LOCUS       sequence-261312-       12503 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 12503)
  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 12503)
  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-T2-pGK-Pur
FEATURES             Location/Qualifiers
     source          1..12503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    98..122
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        135..383
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     135..155
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     306..325
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        404..479
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        576..861
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        863..1140
                     /label=chicken beta-actin promoter
     intron          1141..1368
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1380..1389
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1389..1454
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1461..1481
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1506..5606
                     /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             1506..5606
                     /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             5607..5654
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     promoter        5703..6202
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5738..5761)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6135..6154
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6223..6822
                     /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(6223..6242)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6719..6739
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(6827..6851)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6898..6987
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     misc_feature    7054..7642
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7107..7127)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7525..7536)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7645..7661)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7646..7662)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    7898..7931
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             7945..8125
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(8187..8775)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8267..8286)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8946..9737,9738..9806))
                     /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     9569..9588
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9807..9911)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9986..10005)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        10177..10556
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        10558..10756
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     10711..10731
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             10774..10954
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11001..11126
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11623..11856
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12041..12085
                     /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    12383..12500
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gtttattaca gggacagcag agatccagtt tggttagtac cgggcccgct ctagccctag
       61 atttagcttg ccacaaccca caaggagacg accttccaca agtttgtaca aaaaagttgg
      121 cggatccacc atgtgagggc ctatttccca tgattccttc atatttgcat atacgataca
      181 aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa
      241 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
      301 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
      361 tcttgtggaa aggacgaaac accgaaacag acatggccgg cgagttttag agctagaaat
      421 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
      481 tttttgtttt agagctagaa atagcaagtt aaaataaggc tagtccgttt ttagcgcgtg
      541 cgccaattct gcagacaaat ggctctagag gtacccgtta cataacttac ggtaaatggc
      601 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caatagtaac gccaataggg
      661 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      721 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      781 tggcattgtg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      841 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
      901 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
      961 atgggggcgg gggggggggg ggggcgcgcg ccaggsgggg cggggcgggg cgaggggcgg
     1021 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
     1081 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
     1141 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
     1201 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
     1261 ccgggctgta attagctgag caagaggtaa gggtttaagg gatggttggt tggtggggta
     1321 ttaatgttta attacctgga gcacctgcct gaaatcactt tttttcaggt tggaccggtg
     1381 ccaccatgga ctataaggac cacgacggag actacaagga tcatgatatt gattacaaag
     1441 acgatgacga taagatggcc ccaaagaaga agcggaaggt cggtatccac ggagtcccag
     1501 cagccgacaa gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg
     1561 tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc
     1621 ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg
     1681 aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct
     1741 gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca
     1801 gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg
     1861 gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa
     1921 agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc
     1981 acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg
     2041 acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc
     2101 ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca
     2161 gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa
     2221 acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg
     2281 aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg
     2341 cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca
     2401 tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct
     2461 ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc
     2521 ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg
     2581 ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc
     2641 tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc
     2701 ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc
     2761 acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga
     2821 tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca
     2881 gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg
     2941 aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata
     3001 agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg
     3061 tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc
     3121 tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga
     3181 ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa
     3241 tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa
     3301 ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg
     3361 tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg
     3421 cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg
     3481 gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc
     3541 tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg
     3601 acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc
     3661 tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga
     3721 cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg
     3781 tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga
     3841 gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc
     3901 ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc
     3961 gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc
     4021 atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa
     4081 gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga
     4141 agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc
     4201 tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac
     4261 agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga
     4321 acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt
     4381 ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca
     4441 actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa
     4501 agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga
     4561 agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca
     4621 gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc
     4681 ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt
     4741 ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg
     4801 tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga
     4861 tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg
     4921 cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg
     4981 tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg
     5041 actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta
     5101 agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac
     5161 tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca
     5221 gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg
     5281 aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag
     5341 tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata
     5401 agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag
     5461 cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca
     5521 aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga
     5581 tcgacctgtc tcagctggga ggcgacaaaa ggccggcggc cacgaaaaag gccggccagg
     5641 caaaaaagaa aaagtaagaa ttcgtccagc caagcttagg atctcgacct cgaaattcta
     5701 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct
     5761 gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag
     5821 gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt
     5881 caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc
     5941 acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct
     6001 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag
     6061 gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc
     6121 ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc
     6181 ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc
     6241 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     6301 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     6361 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     6421 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     6481 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     6541 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     6601 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     6661 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     6721 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     6781 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaggtacccc aactttcttg
     6841 tacaaagtgg tgggtggagg cggttcaggt ggcggaggaa gcggaggcgg tggaagttac
     6901 ccctacgacg tgcccgacta cgccggctat ccgtatgatg tcccggacta tgcaggatcc
     6961 tatccatacg acgttccaga ttacgcttag ttaattaaac atgtttaagg gttccggttc
     7021 cactaggtac aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt
     7081 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     7141 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     7201 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     7261 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     7321 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     7381 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     7441 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     7501 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     7561 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     7621 tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gatcgagacc tagaaaaaca
     7681 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc
     7741 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac
     7801 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     7861 aattcactcc caacgaagac aagatctgct ttttgctata acttcgtata atgtatgcta
     7921 tacgaagtta ttccggactg tactgggtct ctctggttag accagatctg agcctgggag
     7981 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     8041 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     8101 tagtcagtgt ggaaaatctc tagcagcatg tgagcaaaag gccagcaaaa ggccaggaac
     8161 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     8221 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     8281 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     8341 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     8401 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     8461 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     8521 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     8581 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     8641 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     8701 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     8761 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     8821 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     8881 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     8941 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     9001 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     9061 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     9121 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     9181 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     9241 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     9301 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     9361 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     9421 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     9481 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     9541 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     9601 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     9661 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     9721 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     9781 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     9841 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     9901 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     9961 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
    10021 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
    10081 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
    10141 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt
    10201 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
    10261 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
    10321 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
    10381 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
    10441 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
    10501 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
    10561 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
    10621 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
    10681 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
    10741 aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga ccagatctga
    10801 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
    10861 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
    10921 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
    10981 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
    11041 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
    11101 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
    11161 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
    11221 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
    11281 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
    11341 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
    11401 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
    11461 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
    11521 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
    11581 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
    11641 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
    11701 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
    11761 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
    11821 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
    11881 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
    11941 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
    12001 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
    12061 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
    12121 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
    12181 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
    12241 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
    12301 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
    12361 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
    12421 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
    12481 ttacaaaaat tcaaaatttt cgg
//
