LOCUS       sequence-245340-       11005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of spCas9 with blasticidin resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11005)
  AUTHORS   Tarumoto Y, Lin S, Wang J, Milazzo JP, Xu Y, Lu B, Yang Z, Wei Y, 
            Polyanskaya S, Wunderlich M, Gray NS, Stegmaier K, Vakoc CR
  TITLE     Salt-Inducible Kinase inhibition suppresses acute myeloid leukemia 
            progression in vivo.
  JOURNAL   Blood. 2019 Nov 1. pii: 422697. doi: 10.1182/blood.2019001576.
  PUBMED    31697837
REFERENCE   2  (bases 1 to 11005)
  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: "Blood. 2019
            Nov 1. pii: 422697. doi: 10.1182/blood.2019001576."
COMMENT     Sequence Label: LentiV_Cas9_Blast
FEATURES             Location/Qualifiers
     source          1..11005
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             232..412
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1077..1310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1495..1539
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1723..1840
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1909..2120
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2140..2149
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2149..2172
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2179..2199
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2224..6324
                     /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             2224..6324
                     /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             6325..6372
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6388..6444
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             6445..6843
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    6862..7450
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6915..6935)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7333..7344)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7522..7755
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7833..7954
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7870..7889)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7924..7943
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7994..8129
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8149..8168)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8150..8168)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8178..8195)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8178..8194)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8187..8209)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8336..8791
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8423..8442)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8633..8654
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8817..8921
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8922..8990,8991..9782)
                     /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(9140..9159)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9953..10541
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10442..10461
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10695..10712
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10829..10850
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10865..10870,10871..10888,10889..10895)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10903..10919
                     /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)."
     primer_bind     10908..10930
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10927..10943
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10927..10943
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10962..10982
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10964..10982
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagggga cccagaccac aattttaaaa gaaaaggggg
     1741 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     1801 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     1861 cagagatcca ctttggcgcc ggctcgagtg gctccggtgc ccgtcagtgg gcagagcgca
     1921 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
     1981 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     2041 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     2101 ggtttgccgc cagaacacag gtgtcgtgac gcgggatccg ccaccatgga ttacaaagac
     2161 gatgacgata agatggcccc aaagaagaag cggaaggtcg gtatccacgg agtcccagca
     2221 gccgacaaga agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg
     2281 atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg
     2341 cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag
     2401 gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc
     2461 tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga
     2521 ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc
     2581 aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag
     2641 aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac
     2701 atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac
     2761 gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc
     2821 atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga
     2881 cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac
     2941 ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag
     3001 gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc
     3061 cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc
     3121 ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct
     3181 atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg
     3241 cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc
     3301 ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg
     3361 gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg
     3421 aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac
     3481 gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc
     3541 gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc
     3601 agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa
     3661 gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag
     3721 aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg
     3781 tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg
     3841 agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc
     3901 gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc
     3961 tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt
     4021 atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg
     4081 ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc
     4141 cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc
     4201 aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg
     4261 gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac
     4321 agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg
     4381 cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca
     4441 gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg
     4501 atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga
     4561 atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc
     4621 gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg
     4681 gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat
     4741 atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc
     4801 gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag
     4861 aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg
     4921 accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag
     4981 ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac
     5041 actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc
     5101 aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac
     5161 taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag
     5221 taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag
     5281 atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc
     5341 aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg
     5401 cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt
     5461 gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg
     5521 cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc
     5581 gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc
     5641 tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg
     5701 aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac
     5761 tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag
     5821 tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg
     5881 cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc
     5941 cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa
     6001 cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg
     6061 atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag
     6121 cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc
     6181 cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa
     6241 gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc
     6301 gacctgtctc agctgggagg cgacaagcgt cctgctgcta ctaagaaagc tggtcaagct
     6361 aagaaaaaga aagctagcgg cagcggcgcc accaacttca gcctgctgaa gcaggccggc
     6421 gacgtggagg agaaccccgg ccccatggcc aagcctttgt ctcaagaaga atccaccctc
     6481 attgaaagag caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc
     6541 agcgcagctc tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact
     6601 gggggacctt gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac
     6661 ctgacttgta tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg
     6721 tgccgacagg tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat
     6781 ggacagccga cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc
     6841 taatgaacgc gttaagtcga caatcaacct ctggattaca aaatttgtga aagattgact
     6901 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     6961 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     7021 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     7081 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     7141 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     7201 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     7261 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     7321 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     7381 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     7441 gcctccccgc gtcgacttta agaccaatga cttacaaggc agctgtagat cttagccact
     7501 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc
     7561 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     7621 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     7681 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     7741 ggaaaatctc tagcagtacg tatagtagtt catgtcatct tattattcag tatttataac
     7801 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     7861 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     7921 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     7981 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     8041 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     8101 gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag
     8161 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     8221 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     8281 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
     8341 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     8401 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     8461 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     8521 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     8581 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     8641 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     8701 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     8761 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     8821 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     8881 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     8941 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     9001 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     9061 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     9121 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     9181 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     9241 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     9301 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     9361 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     9421 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     9481 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     9541 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     9601 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     9661 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     9721 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     9781 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     9841 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     9901 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     9961 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
    10021 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
    10081 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
    10141 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
    10201 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
    10261 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
    10321 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
    10381 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
    10441 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
    10501 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
    10561 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
    10621 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
    10681 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
    10741 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
    10801 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
    10861 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
    10921 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
    10981 gggaacaaaa gctggagctg caagc
//
