pSpCas9(BB)-2A-Blast

LOCUS        pSpCas9(BB)-2A-Blast               8967 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Used for expression of sgRNA in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pSpCas9(BB)-2A-Blast
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8967)
  TITLE     pSpCas9(BB)-2A-Blast
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8967)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8967
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        651..891
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     651..671
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     822..841
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        918..993
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        1090..1375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1377..1654
                     /label=chicken beta-actin promoter
     intron          1655..1882
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1894..1903
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1903..1968
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1975..1995
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2020..6120
                     /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             6121..6168
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6178..6234
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             6235..6630
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(6658..6675)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6664..6871
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6880..7020
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6880..7009
                     /label=AAV2 ITR
     rep_origin      7095..7550
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7182..7201)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7392..7413
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7567..7586)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7686..7708
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7746..7764)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7832..7936
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7937..8797
                     /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(8155..8174)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt gagggcctat
      661 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa
      721 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
      781 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
      841 taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg
      901 ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
      961 tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct agaaatagca
     1021 agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac aaatggctct
     1081 agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     1141 ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc aatgggtgga
     1201 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     1261 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt acatgacctt
     1321 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga
     1381 ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt
     1441 gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggggcg
     1501 cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc
     1561 ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg
     1621 gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc
     1681 cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact
     1741 cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc tgagcaagag
     1801 gtaagggttt aagggatggt tggttggtgg ggtattaatg tttaattacc tggagcacct
     1861 gcctgaaatc actttttttc aggttggacc ggtgccacca tggactataa ggaccacgac
     1921 ggagactaca aggatcatga tattgattac aaagacgatg acgataagat ggccccaaag
     1981 aagaagcgga aggtcggtat ccacggagtc ccagcagccg acaagaagta cagcatcggc
     2041 ctggacatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc
     2101 agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc
     2161 ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc
     2221 agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac
     2281 gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa
     2341 gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac
     2401 cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag
     2461 gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc
     2521 ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg
     2581 gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc
     2641 aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag
     2701 ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg
     2761 acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag
     2821 gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac
     2881 ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg
     2941 aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac
     3001 caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa
     3061 gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc
     3121 caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa
     3181 ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc
     3241 agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat
     3301 ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc
     3361 ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag
     3421 agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc
     3481 cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg
     3541 cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa
     3601 tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc
     3661 gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac
     3721 ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac
     3781 gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac
     3841 aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac
     3901 agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg
     3961 aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac
     4021 ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc
     4081 gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc
     4141 cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc
     4201 ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg
     4261 aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag
     4321 accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc
     4381 aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac
     4441 gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg
     4501 gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag
     4561 gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac
     4621 aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac
     4681 gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg
     4741 agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca
     4801 aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag
     4861 ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag
     4921 gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac
     4981 ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc
     5041 gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa
     5101 atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc
     5161 gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa
     5221 accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc
     5281 atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag
     5341 tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct
     5401 aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa
     5461 gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc
     5521 atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa
     5581 gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac
     5641 ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg
     5701 ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc
     5761 cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag
     5821 atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac
     5881 aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat
     5941 atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac
     6001 accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc
     6061 caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac
     6121 aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa agaaaaagga attcggcgca
     6181 acaaacttct ctctgctgaa acaagccgga gatgtcgaag agaatcctgg accggccaag
     6241 cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc
     6301 cccatctctg aggactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc
     6361 actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc
     6421 actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac
     6481 aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg
     6541 atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg
     6601 ctgccctctg gttatgtgtg ggagggctaa gaattctaac tagagctcgc tgatcagcct
     6661 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     6721 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     6781 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     6841 attgggaaga gaatagcagg catgctgggg agcggccgca ggaaccccta gtgatggagt
     6901 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     6961 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     7021 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     7081 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     7141 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     7201 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     7261 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     7321 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     7381 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     7441 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     7501 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     7561 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     7621 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     7681 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     7741 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     7801 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     7861 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     7921 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     7981 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     8041 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     8101 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     8161 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     8221 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     8281 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     8341 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     8401 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     8461 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     8521 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     8581 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     8641 cggtgagcgt ggaagccgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     8701 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     8761 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     8821 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     8881 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     8941 ccccgtagaa aagatcaaag gatcttc
//