LOCUS       Exported               11169 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expression dzCas9-KRAB in microalgae. CRISPR/Cas based plant genome 
            editing and gene regulation, Hyg resistance.
ACCESSION   .
VERSION     .
KEYWORDS    pHdzCas9-KRAB
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11169)
  AUTHORS   Kao PH, Ng IS
  TITLE     CRISPRi mediated phosphoenolpyruvate carboxylase regulation to 
            enhance the production of lipid in Chlamydomonas reinhardtii.
  JOURNAL   Bioresour Technol. 2017 May 4. pii: S0960-8524(17)30619-3. doi: 
            10.1016/j.biortech.2017.04.111.
  PUBMED    28501380
REFERENCE   2  (bases 1 to 11169)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11169
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..345
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             528..593
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             600..620
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             645..4745
                     /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"
     RBS             4658..4666
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             4746..4793
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4812..4832
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4848..4868
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4905..5090
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             5097..5117
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5124..5150
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      5203..5455
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        5572..6248
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             6315..7340
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    7381..7555
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    7705..7729
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(7977..8000)
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     terminator      8008..8035
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      8127..8213
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      8382..8817
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     rep_origin      complement(8932..9520)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9012..9031)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9610..10419)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     10333..10352
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     10411..10430
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     misc_feature    10648..10672
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 tgagactttt caacaaaggg taatatccgg aaacctcctc ggattccatt gcccagctat
       61 ctgtcacttt attgtgaaga tagtggaaaa ggaaggtggc tcctacaaat gccatcattg
      121 cgataaagga aaggccatcg ttgaagatgc ctctgccgac agtggtccca aagatggacc
      181 cccacccacg aggagcatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt
      241 ggattgatgt gatatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca
      301 agacccttcc tctatataag gaagttcatt tcatttggag aggacctcga cctcaacaca
      361 acatatacaa aacaaacgaa tctcaagcaa tcaagcattc tacttctatt gcagcaattt
      421 aaatcatttc ttttaaagca aaagcaattt tctgaaaatt ttcaccattt acgaacgata
      481 ctcgaggggg atccccaata cttgtatggc cgcggccgct ctagatggat tacaaggacc
      541 acgacgggga ttacaaggac cacgacattg attacaagga tgatgatgac aagatggctc
      601 cgaagaagaa gaggaaggtt ggcatccacg gggtgccagc tgctgacaag aagtactcga
      661 tcggcctcgc cattgggact aactctgttg gctgggccgt gatcaccgac gagtacaagg
      721 tgccctcaaa gaagttcaag gtcctgggca acaccgatcg gcattccatc aagaagaatc
      781 tcattggcgc tctcctgttc gacagcggcg agacggctga ggctacgcgg ctcaagcgca
      841 ccgcccgcag gcggtacacg cgcaggaaga atcgcatctg ctacctgcag gagattttct
      901 ccaacgagat ggcgaaggtt gacgattctt tcttccacag gctggaggag tcattcctcg
      961 tggaggagga taagaagcac gagcggcatc caatcttcgg caacattgtc gacgaggttg
     1021 cctaccacga gaagtaccct acgatctacc atctgcggaa gaagctcgtg gactccacag
     1081 ataaggcgga cctccgcctg atctacctcg ctctggccca catgattaag ttcaggggcc
     1141 atttcctgat cgagggggat ctcaacccgg acaatagcga tgttgacaag ctgttcatcc
     1201 agctcgtgca gacgtacaac cagctcttcg aggagaaccc cattaatgcg tcaggcgtcg
     1261 acgcgaaggc tatcctgtcc gctaggctct cgaagtctcg gcgcctcgag aacctgatcg
     1321 cccagctgcc gggcgagaag aagaacggcc tgttcgggaa tctcattgcg ctcagcctgg
     1381 ggctcacgcc caacttcaag tcgaatttcg atctcgctga ggacgccaag ctgcagctct
     1441 ccaaggacac atacgacgat gacctggata acctcctggc ccagatcggc gatcagtacg
     1501 cggacctgtt cctcgctgcc aagaatctgt cggacgccat cctcctgtct gatattctca
     1561 gggtgaacac cgagattacg aaggctccgc tctcagcctc catgatcaag cgctacgacg
     1621 agcaccatca ggatctgacc ctcctgaagg cgctggtcag gcagcagctc cccgagaagt
     1681 acaaggagat cttcttcgat cagtcgaaga acggctacgc tgggtacatt gacggcgggg
     1741 cctctcagga ggagttctac aagttcatca agccgattct ggagaagatg gacggcacgg
     1801 aggagctgct ggtgaagctc aatcgcgagg acctcctgag gaagcagcgg acattcgata
     1861 acggcagcat cccacaccag attcatctcg gggagctgca cgctatcctg aggaggcagg
     1921 aggacttcta ccctttcctc aaggataacc gcgagaagat cgagaagatt ctgactttca
     1981 ggatcccgta ctacgtcggc ccactcgcta ggggcaactc ccgcttcgct tggatgaccc
     2041 gcaagtcaga ggagacgatc acgccgtgga acttcgagga ggtggtcgac aagggcgcta
     2101 gcgctcagtc gttcatcgag aggatgacga atttcgacaa gaacctgcca aatgagaagg
     2161 tgctccctaa gcactcgctc ctgtacgagt acttcacagt ctacaacgag ctgactaagg
     2221 tgaagtatgt gaccgagggc atgaggaagc cggctttcct gtctggggag cagaagaagg
     2281 ccatcgtgga cctcctgttc aagaccaacc ggaaggtcac ggttaagcag ctcaaggagg
     2341 actacttcaa gaagattgag tgcttcgatt cggtcgagat ctctggcgtt gaggaccgct
     2401 tcaacgcctc cctggggacc taccacgatc tcctgaagat cattaaggat aaggacttcc
     2461 tggacaacga ggagaatgag gatatcctcg aggacattgt gctgacactc actctgttcg
     2521 aggaccggga gatgatcgag gagcgcctga agacttacgc ccatctcttc gatgacaagg
     2581 tcatgaagca gctcaagagg aggaggtaca ccggctgggg gaggctgagc aggaagctca
     2641 tcaacggcat tcgggacaag cagtccggga agacgatcct cgacttcctg aagagcgatg
     2701 gcttcgcgaa ccgcaatttc atgcagctga ttcacgatga cagcctcaca ttcaaggagg
     2761 atatccagaa ggctcaggtg agcggccagg gggactcgct gcacgagcat atcgcgaacc
     2821 tcgctggctc gccagctatc aagaagggga ttctgcagac cgtgaaggtt gtggacgagc
     2881 tggtgaaggt catgggcagg cacaagcctg agaacatcgt cattgagatg gcccgggaga
     2941 atcagaccac gcagaagggc cagaagaact cacgcgagag gatgaagagg atcgaggagg
     3001 gcattaagga gctggggtcc cagatcctca aggagcaccc ggtggagaac acgcagctgc
     3061 agaatgagaa gctctacctg tactacctcc agaatggccg cgatatgtat gtggaccagg
     3121 agctggatat taacaggctc agcgattacg acgtcgatgc catcgttcca cagtcattcc
     3181 tgaaggatga ctccattgac aacaaggtcc tcaccaggtc ggacaagaac cggggcaagt
     3241 ctgataatgt tccttcagag gaggtcgtta agaagatgaa gaactactgg cgccagctcc
     3301 tgaatgccaa gctgatcacg cagcggaagt tcgataacct cacaaaggct gagaggggcg
     3361 ggctctctga gctggacaag gcgggcttca tcaagaggca gctggtcgag acacggcaga
     3421 tcactaagca cgttgcgcag attctcgact cacggatgaa cactaagtac gatgagaatg
     3481 acaagctgat ccgcgaggtg aaggtcatca ccctgaagtc aaagctcgtc tccgacttca
     3541 ggaaggattt ccagttctac aaggttcggg agatcaacaa ttaccaccat gcccatgacg
     3601 cgtacctgaa cgcggtggtc ggcacagctc tgatcaagaa gtacccaaag ctcgagagcg
     3661 agttcgtgta cggggactac aaggtttacg atgtgaggaa gatgatcgcc aagtcggagc
     3721 aggagattgg caaggctacc gccaagtact tcttctactc taacattatg aatttcttca
     3781 agacagagat cactctggcc aatggcgaga tccggaagcg ccccctcatc gagacgaacg
     3841 gcgagacggg ggagatcgtg tgggacaagg gcagggattt cgcgaccgtc aggaaggttc
     3901 tctccatgcc acaagtgaat atcgtcaaga agacagaggt ccagactggc gggttctcta
     3961 aggagtcaat tctgcctaag cggaacagcg acaagctcat cgcccgcaag aaggactggg
     4021 atccgaagaa gtacggcggg ttcgacagcc ccactgtggc ctactcggtc ctggttgtgg
     4081 cgaaggttga gaagggcaag tccaagaagc tcaagagcgt gaaggagctg ctggggatca
     4141 cgattatgga gcgctccagc ttcgagaaga acccgatcga tttcctggag gcgaagggct
     4201 acaaggaggt gaagaaggac ctgatcatta agctccccaa gtactcactc ttcgagctgg
     4261 agaacggcag gaagcggatg ctggcttccg ctggcgagct gcagaagggg aacgagctgg
     4321 ctctgccgtc caagtatgtg aacttcctct acctggcctc ccactacgag aagctcaagg
     4381 gcagccccga ggacaacgag cagaagcagc tgttcgtcga gcagcacaag cattacctcg
     4441 acgagatcat tgagcagatt tccgagttct ccaagcgcgt gatcctggcc gacgcgaatc
     4501 tggataaggt cctctccgcg tacaacaagc accgcgacaa gccaatcagg gagcaggctg
     4561 agaatatcat tcatctcttc accctgacga acctcggcgc ccctgctgct ttcaagtact
     4621 tcgacacaac tatcgatcgc aagaggtaca caagcactaa ggaggtcctg gacgcgaccc
     4681 tcatccacca gtcgattacc ggcctctacg agacgcgcat cgacctgtct cagctcgggg
     4741 gcgacaagcg gccagcggcg acgaagaagg cggggcaggc gaagaagaag aagtccgggt
     4801 cctcggggtc gccaaagaag aagcggaagg tcggctcaag cgggtcacca aagaagaagc
     4861 ggaaggtcgg ctcgatggac gctaagtccc tcaccgcttg gagccgcacc ctggtgacgt
     4921 tcaaggacgt gttcgtcgat ttcacaaggg aggagtggaa gctcctggac actgctcagc
     4981 agatcgttta ccggaacgtg atgctggaga actacaagaa cctcgtgtcc ctcggctacc
     5041 agctgacgaa gccggatgtt attctccgcc tggagaaggg cgaggagccc gggtccccta
     5101 agaagaagcg gaaggtcggc tcctacccat acgatgtccc tgactacgcc tcatgagctc
     5161 aggcctccat acaagtattg gggatccact acgaatttcc ccgatcgttc aaacatttgg
     5221 caataaagtt tcttaagatt gaatcctgtt gccggtcttg cgatgattat catataattt
     5281 ctgttgaatt acgttaagca tgtaataatt aacatgtaat gcatgacgtt atttatgaga
     5341 tgggttttta tgattagagt cccgcaatta tacatttaat acgcgataga aaacaaaata
     5401 tagcgcgcaa actaggataa attatcgcgc gcggtgtcat ctatgttact agatcgggaa
     5461 ttcgaggcgg tttgcgtatt ggctagagca gcttgccaac atggtggagc acgacactct
     5521 cgtctactcc aagaatatca aagatacagt ctcagaagac caaagggcta ttgagacttt
     5581 tcaacaaagg gtaatatcgg gaaacctcct cggattccat tgcccagcta tctgtcactt
     5641 catcaaaagg acagtagaaa aggaaggtgg cacctacaaa tgccatcatt gcgataaagg
     5701 aaaggctatc gttcaagatg cctctgccga cagtggtccc aaagatggac ccccacccac
     5761 gaggagcatc gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag tggattgatg
     5821 tgaacatggt ggagcacgac actctcgtct actccaagaa tatcaaagat acagtctcag
     5881 aagaccaaag ggctattgag acttttcaac aaagggtaat atcgggaaac ctcctcggat
     5941 tccattgccc agctatctgt cacttcatca aaaggacagt agaaaaggaa ggtggcacct
     6001 acaaatgcca tcattgcgat aaaggaaagg ctatcgttca agatgcctct gccgacagtg
     6061 gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca
     6121 cgtcttcaaa gcaagtggat tgatgtgata tctccactga cgtaagggat gacgcacaat
     6181 cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga
     6241 cacgctgaaa tcaccagtct ctctctacaa atctatctct ctcgagcttt cgcagatccg
     6301 gggggcaatg agatatgaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga
     6361 tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg
     6421 ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg
     6481 gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg
     6541 aagtgcttga cattggggag tttagcgaga gcctgaccta ttgcatctcc cgccgttcac
     6601 agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttcta caaccggtcg
     6661 cggaggctat ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat
     6721 tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg
     6781 atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc
     6841 aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc
     6901 acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg
     6961 actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga
     7021 ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc
     7081 ttgcaggatc gccacgactc cgggcgtata tgctccgcat tggtcttgac caactctatc
     7141 agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa
     7201 tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg
     7261 tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc
     7321 gtccgagggc aaagaaatag agtagatgcc gaccgggatc tgtcgatcga caagctcgag
     7381 tttctccata ataatgtgtg agtagttccc agataaggga attagggttc ctatagggtt
     7441 tcgctcatgt gttgagcata taagaaaccc ttagtatgta tttgtatttg taaaatactt
     7501 ctatcaataa aatttctaat tcctaaaacc aaaatccagt actaaaatcc agatcccccg
     7561 aattaattcg gcgttaattc agtacattaa aaacgtccgc aatgtgttat taagttgtca
     7621 ctagtcaggt taactcaatt cggcgttaat tcagtacatt aaaaacgtcc gcaatgtgtt
     7681 attaagttgt ctaagcgtca atttgtttac accacaatat atcctgccac cagccagcca
     7741 acagctcccc gaccggcagc tcggcacaaa atcaccactc gatacaggca gcccatcagt
     7801 ccgggacggc gtcagcggga gagccgttgt aaggcggcag actttgctca tgttaccgat
     7861 gctattcgga agaacggcaa ctaagctgcc gggtttgaaa cacggatgat ctcgcggagg
     7921 gtagcatgtt gattgtaacg atgacagagc gttgctgcct gtgatcactt aagtaactaa
     7981 ctaacaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
     8041 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
     8101 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
     8161 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
     8221 gcagttccct actctcgctt agtagttaga cgtccccgag atccatgcta gaccatgaat
     8281 ccagaagccc gagaggttgc cgcctttcgg gctttttctt tttcaaaaaa aaaaatttat
     8341 aaaacgatct gttgcggccg gccgccgggt tgtgggcaaa ggcgctcgac ggtgggcaac
     8401 cgcttgcggt tgtccacggg cggagccggt gcgcgtagcg cattgtccac aagccaaggg
     8461 cgaccaataa ttgatatata tattcataat tgaaaagcta attgaacata ctacttgctg
     8521 taactacttg ccggagcgag gggtgtttgc aagctgttga tctgaaaggg ctattagcgt
     8581 tctcacgtgc ctttttgatt agcgatttca cgtgacctta ttagcgattt cacgtactcc
     8641 gattagcgat ttcacgtacc ctgattagcg atttcacgtg gatagttttt ggagcgggcc
     8701 ggaaagcccc gtgaatcaag gctttgcggg gcattagcgg tttcacgtgg ataactaccc
     8761 tctatccaca ggcttccggg gataaaaaag cccgctcgac ggcgggctgt tggatgggga
     8821 tctagcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     8881 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     8941 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     9001 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     9061 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     9121 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     9181 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     9241 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     9301 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     9361 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     9421 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     9481 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     9541 tacggggtct gacgctcagt ggaacggggc ccaatctgaa taatgttaca accaattaac
     9601 caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     9661 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     9721 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     9781 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     9841 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
     9901 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     9961 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
    10021 aggaatcgaa tgcaaccggc gcagggacac tgccagcgca tcaacaatat tttcacctga
    10081 atcaggatat tcttctaata cctggaatgc tgtttttccg gggatcgcag tggtgagtaa
    10141 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
    10201 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
    10261 tttcagaaac aactctggcg catcgggctt cccatacaag cgatagattg tcgcacctga
    10321 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
    10381 taatcgcggc ctcgacgttt cccgttgaat atggctcata acaccccttg tattactgtt
    10441 tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca
    10501 tcagagattt tgagacacgg gccagagctg cagtttgatc ccgaggggaa ccctgtggtt
    10561 gacatgcaca tacaaatgga cgaacggata aaccttttca cgccctttta aatatccgtt
    10621 attctaataa acgctctttt ctcttaggtt tacccgccaa tatatcctgt caaacactga
    10681 tagtttaaac tgaaggcggg aaacgacaat ctgatccaag ctcaagctaa gctcacgtga
    10741 cggaattaag cttgcatgcc tgcaggtcaa catggtggag cacgacacac ttgtctactc
    10801 caaaaatatc aaagatacag tctcagaaga ccaaagggca attgagactt ttcaacaaag
    10861 ggtaatatcc ggaaacctcc tcggattcca ttgcccagct atctgtcact ttattgtgaa
    10921 gatagtggaa aaggaaggtg gctcctacaa atgccatcat tgcgataaag gaaaggccat
    10981 cgttgaagat gcctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat
    11041 cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgataacat
    11101 ggtggagcac gacacacttg tctactccaa aaatatcaaa gatacagtct cagaagacca
    11161 aagggcaat
//
