LOCUS       sequence-120194-        8211 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of Sp-dCas9 in mammalian cells, under an 
            EF1-alpha promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8211)
  AUTHORS   Shechner DM, Hacisuleyman E, Younger ST, Rinn JL
  TITLE     Multiplexable, locus-specific targeting of long RNAs with 
            CRISPR-Display.
  JOURNAL   Nat Methods. 2015 Jul;12(7):664-70. doi: 10.1038/nmeth.3433. Epub 
            2015 Jun 1.
  PUBMED    26030444
REFERENCE   2  (bases 1 to 8211)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nmeth.3433"; journalName: "Nat Methods"; date: "2015-07"; 
            volume: "12"; issue: "7"; pages: "664-70"
COMMENT     Sequence Label: pEF_dCas9
FEATURES             Location/Qualifiers
     source          1..8211
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(1..792,793..861))
                     /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     624..643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(862..966)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1045..1064)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1261..1472
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          1473..2415
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2372..2392
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      2448..2457
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2463..2483
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2484..5882,5883..5885,5886..6482,6483,6484..6486,
                     6487..6488,6489..6491,6492..6584)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /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             join(2484..2507,2508..2510,2511..4997,4998..5000,
                     5001..6584)
                     /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             6591..6611
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6618..6683
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6710..6727
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6750..6767)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6756..6980
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7051..7067)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7051..7067)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7064..7086)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7075..7091
                     /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(join(7099..7105,7106..7123,7124..7129))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7144..7165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7282..7299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7453..8041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7533..7552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
       61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
      181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
      901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      961 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcgatctcc
     1021 cgatccccta gggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     1081 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     1141 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     1201 gctgcttcgc gatgtacggg ccagatatac gcgttgcgtg aggctccggt gcccgtcagt
     1261 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     1321 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     1381 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     1441 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
     1501 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacg cccctggctg
     1561 cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct
     1621 tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc
     1681 cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta
     1741 gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta
     1801 aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg
     1861 gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga
     1921 atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg
     1981 tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa
     2041 agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga
     2101 gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct
     2161 tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt
     2221 tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact
     2281 gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt
     2341 gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt
     2401 tttcttccat ttcaggtgtc cgctagagat ccgcggccgc gagagccgcc accatggcta
     2461 gccccaaaaa gaagaggaaa gtggataaaa agtattctat tggtttagcc atcggcacta
     2521 attccgttgg atgggctgtc ataaccgatg aatacaaagt accttcaaag aaatttaagg
     2581 tgttggggaa cacagaccgt cattcgatta aaaagaatct tatcggtgcc ctcctattcg
     2641 atagtggcga aacggcagag gcgactcgcc tgaaacgaac cgctcggaga aggtatacac
     2701 gtcgcaagaa ccgaatatgt tacttacaag aaatttttag caatgagatg gccaaagttg
     2761 acgattcttt ctttcaccgt ttggaagagt ccttccttgt cgaagaggac aagaaacatg
     2821 aacggcaccc catctttgga aacatagtag atgaggtggc atatcatgaa aagtacccaa
     2881 cgatttatca cctcagaaaa aagctagttg actcaactga taaagcggac ctgaggttaa
     2941 tctacttggc tcttgcccat atgataaagt tccgtgggca ctttctcatt gagggtgatc
     3001 taaatccgga caactcggat gtcgacaaac tgttcatcca gttagtacaa acctataatc
     3061 agttgtttga agagaaccct ataaatgcaa gtggcgtgga tgcgaaggct attcttagcg
     3121 cccgcctctc taaatcccga cggctagaaa acctgatcgc acaattaccc ggagagaaga
     3181 aaaatgggtt gttcggtaac cttatagcgc tctcactagg cctgacacca aattttaagt
     3241 cgaacttcga cttagctgaa gatgccaaat tgcagcttag taaggacacg tacgatgacg
     3301 atctcgacaa tctactggca caaattggag atcagtatgc ggacttattt ttggctgcca
     3361 aaaaccttag cgatgcaatc ctcctatctg acatactgag agttaatact gagattacca
     3421 aggcgccgtt atccgcttca atgatcaaaa ggtacgatga acatcaccaa gacttgacac
     3481 ttctcaaggc cctagtccgt cagcaactgc ctgagaaata taaggaaata ttctttgatc
     3541 agtcgaaaaa cgggtacgca ggttatattg acggcggagc gagtcaagag gaattctaca
     3601 agtttatcaa acccatatta gagaagatgg atgggacgga agagttgctt gtaaaactca
     3661 atcgcgaaga tctactgcga aagcagcgga ctttcgacaa cggtagcatt ccacatcaaa
     3721 tccacttagg cgaattgcat gctatactta gaaggcagga ggatttttat ccgttcctca
     3781 aagacaatcg tgaaaagatt gagaaaatcc taacctttcg cataccttac tatgtgggac
     3841 ccctggcccg agggaactct cggttcgcat ggatgacaag aaagtccgaa gaaacgatta
     3901 ctccatggaa ttttgaggaa gttgtcgata aaggtgcgtc agctcaatcg ttcatcgaga
     3961 ggatgaccaa ctttgacaag aatttaccga acgaaaaagt attgcctaag cacagtttac
     4021 tttacgagta tttcacagtg tacaatgaac tcacgaaagt taagtatgtc actgagggca
     4081 tgcgtaaacc cgcctttcta agcggagaac agaagaaagc aatagtagat ctgttattca
     4141 agaccaaccg caaagtgaca gttaagcaat tgaaagagga ctactttaag aaaattgaat
     4201 gcttcgattc tgtcgagatc tccggggtag aagatcgatt taatgcgtca cttggtacgt
     4261 atcatgacct cctaaagata attaaagata aggacttcct ggataacgaa gagaatgaag
     4321 atatcttaga agatatagtg ttgactctta ccctctttga agatcgggaa atgattgagg
     4381 aaagactaaa aacatacgct cacctgttcg acgataaggt tatgaaacag ttaaagaggc
     4441 gtcgctatac gggctgggga cgattgtcgc ggaaacttat caacgggata agagacaagc
     4501 aaagtggtaa aactattctc gattttctaa agagcgacgg cttcgccaat aggaacttta
     4561 tgcagctgat ccatgatgac tctttaacct tcaaagagga tatacaaaag gcacaggttt
     4621 ccggacaagg ggactcattg cacgaacata ttgcgaatct tgctggttcg ccagccatca
     4681 aaaagggcat actccagaca gtcaaagtag tggatgagct agttaaggtc atgggacgtc
     4741 acaaaccgga aaacattgta atcgagatgg cacgcgaaaa tcaaacgact cagaaggggc
     4801 aaaaaaacag tcgagagcgg atgaagagaa tagaagaggg tattaaagaa ctgggcagcc
     4861 agatcttaaa ggagcatcct gtggaaaata cccaattgca gaacgagaaa ctttacctct
     4921 attacctaca aaatggaagg gacatgtatg ttgatcagga actggacata aaccgtttat
     4981 ctgattacga cgtcgatgcc attgtacccc aatccttttt gaaggacgat tcaatcgaca
     5041 ataaagtgct tacacgctcg gataagaacc gagggaaaag tgacaatgtt ccaagcgagg
     5101 aagtcgtaaa gaaaatgaag aactattggc ggcagctcct aaatgcgaaa ctgataacgc
     5161 aaagaaagtt cgataactta actaaagctg agaggggtgg cttgtctgaa cttgacaagg
     5221 ccggatttat taaacgtcag ctcgtggaaa cccgccaaat cacaaagcat gttgcacaga
     5281 tactagattc ccgaatgaat acgaaatacg acgagaacga taagctgatt cgggaagtca
     5341 aagtaatcac tttaaagtca aaattggtgt cggacttcag aaaggatttt caattctata
     5401 aagttaggga gataaataac taccaccatg cgcacgacgc ttatcttaat gccgtcgtag
     5461 ggaccgcact cattaagaaa tacccgaagc tagaaagtga gtttgtgtat ggtgattaca
     5521 aagtttatga cgtccgtaag atgatcgcga aaagcgaaca ggagataggc aaggctacag
     5581 ccaaatactt cttttattct aacattatga atttctttaa gacggaaatc actctggcaa
     5641 acggagagat acgcaaacga cctttaattg aaaccaatgg ggagacaggt gaaatcgtat
     5701 gggataaggg ccgggacttc gcgacggtga gaaaagtttt gtccatgccc caagtcaaca
     5761 tagtaaagaa aactgaggtg cagaccggag ggttttcaaa ggaatcgatt cttccaaaaa
     5821 ggaatagtga taagctcatc gctcgtaaaa aggactggga cccgaaaaag tacggtggct
     5881 tcgatagccc tacagttgcc tattctgtcc tagtagtggc aaaagttgag aagggaaaat
     5941 ccaagaaact gaagtcagtc aaagaattat tggggataac gattatggag cgctcgtctt
     6001 ttgaaaagaa ccccatcgac ttccttgagg cgaaaggtta caaggaagta aaaaaggatc
     6061 tcataattaa actaccaaag tatagtctgt ttgagttaga aaatggccga aaacggatgt
     6121 tggctagcgc cggagagctt caaaagggga acgaactcgc actaccgtct aaatacgtga
     6181 atttcctgta tttagcgtcc cattacgaga agttgaaagg ttcacctgaa gataacgaac
     6241 agaagcaact ttttgttgag cagcacaaac attatctcga cgaaatcata gagcaaattt
     6301 cggaattcag taagagagtc atcctagctg atgccaatct ggacaaagta ttaagcgcat
     6361 acaacaagca cagggataaa cccatacgtg agcaggcgga aaatattatc catttgttta
     6421 ctcttaccaa cctcggcgct ccagccgcat tcaagtattt tgacacaacg atagatcgca
     6481 aacgatacac ttctaccaag gaggtgctag acgcgacact gattcaccaa tccatcacgg
     6541 gattatatga aactcggata gatttgtcac agcttggggg tgacggatcc cccaagaaga
     6601 agaggaaagt ctcgagcgac tacaaagacc atgacggtga ttataaagat catgacatcg
     6661 attacaagga tgacgatgac aaggctgcag gaggcggtgg ataaccggtc atcatcacca
     6721 tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     6781 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     6841 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     6901 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     6961 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct cgataccgtc
     7021 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     7081 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctagggtgc
     7141 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     7201 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     7261 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     7321 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     7381 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7441 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7501 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7561 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7621 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     7681 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7741 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7801 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7861 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     7921 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7981 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     8041 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     8101 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     8161 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca g
//
