LOCUS       sequence-244571-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeting human MAPKAP1 (SIN1) gene by CRISPR-Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  AUTHORS   Hwang Y, Kim LC, Song W, Edwards DN, Cook RS, Chen J
  TITLE     Disruption of the Scaffolding Function of mLST8 Selectively Inhibits
            mTORC2 Assembly and Function and Suppresses mTORC2-Dependent Tumor 
            Growth In Vivo.
  JOURNAL   Cancer Res. 2019 Jul 1;79(13):3178-3184. doi: 
            10.1158/0008-5472.CAN-18-3658. Epub 2019 May 13.
  PUBMED    31085701
REFERENCE   2  (bases 1 to 13013)
  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: 
            "10.1158/0008-5472.CAN-18-3658"; journalName: "Cancer Res"; date: 
            "2019-07-1- 1"; volume: "79"; issue: "13"; pages: "3178-3184"
COMMENT     Sequence Label: sgMAPKAP1_2
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    89..206
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        257..505
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     257..277
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     428..447
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        527..602
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        664..875
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      894..903
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             900..5003
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             900..5003
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5004..5051
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5052..5075
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5085..5141
                     /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             5142..5738
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     5635..5655
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5754..6342
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5807..5827)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6225..6236)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6414..6647
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6673..6690)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6679..6903
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6949..7377
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7036..7055)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7246..7267
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7386..7406)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7391..7720
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7571..7706
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7633..7652
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7768..7815
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7834..8208
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    8338..8459
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8375..8394)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8429..8448
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8508..8524)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8508..8524)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8521..8543)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8532..8548
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(8556..8562,8563..8580,8581..8586))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8601..8622
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8739..8756)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8910..9498)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8990..9009)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9669..10460,10461..10529))
                     /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     10292..10311
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10530..10634)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10709..10728)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        10900..11279
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11281..11479
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11434..11454
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             11497..11677
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11724..11849
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12342..12575
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12760..12804
                     /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"
ORIGIN
        1 gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct
       61 gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca
      121 gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa
      181 aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt
      241 ggttaattaa ggtaccgagg gcctatttcc catgattcct tcatatttgc atatacgata
      301 caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca
      361 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
      421 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
      481 tatcttgtgg aaaggacgaa acaccgtgac acgggaatgt gtgagagttt tagagctaga
      541 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
      601 gcttttttga attcgctagc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
      661 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
      721 gatccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
      781 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
      841 ttctttttcg caacgggttt gccgccagaa cacaggaccg gttctagagc gctgccacca
      901 tggacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga
      961 tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc
     1021 acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg
     1081 ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct
     1141 atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac
     1201 tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca
     1261 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga
     1321 aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca
     1381 tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg
     1441 tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca
     1501 tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac
     1561 ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc
     1621 tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg
     1681 atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc
     1741 agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc
     1801 tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta
     1861 tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc
     1921 agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg
     1981 gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg
     2041 aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga
     2101 agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg
     2161 ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg
     2221 agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca
     2281 gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag
     2341 tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga
     2401 acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt
     2461 ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga
     2521 gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg
     2581 tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct
     2641 ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta
     2701 tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc
     2761 tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc
     2821 acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca
     2881 ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg
     2941 atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca
     3001 gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc
     3061 acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag
     3121 tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga
     3181 tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa
     3241 tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg
     3301 tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg
     3361 atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata
     3421 tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg
     3481 acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga
     3541 actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga
     3601 ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc
     3661 tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca
     3721 ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca
     3781 agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact
     3841 accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt
     3901 accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga
     3961 tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca
     4021 acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc
     4081 ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg
     4141 ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc
     4201 agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg
     4261 ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct
     4321 attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga
     4381 aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact
     4441 ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt
     4501 actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc
     4561 agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc
     4621 actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac
     4681 agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga
     4741 tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc
     4801 ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc
     4861 ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag
     4921 aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg
     4981 acctgtctca gctgggaggc gacaagcgac ctgccgccac aaagaaggct ggacaggcta
     5041 agaagaagaa agattacaaa gacgatgacg ataagggatc cggcgcaaca aacttctctc
     5101 tgctgaaaca agccggagat gtcgaagaga atcctggacc gaccgagtac aagcccacgg
     5161 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     5221 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     5281 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     5341 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     5401 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     5461 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     5521 tcggagtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     5581 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     5641 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     5701 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaac gcgttaagtc gacaatcaac
     5761 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     5821 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     5881 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     5941 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     6001 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     6061 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     6121 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     6181 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     6241 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     6301 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcgtcgactt taagaccaat
     6361 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     6421 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt
     6481 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     6541 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     6601 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa
     6661 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     6721 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     6781 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     6841 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     6901 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     6961 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     7021 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     7081 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     7141 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     7201 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     7261 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     7321 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     7381 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     7441 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     7501 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     7561 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     7621 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     7681 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     7741 atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc
     7801 atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg
     7861 gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc
     7921 tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc
     7981 atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc
     8041 ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc
     8101 tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc
     8161 gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga
     8221 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     8281 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     8341 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     8401 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     8461 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     8521 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     8581 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     8641 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     8701 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     8761 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     8821 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     8881 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     8941 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     9001 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     9061 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     9121 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     9181 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     9241 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     9301 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     9361 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     9421 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     9481 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     9541 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     9601 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     9661 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     9721 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     9781 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     9841 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     9901 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     9961 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
    10021 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
    10081 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
    10141 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
    10201 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
    10261 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
    10321 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
    10381 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
    10441 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
    10501 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
    10561 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
    10621 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
    10681 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
    10741 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
    10801 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
    10861 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
    10921 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
    10981 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
    11041 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
    11101 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
    11161 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
    11221 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
    11281 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
    11341 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
    11401 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
    11461 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
    11521 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
    11581 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
    11641 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
    11701 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
    11761 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
    11821 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
    11881 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
    11941 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
    12001 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
    12061 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
    12121 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
    12181 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
    12241 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
    12301 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
    12361 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
    12421 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
    12481 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
    12541 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
    12601 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
    12661 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
    12721 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
    12781 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
    12841 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
    12901 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
    12961 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg aga
//
