LOCUS       sequence-244698-       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_1
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    269..386
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        437..685
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     437..457
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     608..627
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        707..782
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        844..1055
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      1074..1083
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1080..5183
                     /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             1080..5183
                     /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             5184..5231
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5232..5255
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5265..5321
                     /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             5322..5918
                     /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     5815..5835
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5934..6522
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5987..6007)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6405..6416)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6594..6827
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6853..6870)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6859..7083
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7129..7557
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7216..7235)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7426..7447
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7566..7586)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7571..7900
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7751..7886
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7813..7832
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7948..7995
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8014..8388
                     /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    8518..8639
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8555..8574)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8609..8628
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8688..8704)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8688..8704)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8701..8723)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8712..8728
                     /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(8736..8742,8743..8760,8761..8766))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8781..8802
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8919..8936)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9090..9678)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9170..9189)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9849..10640,10641..10709))
                     /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     10472..10491
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10710..10814)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10889..10908)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        11080..11459
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11461..11659
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11614..11634
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             11677..11857
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11904..12029
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12522..12755
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12940..12984
                     /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 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
       61 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
      121 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
      181 gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct
      241 gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca
      301 gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa
      361 aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt
      421 ggttaattaa ggtaccgagg gcctatttcc catgattcct tcatatttgc atatacgata
      481 caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca
      541 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
      601 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
      661 tatcttgtgg aaaggacgaa acaccgtgag gtaatatcga ctgactgttt tagagctaga
      721 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
      781 gcttttttga attcgctagc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
      841 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
      901 gatccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
      961 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
     1021 ttctttttcg caacgggttt gccgccagaa cacaggaccg gttctagagc gctgccacca
     1081 tggacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga
     1141 tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc
     1201 acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg
     1261 ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct
     1321 atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac
     1381 tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca
     1441 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga
     1501 aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca
     1561 tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg
     1621 tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca
     1681 tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac
     1741 ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc
     1801 tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg
     1861 atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc
     1921 agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc
     1981 tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta
     2041 tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc
     2101 agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg
     2161 gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg
     2221 aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga
     2281 agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg
     2341 ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg
     2401 agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca
     2461 gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag
     2521 tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga
     2581 acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt
     2641 ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga
     2701 gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg
     2761 tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct
     2821 ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta
     2881 tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc
     2941 tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc
     3001 acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca
     3061 ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg
     3121 atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca
     3181 gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc
     3241 acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag
     3301 tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga
     3361 tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa
     3421 tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg
     3481 tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg
     3541 atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata
     3601 tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg
     3661 acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga
     3721 actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga
     3781 ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc
     3841 tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca
     3901 ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca
     3961 agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact
     4021 accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt
     4081 accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga
     4141 tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca
     4201 acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc
     4261 ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg
     4321 ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc
     4381 agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg
     4441 ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct
     4501 attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga
     4561 aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact
     4621 ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt
     4681 actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc
     4741 agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc
     4801 actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac
     4861 agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga
     4921 tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc
     4981 ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc
     5041 ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag
     5101 aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg
     5161 acctgtctca gctgggaggc gacaagcgac ctgccgccac aaagaaggct ggacaggcta
     5221 agaagaagaa agattacaaa gacgatgacg ataagggatc cggcgcaaca aacttctctc
     5281 tgctgaaaca agccggagat gtcgaagaga atcctggacc gaccgagtac aagcccacgg
     5341 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     5401 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     5461 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     5521 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     5581 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     5641 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     5701 tcggagtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     5761 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     5821 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     5881 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaac gcgttaagtc gacaatcaac
     5941 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     6001 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     6061 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     6121 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     6181 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     6241 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     6301 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     6361 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     6421 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     6481 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcgtcgactt taagaccaat
     6541 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     6601 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt
     6661 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     6721 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     6781 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa
     6841 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     6901 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     6961 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     7021 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     7081 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     7141 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     7201 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     7261 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     7321 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     7381 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     7441 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     7501 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     7561 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     7621 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     7681 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     7741 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     7801 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     7861 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     7921 atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc
     7981 atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg
     8041 gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc
     8101 tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc
     8161 atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc
     8221 ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc
     8281 tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc
     8341 gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga
     8401 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     8461 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     8521 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     8581 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     8641 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     8701 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     8761 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     8821 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     8881 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     8941 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     9001 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     9061 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     9121 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     9181 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     9241 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     9301 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     9361 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     9421 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     9481 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     9541 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     9601 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     9661 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     9721 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     9781 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     9841 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     9901 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     9961 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
    10021 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
    10081 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
    10141 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
    10201 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
    10261 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
    10321 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
    10381 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
    10441 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
    10501 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
    10561 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
    10621 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
    10681 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
    10741 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
    10801 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
    10861 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
    10921 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
    10981 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
    11041 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
    11101 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
    11161 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
    11221 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
    11281 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
    11341 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
    11401 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
    11461 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
    11521 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
    11581 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
    11641 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
    11701 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
    11761 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
    11821 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
    11881 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
    11941 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
    12001 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
    12061 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
    12121 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
    12181 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
    12241 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
    12301 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
    12361 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
    12421 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
    12481 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
    12541 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
    12601 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
    12661 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
    12721 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
    12781 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
    12841 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
    12901 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
    12961 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gct
//
