LOCUS       sequence-259488-        8711 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses spCas9 and sgRNA scaffold with com aptamer replacing the 
            Tetraloop.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8711)
  AUTHORS   Lu Z, Yao X, Lyu P, Yadav M, Yoo K, Atala A, Lu B
  TITLE     Lentiviral Capsid-Mediated Streptococcus pyogenes Cas9 
            Ribonucleoprotein Delivery for Efficient and Safe Multiplex Genome 
            Editing.
  JOURNAL   CRISPR J. 2021 Mar 16. doi: 10.1089/crispr.2020.0106.
  PUBMED    33733873
REFERENCE   2  (bases 1 to 8711)
  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"; journalName: "CRISPR J. 
            2021 Mar 16. doi: 10.1089/crispr.2020.0106."
COMMENT     Sequence Label: pspCas9-3'UTR-Tetra-com-vector
FEATURES             Location/Qualifiers
     source          1..8711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          134..361
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      373..382
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             382..447
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             454..474
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             499..4599
                     /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             499..4599
                     /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             4600..4647
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4957..4974)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4963..5170
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5182..5322
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5182..5311
                     /label=AAV2 ITR
     rep_origin      5397..5852
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5484..5503)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5694..5715
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5869..5888)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5988..6010
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6048..6066)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6134..6238
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6239..6307,6308..7099)
                     /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(6457..6476)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7270..7858
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7759..7778
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7920..8168
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7920..7940
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8091..8110
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        8308..8593
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
       61 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
      121 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc
      181 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
      241 cggcccttct cctccgggct gtaattagct gagcaagagg taagggttta agggatggtt
      301 ggttggtggg gtattaatgt ttaattacct ggagcacctg cctgaaatca ctttttttca
      361 ggttggaccg gtgccaccat ggactataag gaccacgacg gagactacaa ggatcatgat
      421 attgattaca aagacgatga cgataagatg gccccaaaga agaagcggaa ggtcggtatc
      481 cacggagtcc cagcagccga caagaagtac agcatcggcc tggacatcgg caccaactct
      541 gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg
      601 ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc
      661 ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg
      721 aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac
      781 agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg
      841 caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc
      901 taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat
      961 ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac
     1021 cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg
     1081 ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga
     1141 ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat
     1201 ggcctgttcg gaaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac
     1261 ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg
     1321 gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac
     1381 ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc
     1441 cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg
     1501 aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc
     1561 aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc
     1621 atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga
     1681 gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac
     1741 ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac
     1801 aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg
     1861 gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc
     1921 tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg
     1981 accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac
     2041 gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga
     2101 aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc
     2161 aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc
     2221 gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac
     2281 gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt
     2341 ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg
     2401 ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga
     2461 tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc
     2521 ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag
     2581 ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc
     2641 cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag
     2701 ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag
     2761 cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag
     2821 aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc
     2881 ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac
     2941 ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac
     3001 tacgatgtgg accatatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag
     3061 gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc
     3121 gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga
     3181 aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc
     3241 ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg
     3301 gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg
     3361 atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg
     3421 cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc
     3481 gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg
     3541 tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag
     3601 tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc
     3661 gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat
     3721 aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg
     3781 aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac
     3841 agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac
     3901 agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag
     3961 aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag
     4021 aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc
     4081 atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc
     4141 tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc
     4201 ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa
     4261 cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag
     4321 ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac
     4381 aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg
     4441 accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg
     4501 tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg
     4561 tacgagacac ggatcgacct gtctcagctg ggaggcgaca aaaggccggc ggccacgaaa
     4621 aaggccggcc aggcaaaaaa gaaaaaggga tcctaagctc gctttcttgc tgtccaattt
     4681 ctattaaagg ttcctttgtt ccctaagtcc aactactaaa ctgggggata ttatgaaggg
     4741 ccttgagcat ctggattctg cctaataaaa aacatttatt ttcattgcta gctcgctttc
     4801 ttgctgtcca atttctatta aaggttcctt tgttccctaa gtccaactac taaactgggg
     4861 gatattatga agggccttga gcatctggat tctgcctaat aaaaaacatt tattttcatt
     4921 gcgaattcta gaaaattcct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     4981 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     5041 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     5101 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagag aatagcaggc
     5161 atgctgggga ggtacggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     5221 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     5281 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     5341 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     5401 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5461 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5521 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5581 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     5641 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     5701 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     5761 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5821 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     5881 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     5941 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     6001 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     6061 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     6121 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     6181 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     6241 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6301 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6361 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6421 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6481 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6541 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6601 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6661 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6721 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6781 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6841 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6901 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtggaagccg
     6961 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     7021 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     7081 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     7141 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     7201 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7261 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7321 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7381 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     7441 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7501 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7561 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7621 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     7681 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7741 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7801 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7861 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtg
     7921 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
     7981 taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
     8041 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata
     8101 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac
     8161 gaaacaccgg tgtcttcctc gaggaagacc cgtttgagag ctaggccctg aatgcctgcg
     8221 agcatcccac ggcctagcaa gttcaaataa ggctagtccg ttatcaactt gaaaaagtgg
     8281 caccgagtcg gtgctttttt tggtacccgt tacataactt acggtaaatg gcccgcctgg
     8341 ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag ggactttcca
     8401 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     8461 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcattg
     8521 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     8581 cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc
     8641 ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc
     8701 gggggggggg g
//
