LOCUS       Exported               10663 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses sgRNA and Cas9-Puro in Drosophila S2 cells.
ACCESSION   .
VERSION     .
KEYWORDS    pAc-sgRNA-Cas9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10663)
  AUTHORS   Bassett AR, Tibbit C, Ponting CP, Liu JL
  TITLE     Mutagenesis and homologous recombination in Drosophila cell lines 
            using CRISPR/Cas9.
  JOURNAL   Biol Open. 2013 Dec 10. pii: bio.20137120v1. doi: 
            10.1242/bio.20137120.
  PUBMED    24326186
REFERENCE   2  (bases 1 to 10663)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10663
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_RNA        complement(13..88)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(107..506)
                     /label=dU6-2 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-2 
                     snRNA (Das et al., 1987)"
     promoter        507..2948
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5c promoter"
     CDS             2992..3057
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3064..3084
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3109..7209
                     /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             7210..7257
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7273..7326
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             7351..7950
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7351..7370)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7847..7867
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7958..8092
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7982..8001)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8036..8055
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(8714..9302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8794..8813)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9473..10333)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     10096..10115
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10334..10438)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     10506..10524
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 agatctaaaa aagcaccgac tcggtgccac tttttcaagt tgataacgga ctagccttat
       61 tttaacttgc tatttctagc tctaaaacgg aagagctcgc tcttccgaag tattgaggaa
      121 aacataccta tataaatgat caacatcagg aaagagcagt tgagaattat aagaattggc
      181 aaatggtctt aagaaccctc tgcttaagat tttcaaaatt tcctttagaa tcaaagtgtc
      241 ctattgttcg tttgttgaaa acacagttcg atttattgac tataataaat tgatagtttt
      301 aaatatagag gcacgactaa gagagtttgg ttttgttttg gttagggacc aaaaaaagta
      361 tacataacga ataaaaaagg atttaagtgc aaatggtaaa aaagtgtcga gtttttcttg
      421 agttgattgt gctgtaatga gactctgcat tcggcatttg actcggcttt tcctactcgt
      481 gccgtatttc aggctgcaag tcgaacagat ctcgctgcct gttatgtggc ccacaaacca
      541 agacacgttt tatggccatt aaagctggct gatcgtcgcc aaacaccaaa tacataatga
      601 atatgtacac attcgagaaa gaagcgatca aagaagcgtc ttcgggcgga gtaggagaat
      661 gcggaggaga aggagaacga gctgatctag tatctctcca caatccaatg ccaactgacc
      721 aactggccat attcggagca atttgaagcc aatttccatc gcctggcgat cgctccattc
      781 ttggctatat gtttttcacc gttacccggg gccattttca aagactcgtc ggcaagataa
      841 gattgtgtca ctcgctgtct ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga
      901 cgtcggcgag tattttgaag aatgagaata atttgtattt atacgaaaat cagttagtgg
      961 aattttctac aaaaacatgt tatctataga taattttgtt gcaaaatatg ttgactatga
     1021 caaagattgt atgtatatac ctttaatgta ttctcatttt cttatgtatt tataatggca
     1081 atgatgatac tgatgatatt ttaagatgat gccagaccac aggctgattt ctgcgtcttt
     1141 tgccgaacgc agtgcatgtg cggttgttgt tttttggaat agtttcaatt ttcggactgt
     1201 ccgctttgat ttcagtttct tggcttattc aaaaagcaaa gtaaagccaa aaaagcgaga
     1261 tggcaatacc aaatgcggca aaacggtagt ggaaggaaag gggtgcgggg cagcggaagg
     1321 aagggtgggg cggggcgtgg cggggtctgt ggctgggcgc gacgtcaccg acgttggagc
     1381 cactcctttg accatgtgtg cgtgtgtgta ttattcgtgt ctcgccactc gccggttgtt
     1441 tttttctttt tatctcgctc tctctagcgc catctcgtac gcatgctcaa cgcaccgcat
     1501 gttgccgtgt cctttatgcg tcattttggc tcgaaatagg caattattta aacaaagatt
     1561 agtcaacgaa aacgctaaaa taaataagtc tacaatatgg ttacttattg ccatgtgtgt
     1621 gcagccaacg atagcaacaa aagcaacaac acagtggctt tccctctttc actttttgtt
     1681 tgcaagcgcg tgcgagcaag acggcacgac cggcaaacgc aattacgctg acaaagagca
     1741 gacgaagttt tggccgaaaa acatcaaggc gcctgatacg aatgcatttg caataacaat
     1801 tgcgatattt aatattgttt atgaagctgt ttgacttcaa aacacacaaa aaaaaaaata
     1861 aaacaaatta tttgaaagag aattaggaat cggacagctt atcgttacgg gctaacagca
     1921 caccgagacg aaatagctta cctgacgtca cagcctctgg aagaactgcc gccaagcaga
     1981 gagagagaga aaaagaggga gagcagctta gaccgcatgt gcttgtgtgt gaggcgtctc
     2041 tctcttcgtc tcctgtttgc gcaaacgcat agactgcact gagaaaatcg attacctatt
     2101 ttttatgaat gaatatttgc actattacta ttcaaaacta ttaagatagc aatcacattc
     2161 aatagccaaa tactatacca cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg
     2221 ctgcatattt aggacggcat cattatagaa atgcttcttg ctgtgtactt ttctctcgtc
     2281 tggcagctgt ttcgccgtta ttgttaaaac cggcttaagt taggtgtgtt ttctacgact
     2341 agtgatgccc ctactagaag atgtgtgttg cacaaatgtc cctgaataac caatttgaag
     2401 tgcagatagc agtaaacgta agctaatatg aatattattt aactgtaatg ttttaatatc
     2461 gctggacatt actaataaac ccactataaa cacatgtaca tatgtatgtt ttggcataca
     2521 atgagtagtt ggggaaaaaa tgtgtaaaag caccgtgacc atcacagcat aaagataacc
     2581 agctgaagta tcgaatatga gtaaccccca aattgaatca catgccgcaa ctgataggac
     2641 ccatggaagt acactcttca tggcgatata caagacacac acaagcacga acacccagtt
     2701 gcggaggaaa ttctccgtaa atgaaaaccc aatcggcgaa caattcatac ccatatatgg
     2761 taaaagtttt gaacgcgact tgagagcgga gagcattgcg gctgataagg ttttagcgct
     2821 aagcgggctt tataaaacgg gctgcgggac cagttttcat atcactaccg tttgagttct
     2881 tgtgctgtgt ggatactcct cccgacacaa agccgctcca tcagccagca gtcgtctaat
     2941 ccagagaccc cggatcgggg tacctactag tccagtgtgg tggaattcat ggactataag
     3001 gaccacgacg gagactacaa ggatcatgat attgattaca aagacgatga cgataagatg
     3061 gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccga caagaagtac
     3121 agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac
     3181 aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag
     3241 aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag
     3301 agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc
     3361 ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc
     3421 ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag
     3481 gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc
     3541 accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg
     3601 ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc
     3661 atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc
     3721 gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg
     3781 atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc
     3841 ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag
     3901 ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag
     3961 tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc
     4021 ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac
     4081 gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag
     4141 aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc
     4201 ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc
     4261 accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc
     4321 gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg
     4381 caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc
     4441 ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg
     4501 accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc
     4561 gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag
     4621 aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc
     4681 aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa
     4741 aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa
     4801 gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat
     4861 cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac
     4921 ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg
     4981 tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac
     5041 aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag
     5101 ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc
     5161 gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa
     5221 gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc
     5281 aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac
     5341 gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga
     5401 gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa
     5461 gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag
     5521 ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac
     5581 caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc
     5641 tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc
     5701 aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag
     5761 ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga
     5821 ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg
     5881 cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag
     5941 aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat
     6001 ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac
     6061 gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa
     6121 agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc
     6181 gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt
     6241 ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca
     6301 aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa
     6361 gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc
     6421 agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac
     6481 tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg
     6541 gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg
     6601 atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag
     6661 ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag
     6721 ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa
     6781 ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg
     6841 aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac
     6901 ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct
     6961 aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag
     7021 gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag
     7081 tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc
     7141 accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg
     7201 ggaggcgaca aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa gaaaaaggga
     7261 tcaggcaagc ttgagggcag aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc
     7321 ggccctgcta gcggtagcgg cagcggtagc atgaccgagt acaagcccac ggtgcgcctc
     7381 gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac
     7441 cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac cgagctgcaa
     7501 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     7561 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     7621 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     7681 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     7741 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     7801 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     7861 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     7921 tggtgcatga cccgcaagcc cggtgcctga ctcgagggat ccagacatga taagatacat
     7981 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     8041 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     8101 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     8161 gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgaccgatg cccttgagag
     8221 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
     8281 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt
     8341 tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg
     8401 gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg
     8461 agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt
     8521 tcgcgacgcg aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg
     8581 ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc
     8641 ttcaaggatc gctcgcggct cttaccagcc agcaaaaggc caggaaccgt aaaaaggccg
     8701 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     8761 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     8821 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     8881 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     8941 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     9001 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     9061 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     9121 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     9181 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     9241 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     9301 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     9361 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     9421 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     9481 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     9541 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     9601 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     9661 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     9721 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     9781 ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     9841 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     9901 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     9961 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
    10021 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
    10081 cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
    10141 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
    10201 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
    10261 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
    10321 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
    10381 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
    10441 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
    10501 ataaaaatag gcgtatcacg aggccctttc gtcttcaagg aattaattct atattctaaa
    10561 aacacaaatg atacttctaa aaaaaatcat gaatggcatc aactctgaat caaatctttg
    10621 cagatgcacc tacttctcat ttccactgtc acatcatttt tcc
//
