pSLQ1658-dCas9-EGFP

LOCUS       pSLQ1658-dCas9-EGFP               11177 bp ds-DNA     circular SYN 10-JAN-2018
DEFINITION  Template for NLS-dCas9-NLS-EGFP fusion protein for CRISPR imaging 
            (the recipient vector can be TetON 3G promoter system).
ACCESSION   .
VERSION     .
KEYWORDS    pSLQ1658-dCas9-EGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11177)
  AUTHORS   Chen B, Gilbert LA, Cimini BA, Schnitzbauer J, Zhang W, Li GW, Park 
            J, Blackburn EH, Weissman JS, Qi LS, Huang B
  TITLE     Dynamic Imaging of Genomic Loci in Living Human Cells by an 
            Optimized CRISPR/Cas System.
  JOURNAL   Cell. 2013 Dec 19;155(7):1479-91. doi: 10.1016/j.cell.2013.12.001.
  PUBMED    24360272
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|biofeng.com|
COMMENT     VNTNAME|pSLQ1658-dCas9-EGFP|
FEATURES             Location/Qualifiers
     source          1..11177
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1417..1426
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1432..1452
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1453..5553
                     /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"
     CDS             5563..5583
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5593..6309
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5635..5656)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5896..5915)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6243..6264
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        6320..6819
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6355..6378)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6752..6771
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6840..7439
                     /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(6840..6859)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7336..7356
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             7539..8053
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(8222..8238)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8222..8238)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8235..8257)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8246..8262
                     /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)."
     promoter        complement(8270..8300)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8315..8336
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8453..8470)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8624..9212)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8704..8723)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9383..10243)
                     /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     10006..10025
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10244..10348)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     10416..10434
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(10472..10494)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     10594..10613
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10807..10829
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10957..10976
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             11176..515
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatctcgcca ccatggctag ccccaaaaag
     1441 aagaggaaag tggacaagaa gtattctatc ggactggcca tcgggactaa tagcgtcggg
     1501 tgggccgtga tcactgacga gtacaaggtg ccctctaaga agttcaaggt gctcgggaac
     1561 accgaccggc attccatcaa gaaaaatctg atcggagctc tcctctttga ttcaggggag
     1621 accgctgaag caacccgcct caagcggact gctagacggc ggtacaccag gaggaagaac
     1681 cggatttgtt accttcaaga gatattctcc aacgaaatgg caaaggtcga cgacagcttc
     1741 ttccataggc tggaagaatc attcctcgtg gaagaggata agaagcatga acggcatccc
     1801 atcttcggta atatcgtcga cgaggtggcc tatcacgaga aatacccaac catctaccat
     1861 cttcgcaaaa agctggtgga ctcaaccgac aaggcagacc tccggcttat ctacctggcc
     1921 ctggcccaca tgatcaagtt cagaggccac ttcctgatcg agggcgacct caatcctgac
     1981 aatagcgatg tggataaact gttcatccag ctggtgcaga cttacaacca gctctttgaa
     2041 gagaacccca tcaatgcaag cggagtcgat gccaaggcca ttctgtcagc ccggctgtca
     2101 aagagccgca gacttgagaa tcttatcgct cagctgccgg gtgaaaagaa aaatggactg
     2161 ttcgggaacc tgattgctct ttcacttggg ctgactccca atttcaagtc taatttcgac
     2221 ctggcagagg atgccaagct gcaactgtcc aaggacacct atgatgacga tctcgacaac
     2281 ctcctggccc agatcggtga ccaatacgcc gaccttttcc ttgctgctaa gaatctttct
     2341 gacgccatcc tgctgtctga cattctccgc gtgaacactg aaatcaccaa ggcccctctt
     2401 tcagcttcaa tgattaagcg gtatgatgag caccaccagg acctgaccct gcttaaggca
     2461 ctcgtccggc agcagcttcc ggagaagtac aaggaaatct tctttgacca gtcaaagaat
     2521 ggatacgccg gctacatcga cggaggtgcc tcccaagagg aattttataa gtttatcaaa
     2581 cctatccttg agaagatgga cggcaccgaa gagctcctcg tgaaactgaa tcgggaggat
     2641 ctgctgcgga agcagcgcac tttcgacaat gggagcattc cccaccagat ccatcttggg
     2701 gagcttcacg ccatccttcg gcgccaagag gacttctacc cctttcttaa ggacaacagg
     2761 gagaagattg agaaaattct cactttccgc atcccctact acgtgggacc cctcgccaga
     2821 ggaaatagcc ggtttgcttg gatgaccaga aagtcagaag aaactatcac tccctggaac
     2881 ttcgaagagg tggtggacaa gggagccagc gctcagtcat tcatcgaacg gatgactaac
     2941 ttcgataaga acctccccaa tgagaaggtc ctgccgaaac attccctgct ctacgagtac
     3001 tttaccgtgt acaacgagct gaccaaggtg aaatatgtca ccgaagggat gaggaagccc
     3061 gcattcctgt caggcgaaca aaagaaggca attgtggacc ttctgttcaa gaccaataga
     3121 aaggtgaccg tgaagcagct gaaggaggac tatttcaaga aaattgaatg cttcgactct
     3181 gtggagatta gcggggtcga agatcggttc aacgcaagcc tgggtaccta ccatgatctg
     3241 cttaagatca tcaaggacaa ggattttctg gacaatgagg agaacgagga catccttgag
     3301 gacattgtcc tgactctcac tctgttcgag gaccgggaaa tgatcgagga gaggcttaag
     3361 acctacgccc atctgttcga cgataaagtg atgaagcaac ttaaacggag aagatatacc
     3421 ggatggggac gccttagccg caaactcatc aacggaatcc gggacaaaca gagcggaaag
     3481 accattcttg atttccttaa gagcgacgga ttcgctaatc gcaacttcat gcaacttatc
     3541 catgatgatt ccctgacctt taaggaggac atccagaagg cccaagtgtc tggacaaggt
     3601 gactcactgc acgagcatat cgcaaatctg gctggttcac ccgctattaa gaagggtatt
     3661 ctccagaccg tgaaagtcgt ggacgagctg gtcaaggtga tgggtcgcca taaaccagag
     3721 aacattgtca tcgagatggc cagggaaaac cagactaccc agaagggaca gaagaacagc
     3781 agggagcgga tgaaaagaat tgaggaaggg attaaggagc tcgggtcaca gatccttaaa
     3841 gagcacccgg tggaaaacac ccagcttcag aatgagaagc tctatctgta ctaccttcaa
     3901 aatggacgcg atatgtatgt ggaccaagag cttgatatca acaggctctc agactacgac
     3961 gtggacgcca tcgtccctca gagcttcctc aaagacgact caattgacaa taaggtgctg
     4021 actcgctcag acaagaaccg gggaaagtca gataacgtgc cctcagagga agtcgtgaaa
     4081 aagatgaaga actattggcg ccagcttctg aacgcaaagc tgatcactca gcggaagttc
     4141 gacaatctca ctaaggctga gaggggcgga ctgagcgaac tggacaaagc aggattcatt
     4201 aaacggcaac ttgtggagac tcggcagatt actaaacatg tcgcccaaat ccttgactca
     4261 cgcatgaata ccaagtacga cgaaaacgac aaacttatcc gcgaggtgaa ggtgattacc
     4321 ctgaagtcca agctggtcag cgatttcaga aaggactttc aattctacaa agtgcgggag
     4381 atcaataact atcatcatgc tcatgacgca tatctgaatg ccgtggtggg aaccgccctg
     4441 atcaagaagt acccaaagct ggaaagcgag ttcgtgtacg gagactacaa ggtctacgac
     4501 gtgcgcaaga tgattgccaa atctgagcag gagatcggaa aggccaccgc aaagtacttc
     4561 ttctacagca acatcatgaa tttcttcaag accgaaatca cccttgcaaa cggtgagatc
     4621 cggaagaggc cgctcatcga gactaatggg gagactggcg aaatcgtgtg ggacaagggc
     4681 agagatttcg ctaccgtgcg caaagtgctt tctatgcctc aagtgaacat cgtgaagaaa
     4741 accgaggtgc aaaccggagg cttttctaag gaatcaatcc tccccaagcg caactccgac
     4801 aagctcattg caaggaagaa ggattgggac cctaagaagt acggcggatt cgattcacca
     4861 actgtggctt attctgtcct ggtcgtggct aaggtggaaa aaggaaagtc taagaagctc
     4921 aagagcgtga aggaactgct gggtatcacc attatggagc gcagctcctt cgagaagaac
     4981 ccaattgact ttctcgaagc caaaggttac aaggaagtca agaaggacct tatcatcaag
     5041 ctcccaaagt atagcctgtt cgaactggag aatgggcgga agcggatgct cgcctccgct
     5101 ggcgaacttc agaagggtaa tgagctggct ctcccctcca agtacgtgaa tttcctctac
     5161 cttgcaagcc attacgagaa gctgaagggg agccccgagg acaacgagca aaagcaactg
     5221 tttgtggagc agcataagca ttatctggac gagatcattg agcagatttc cgagttttct
     5281 aaacgcgtca ttctcgctga tgccaacctc gataaagtcc ttagcgcata caataagcac
     5341 agagacaaac caattcggga gcaggctgag aatatcatcc acctgttcac cctcaccaat
     5401 cttggtgccc ctgccgcatt caagtacttc gacaccacca tcgaccggaa acgctatacc
     5461 tccaccaaag aagtgctgga cgccaccctc atccaccaga gcatcaccgg actttacgaa
     5521 actcggattg acctctcaca gctcggaggg gatgagggag ctcccaagaa aaagcgcaag
     5581 gtaggtagtt ccgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     5641 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     5701 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     5761 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     5821 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     5881 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     5941 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     6001 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     6061 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     6121 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     6181 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     6241 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     6301 tacaagtaag aattctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc
     6361 gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat
     6421 tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct
     6481 tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg
     6541 tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa
     6601 tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg
     6661 gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg
     6721 gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct
     6781 gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagccc aagcttacca
     6841 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     6901 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     6961 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     7021 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     7081 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     7141 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     7201 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     7261 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     7321 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     7381 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac
     7441 gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc acgaccccat gcatcgataa
     7501 aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt
     7561 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat
     7621 agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata
     7681 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
     7741 tcccgccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
     7801 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
     7861 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct
     7921 ccgatagact gcgtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc
     7981 gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc
     8041 gggggtcttt catgggtaac agtttcttga agttggagaa caacattctg agggtaggag
     8101 tcgaatatta agtaatcctg actcaattag ccactgtttt gaatccacat actccaatac
     8161 tcctgaaata gttcattatg gacagcgcag aagagctggg gagaattaat tcgtaatcat
     8221 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     8281 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     8341 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     8401 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     8461 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     8521 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     8581 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     8641 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     8701 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     8761 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     8821 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     8881 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     8941 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     9001 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     9061 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     9121 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     9181 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     9241 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     9301 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     9361 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     9421 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     9481 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     9541 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     9601 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     9661 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     9721 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     9781 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     9841 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     9901 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     9961 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
    10021 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
    10081 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
    10141 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
    10201 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
    10261 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
    10321 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
    10381 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
    10441 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
    10501 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
    10561 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
    10621 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
    10681 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
    10741 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
    10801 ggttttccca gtcacgacgt tgtaaaacga cggcgcaagg aatggtgcat gcaaggagat
    10861 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
    10921 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
    10981 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggcgatt
    11041 agtccaattt gttaaagaca ggatatcagt ggtccaggct ctagttttga ctcaacaata
    11101 tcaccagctg aagcctatag agtacgagcc atagataaaa taaaagattt tatttagtct
    11161 ccagaaaaag gggggaa
//