pEF1a-nCas9-BFP

LOCUS       pEF1a-nCas9-BFP          8970 bp DNA     circular SYN 07-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8970)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 20, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     77..142
                     /label=OL2624
     primer_bind     80..122
                     /label=OL2675
     promoter        106..1284
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     106..122
                     /label=OL2712
     intron          337..1275
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     359..376
                     /label=ef1a-seq1
     primer_bind     617..630
                     /label=OL1324
     primer_bind     1135..1154
                     /label=ef1a-seq2
     primer_bind     1232..1252
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(1256..1309)
                     /label=OL2676
     primer_bind     complement(1285..1313)
                     /label=OL2625
     primer_bind     1291..1310
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1291..1309
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1291..1309
                     /label=OL1124
     primer_bind     complement(1306..1325)
                     /label=OL1206
     primer_bind     complement(1353..1379)
                     /label=OL1013
     primer_bind     complement(1357..1379)
                     /label=OL1036
     CDS             1359..1379
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     1360..1401
                     /label=OL1039
     primer_bind     1360..1399
                     /label=OL1296
     primer_bind     complement(1360..1379)
                     /label=OL1298
     CDS             1380..5480
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(R221K N394K H840A)
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRKLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLKREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(1384..1403)
                     /label=OL1051
     primer_bind     complement(1916..1935)
                     /label=OL989
     primer_bind     complement(1988..1998)
                     /label=OL900
     primer_bind     complement(2021..2050)
                     /label=OL1321
     primer_bind     2026..2055
                     /label=OL1322
     primer_bind     2188..2202
                     /label=OL1270
     primer_bind     complement(2541..2575)
                     /label=OL1323
     primer_bind     2541..2575
                     /label=OL1324
     primer_bind     3100..3114
                     /label=OL1271
     primer_bind     3846..3860
                     /label=OL1272
     primer_bind     complement(3879..3910)
                     /label=OL1316
     primer_bind     3879..3910
                     /label=OL1317
     primer_bind     5183..5202
                     /label=OL1328
     CDS             5481..5558
                     /label=SGGSx2-sv40NLS-SGGSx2
                     /note="/created_by=chenp"
     primer_bind     5481..5503
                     /label=OL1326
     primer_bind     5520..5536
                     /label=OL1153
     primer_bind     complement(5528..5544)
                     /label=OL1154
     primer_bind     complement(5532..5558)
                     /label=OL1015
     primer_bind     5537..5558
                     /label=OL1014
     primer_bind     5538..5582
                     /label=OL1313
     CDS             5538..5558
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5543..5558)
                     /label=OL1040
     CDS             5559..5615
                     /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"
     primer_bind     5559..5615
                     /label=OL866
     primer_bind     complement(5559..5615)
                     /label=OL867
     primer_bind     complement(5559..5589)
                     /label=OL1019
     primer_bind     5596..5641
                     /label=OL868
     CDS             5616..6311
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     primer_bind     complement(6289..6314)
                     /label=OL1018
     primer_bind     complement(6290..6335)
                     /label=OL1314
     primer_bind     complement(6290..6314)
                     /label=OL2711
     primer_bind     6315..6344
                     /label=OL1014
     primer_bind     complement(6315..6344)
                     /label=OL900
     primer_bind     complement(6315..6341)
                     /label=OL1015
     primer_bind     complement(6315..6339)
                     /label=OL1040
     primer_bind     6315..6334
                     /label=OL1207
     primer_bind     6320..6343
                     /label=OL1035
     CDS             6320..6337
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     6320..6337
                     /label=OL1133
     primer_bind     complement(6320..6337)
                     /label=OL1152
     primer_bind     complement(6360..6377)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6366..6590
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal
                     牛生长激素聚腺苷酸化信号
                     牛生长激素（BGH）多聚腺苷酸化信号能够保证高效的转录终止和mRNA的poly A结构"
     primer_bind     complement(6410..6427)
                     /label=OL698
     primer_bind     complement(6661..6677)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6661..6677)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6674..6696)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6685..6701
                     /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(6709..6739)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6754..6775
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6892..6909)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7063..7651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7143..7162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(7245..7264)
                     /label=OL1041
     primer_bind     complement(7378..7397)
                     /label=OL631
     primer_bind     7434..7453
                     /label=OL798
     primer_bind     7695..7714
                     /label=OL659
     CDS             complement(7822..8682)
                     /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     8445..8464
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     8598..8617
                     /label=OL1033
     primer_bind     complement(8598..8617)
                     /label=OL1034
     promoter        complement(8683..8787)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8866..8885)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
       61 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagggctc cggtgcccgt
      121 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
      181 tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
      241 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
      301 gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc
      361 gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg
      421 cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct
      481 tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc
      541 cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta
      601 gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta
      661 aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg
      721 gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga
      781 atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg
      841 tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa
      901 agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga
      961 gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct
     1021 tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt
     1081 tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact
     1141 gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt
     1201 gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt
     1261 tttcttccat ttcaggtgtc gtgaggccgc taatacgact cactataggg agagccgcca
     1321 ccatgaaacg gacagccgac ggaagcgagt tcgagtcacc aaagaagaag cggaaagtcg
     1381 acaagaagta cagcatcggc ctggacatcg gcaccaactc tgtgggctgg gccgtgatca
     1441 ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca
     1501 gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca
     1561 cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc
     1621 tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg
     1681 aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca
     1741 tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac
     1801 tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga
     1861 tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg
     1921 acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca
     1981 acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagaaagc
     2041 tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggaaacctga
     2101 ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg
     2161 ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga
     2221 tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc
     2281 tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga
     2341 tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc
     2401 agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct
     2461 acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa
     2521 agatggacgg caccgaggaa ctgctcgtga agctgaagag agaggacctg ctgcggaagc
     2581 agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca
     2641 ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga
     2701 agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat
     2761 tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg
     2821 tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc
     2881 tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata
     2941 acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg
     3001 gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga
     3061 agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg
     3121 gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca
     3181 aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga
     3241 ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc
     3301 tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc
     3361 tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt
     3421 tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc
     3481 tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg
     3541 agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga
     3601 aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg
     3661 aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga
     3721 agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg
     3781 aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata
     3841 tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gacgctatcg
     3901 tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca
     3961 agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact
     4021 actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca
     4081 aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg
     4141 tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta
     4201 agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc
     4261 tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc
     4321 accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc
     4381 ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga
     4441 tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca
     4501 tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc
     4561 tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca
     4621 ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga
     4681 caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca
     4741 gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt
     4801 ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag
     4861 agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc
     4921 tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact
     4981 ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga
     5041 agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact
     5101 atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc
     5161 acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc
     5221 tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca
     5281 tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg
     5341 ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg
     5401 tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc
     5461 tgtctcagct gggaggtgac tccggcggaa gctctggtgg cagcaagcgg accgccgacg
     5521 gctctgaatt cgagagccct aagaagaaaa gaaaggtggc tactaacttc agcctgctga
     5581 agcaggctgg agacgtggag gagaaccctg gacctagcga gctgattaag gagaacatgc
     5641 acatgaagct gtacatggag ggcaccgtgg acaaccatca cttcaagtgc acatccgagg
     5701 gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggtggtc gagggcggcc
     5761 ctctcccctt cgccttcgac atcctggcta ctagcttcct ctacggcagc aagaccttca
     5821 tcaaccacac ccagggcatc cccgacttct tcaagcagtc cttccctgag ggcttcacat
     5881 gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc
     5941 tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc acatccaacg
     6001 gccctgtgat gcagaagaaa acactcggct gggaggcctt caccgagacg ctgtaccccg
     6061 ctgacggcgg cctggaaggc agaaacgaca tggccctgaa gctcgtgggc gggagccatc
     6121 tgatcgcaaa catcaagacc acatatagat ccaagaaacc cgctaagaac ctcaagatgc
     6181 ctggcgtcta ctatgtggac tacagactgg aaagaatcaa ggaggccaac aacgagacct
     6241 acgtcgagca gcacgaggtg gcagtggcca gatactgcga cctccctagc aaactggggc
     6301 acaagcttaa ttagccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     6361 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     6421 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgagaaaa ttgcatcgca
     6481 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     6541 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     6601 ggaaagaacc agctggggct cgataccgtc gacctctagc tagagcttgg cgtaatcatg
     6661 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6721 cggaagcata aagtgtaaag cctagggtgc ctaatgagtg agctaactca cattaattgc
     6781 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6841 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6901 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6961 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7021 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7081 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7141 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7201 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7261 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7321 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7381 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7441 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7501 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7561 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7621 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7681 tgacactcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7741 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7801 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7861 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7921 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7981 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8041 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8101 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8161 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8221 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8281 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8341 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8401 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8461 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8521 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8581 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8641 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8701 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8761 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     8821 cggatcggga gatcgatctc ccgatcccct agggtctact ctcagtacaa tctgctctga
     8881 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     8941 cgcgagcaaa atttaagcta caacaaggca
//