcatc
     5701 cacaccagaa gcgtttatgg gattctcctc gaacagttgg ttatatgtct gcaccagttg
     5761 aatgaagagt ttatccacat cggaattatc gggattcagg tcgccctcga tcagaaagtg
     5821 tcctctaaac tttatcatat gagccagggc cagatagatg agcctcaggt ctgctttatc
     5881 ggtgctgtcc accagcttct tcctcagatg gtagattgta ggatactttt catggtaagc
     5941 cacctcatcc acgatatttc cgaatatagg gtgcctctcg tgtttcttat cctcctccac
     6001 cagaaagctc tcttccaggc ggtgaaagaa ggagtcgtcc accttagcca tttcgttgct
     6061 aaagatctct tgcagataac atatccgatt cttgcggcgg gtgtatctcc gccttgcggt
     6121 ccgcttcagc cgagtagctt cagcggtttc accggagtcg aagaggagtg ctccgatcag
     6181 gttcttcttg attgaatggc ggtcagtatt acccagcacc ttgaatttct tgcttggcac
     6241 cttatactcg tcggttatga cggcccagcc aacggagttt gtcccgatat ccagtccaat
     6301 agagtatttc ttgtctctag tgtgggtccg ctggtgtctg gtcagagcac cagactgaga
     6361 gaaagattta ccacactctg ggcatttgta tggcttctcg ccgggttcca gtctagattt
     6421 atcgtcgtca tccttgtagt cagcggccgc caccttcctc tttttcttag gtcccatggt
     6481 gctagccagc ttgggtctcc ctatagtgag tcgtattaat ttcgataagc cagtaagcag
     6541 tgggttctct agttagccag agagctctgc ttatatagac ctcccaccgt acacgcctac
     6601 cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt
     6661 ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt
     6721 caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga
     6781 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
     6841 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     6901 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
     6961 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg
     7021 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca
     7081 tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta gtcaataatc
     7141 aatgtcaacg cgtatatctg gcccgtacat cgcgaagcag cgcaaaacgc ctaaccctaa
     7201 gcagattctt catgcaattg tcggtcaagc cttgccttgt tgtagcttaa attttgctcg
     7261 cgcactactc agcgacctcc aacacacaag cagggagcag atactggctt aactatgcgg
     7321 catcagagca gattgtactg agagtgcacc ataggggatc gggagatctc ccgatccgtc
     7381 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     7441 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     7501 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     7561 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     7621 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     7681 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     7741 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     7801 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     7861 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     7921 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     7981 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     8041 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     8101 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     8161 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     8221 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     8281 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     8341 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     8401 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     8461 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     8521 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     8581 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     8641 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     8701 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     8761 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     8821 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     8881 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     8941 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     9001 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     9061 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     9121 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     9181 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     9241 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     9301 gagcgaggaa gcggaag
//