LOCUS       sequence-210930-       12506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression CRISPR/Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12506)
  AUTHORS   Lu G, Middleton RE, Sun H, Naniong M, Ott CJ, Mitsiades CS, Wong KK,
            Bradner JE, Kaelin WG Jr
  TITLE     The myeloma drug lenalidomide promotes the cereblon-dependent 
            destruction of Ikaros proteins.
  JOURNAL   Science. 2014 Jan 17;343(6168):305-9. doi: 10.1126/science.1244917. 
            Epub 2013 Nov 29.
  PUBMED    24292623
REFERENCE   2  (bases 1 to 12506)
  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: 
            "10.1126/science.1244917"; journalName: "Science"; date: 
            "2014-01-17- 17"; volume: "343"; issue: "6168"; pages: "305-9"
COMMENT     Sequence Label: plenti-px330-CRBN-T1-pGK-Pur
FEATURES             Location/Qualifiers
     source          1..12506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    111..135
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        148..396
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     148..168
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     319..338
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        417..492
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        587..872
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        874..1151
                     /label=chicken beta-actin promoter
     intron          1152..1380
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1392..1401
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1405..1470
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1477..1497
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1522..5622
                     /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             1522..5622
                     /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             5623..5670
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     promoter        5719..6218
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5754..5777)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6151..6170
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6239..6838
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6239..6258)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6735..6755
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(6843..6867)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6914..7003
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     misc_feature    7070..7658
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7123..7143)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7541..7552)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7661..7677)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7662..7678)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    7914..7947
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             7961..8141
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(8203..8791)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8283..8302)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8962..9753,9754..9822))
                     /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     9585..9604
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9823..9927)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10002..10021)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        10193..10572
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        10574..10772
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     10727..10747
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             10790..10970
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11017..11142
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11639..11872
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12057..12101
                     /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    join(12399..12506,1..10)
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag taccgggccc
       61 gctctagccc tagatttagc ttgccacaac ccacaaggag acgaccttcc acaagtttgt
      121 acaaaaaagt tggcggatcc accatgtgag ggcctatttc ccatgattcc ttcatatttg
      181 catatacgat acaaggctgt tagagagata attggaatta atttgactgt aaacacaaag
      241 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
      301 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
      361 ttggctttat atatcttgtg gaaaggacga aacaccgtcc tgctgatctc cttcgcgttt
      421 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
      481 ccgagtcggt gcttttttgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg
      541 tttttagcgc gtgcgccaat tctgcagaca aggctctaga ggtacccgtt acataactta
      601 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaatagtaa
      661 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      721 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      781 aatggcccgc ctggcattgt gcccagtaca tgaccttatg ggactttcct acttggcagt
      841 acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca
      901 ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat
      961 tttgtgcagc gatgggggcg gggggggggg gggggcgcgc gccaggcggg gcggggcggg
     1021 gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc
     1081 tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg
     1141 cgcggcgggc gggagtcgct gcgacgctgc cttcgccccg tgccccgctc cgccgccgcc
     1201 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
     1261 ggcccttctc ctccgggctg taattagctg agcaagaggt aagggtttaa gggatggttg
     1321 gttggtgggg tattaatgtt taattacctg gagcacctgc ctgaaatcac tttttttcag
     1381 gttggaccgg tgccaccatg gatggactat aaggaccacg acggagacta caaggatcat
     1441 gatattgatt acaaagacga tgacgataag atggccccaa agaagaagcg gaaggtcggt
     1501 atccacggag tcccagcagc cgacaagaag tacagcatcg gcctggacat cggcaccaac
     1561 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg
     1621 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac
     1681 agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga
     1741 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac
     1801 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag
     1861 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc
     1921 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc
     1981 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg
     2041 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag
     2101 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc
     2161 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag
     2221 aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc
     2281 aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac
     2341 ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag
     2401 aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag
     2461 gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg
     2521 ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag
     2581 agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag
     2641 ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac
     2701 agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc
     2761 cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag
     2821 gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct
     2881 ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc
     2941 ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg
     3001 atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg
     3061 tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg
     3121 agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag
     3181 accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc
     3241 ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac
     3301 cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac
     3361 attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa
     3421 cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg
     3481 agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag
     3541 tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg
     3601 cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc
     3661 ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag
     3721 aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac
     3781 aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag
     3841 aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag
     3901 atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac
     3961 tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc
     4021 gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac
     4081 aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag
     4141 gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag
     4201 agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc
     4261 ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc
     4321 ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa
     4381 gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa
     4441 gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga
     4501 accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag
     4561 gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc
     4621 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac
     4681 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg
     4741 gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc
     4801 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg
     4861 aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc
     4921 gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc
     4981 aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc
     5041 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg
     5101 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg
     5161 gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac
     5221 ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag
     5281 aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc
     5341 gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac
     5401 aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc
     5461 ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag
     5521 aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc
     5581 ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg
     5641 aaaaaggccg gccaggcaaa aaagaaaaag taagaattcg tccagccaag cttaggatct
     5701 cgacctcgaa attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg
     5761 ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt
     5821 ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt
     5881 ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt
     5941 gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat
     6001 ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg
     6061 ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg
     6121 cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg
     6181 ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttaccat
     6241 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     6301 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     6361 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     6421 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     6481 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     6541 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     6601 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     6661 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     6721 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     6781 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagg
     6841 taccccaact ttcttgtaca aagtggtggg tggaggcggt tcaggtggcg gaggaagcgg
     6901 aggcggtgga agttacccct acgacgtgcc cgactacgcc ggctatccgt atgatgtccc
     6961 ggactatgca ggatcctatc catacgacgt tccagattac gcttagttaa ttaaacatgt
     7021 ttaagggttc cggttccact aggtacaatt cgatatcaag cttatcgata atcaacctct
     7081 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     7141 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     7201 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     7261 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     7321 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     7381 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     7441 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     7501 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     7561 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     7621 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgatc
     7681 gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt
     7741 gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac
     7801 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     7861 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gctataactt
     7921 cgtataatgt atgctatacg aagttattcc ggactgtact gggtctctct ggttagacca
     7981 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     8041 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     8101 atccctcaga cccttttagt cagtgtggaa aatctctagc agcatgtgag caaaaggcca
     8161 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     8221 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     8281 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     8341 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     8401 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     8461 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     8521 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     8581 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     8641 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     8701 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     8761 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     8821 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     8881 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     8941 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     9001 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     9061 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     9121 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     9181 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     9241 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     9301 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     9361 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     9421 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     9481 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     9541 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     9601 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     9661 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     9721 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     9781 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     9841 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     9901 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     9961 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
    10021 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
    10081 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
    10141 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
    10201 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
    10261 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
    10321 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
    10381 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
    10441 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
    10501 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
    10561 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
    10621 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
    10681 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
    10741 aggcgtgtac ggtgggaggt ctatataagc agcgcgtttt gcctgtactg ggtctctctg
    10801 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
    10861 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
    10921 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
    10981 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
    11041 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
    11101 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
    11161 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
    11221 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
    11281 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
    11341 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
    11401 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
    11461 aagaccaccg cacagcaagc ggccggccgc tgatcttcag acctggagga ggagatatga
    11521 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag
    11581 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag
    11641 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga
    11701 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc
    11761 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc
    11821 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt
    11881 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta
    11941 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta
    12001 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga
    12061 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa
    12121 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa
    12181 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat
    12241 cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag
    12301 aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg
    12361 cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt
    12421 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
    12481 gaattacaaa aacaaattac aaaaat
//
