LOCUS       sequence-244572-       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_3
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    320..437
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        488..736
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     488..508
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     659..678
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        758..833
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        895..1106
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      1125..1134
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1131..5234
                     /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             1131..5234
                     /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             5235..5282
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5283..5306
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5316..5372
                     /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             5373..5969
                     /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     5866..5886
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5985..6573
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6038..6058)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6456..6467)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6645..6878
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6904..6921)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6910..7134
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7180..7608
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7267..7286)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7477..7498
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7617..7637)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7622..7951
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7802..7937
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7864..7883
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7999..8046
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8065..8439
                     /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    8569..8690
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8606..8625)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8660..8679
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8739..8755)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8739..8755)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8752..8774)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8763..8779
                     /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(8787..8793,8794..8811,8812..8817))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8832..8853
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8970..8987)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9141..9729)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9221..9240)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9900..10691,10692..10760))
                     /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     10523..10542
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10761..10865)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10940..10959)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        11131..11510
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11512..11710
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11665..11685
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             11728..11908
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11955..12080
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12573..12806
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             join(12991..13013,1..22)
                     /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 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
       61 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
      121 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
      181 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
      241 agacagatcc attcgattag tgaacggatc ggcactgcgt gcgccaattc tgcagacaaa
      301 tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg
      361 aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta
      421 caaaaattca aaattttcgg gtttattaca gggacagcag agatccagtt tggttaatta
      481 aggtaccgag ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt
      541 tagagagata attagaatta atttgactgt aaacacaaag atattagtac aaaatacgtg
      601 acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga
      661 ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg
      721 gaaaggacga aacaccgccc gatcaggtcc tgcacccgtt ttagagctag aaatagcaag
      781 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg
      841 aattcgctag ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt cagtgggcag
      901 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgatccggtg
      961 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     1021 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     1081 gcaacgggtt tgccgccaga acacaggacc ggttctagag cgctgccacc atggacaaga
     1141 agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg atcaccgacg
     1201 agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca
     1261 agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc
     1321 tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag
     1381 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt
     1441 ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg
     1501 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg
     1561 acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt
     1621 tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc
     1681 tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca
     1741 gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa
     1801 atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc
     1861 tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac
     1921 tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg
     1981 accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg
     2041 acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga
     2101 gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc
     2161 ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg
     2221 acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg
     2281 acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga
     2341 ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc
     2401 ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc
     2461 tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct
     2521 ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca
     2581 agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca
     2641 acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc
     2701 tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc
     2761 agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc
     2821 tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg
     2881 aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca
     2941 aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga
     3001 cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg
     3061 acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc
     3121 ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga
     3181 agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct
     3241 ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca
     3301 ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg
     3361 tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg
     3421 ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga
     3481 tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca
     3541 cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg
     3601 tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc
     3661 agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc
     3721 ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc
     3781 ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg
     3841 agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa
     3901 cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg
     3961 acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt
     4021 ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg
     4081 cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc
     4141 tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca
     4201 agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga
     4261 actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg
     4321 agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc
     4381 ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg
     4441 gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga
     4501 aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc
     4561 tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc
     4621 tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag
     4681 ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt
     4741 tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa
     4801 acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga
     4861 agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc
     4921 actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg
     4981 acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag
     5041 agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct
     5101 tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg
     5161 acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc
     5221 agctgggagg cgacaagcga cctgccgcca caaagaaggc tggacaggct aagaagaaga
     5281 aagattacaa agacgatgac gataagggat ccggcgcaac aaacttctct ctgctgaaac
     5341 aagccggaga tgtcgaagag aatcctggac cgaccgagta caagcccacg gtgcgcctcg
     5401 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     5461 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     5521 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     5581 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     5641 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     5701 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggagtct
     5761 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     5821 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     5881 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     5941 ggtgcatgac ccgcaagccc ggtgcctgaa cgcgttaagt cgacaatcaa cctctggatt
     6001 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     6061 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     6121 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     6181 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     6241 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     6301 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     6361 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     6421 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     6481 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     6541 cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa
     6601 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     6661 ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
     6721 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     6781 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     6841 cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat
     6901 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     6961 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     7021 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     7081 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     7141 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     7201 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     7261 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     7321 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     7381 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     7441 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     7501 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     7561 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     7621 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     7681 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     7741 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     7801 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     7861 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     7921 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     7981 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     8041 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc
     8101 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
     8161 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
     8221 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
     8281 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
     8341 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
     8401 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
     8461 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     8521 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     8581 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     8641 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     8701 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     8761 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     8821 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     8881 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     8941 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     9001 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     9061 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     9121 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     9181 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     9241 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     9301 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     9361 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     9421 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     9481 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     9541 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     9601 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     9661 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     9721 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     9781 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     9841 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     9901 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     9961 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
    10021 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
    10081 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
    10141 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
    10201 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
    10261 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
    10321 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
    10381 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
    10441 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
    10501 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
    10561 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
    10621 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
    10681 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
    10741 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
    10801 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
    10861 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
    10921 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
    10981 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
    11041 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
    11101 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
    11161 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
    11221 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
    11281 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
    11341 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
    11401 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
    11461 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
    11521 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
    11581 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
    11641 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
    11701 atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
    11761 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
    11821 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
    11881 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
    11941 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
    12001 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
    12061 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
    12121 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
    12181 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
    12241 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
    12301 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
    12361 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
    12421 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
    12481 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
    12541 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
    12601 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
    12661 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
    12721 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
    12781 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
    12841 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
    12901 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
    12961 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt gga
//
