LOCUS       sequence-380241-       13012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral vector expressing sgRNA-1 targeting eIF3 binding site of 
            RPS15A 5'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13012)
  AUTHORS   Duan H, Zhang S, Zarai Y, Ollinger R, Wu Y, Sun L, Hu C, He Y, Tian 
            G, Rad R, Kong X, Cheng Y, Tuller T, Wolf DA
  TITLE     eIF3 mRNA selectivity profiling reveals eIF3k as a cancer-relevant 
            regulator of ribosome content.
  JOURNAL   EMBO J. 2023 Jun 15;42(12):e112362. doi: 10.15252/embj.2022112362. 
            Epub 2023 May 8.
  PUBMED    37155573
REFERENCE   2  (bases 1 to 13012)
  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"; journalName: "EMBO J."; 
            date: "2023-06-15"; volume: "42(12):e112362. doi"; pages: " 
            10.15252/embj.2022112362. Epub 2023 May 8"
COMMENT     Sequence Label: RPS15A sgRNA-1 lentiCRISPR v2 plasmid
FEATURES             Location/Qualifiers
     source          1..13012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(168..756)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(248..267)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(927..1718,1719..1787))
                     /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     1550..1569
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1788..1892)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1967..1986)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2158..2537
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2539..2737
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2692..2712
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2755..2935
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2982..3107
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3600..3833
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4018..4062
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    4360..4477
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4528..4776
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4528..4548
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4699..4718
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4797..4872
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4934..5145
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5164..5173
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5170..9273
                     /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             5170..9273
                     /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             9274..9321
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9322..9345
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9355..9411
                     /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             9412..10008
                     /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     9905..9925
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10024..10612
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10077..10097)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10495..10506)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10684..10917
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10943..10960)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10949..11173
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11219..11647
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11306..11325)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11516..11537
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11656..11676)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11661..11990
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11841..11976
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11903..11922
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12038..12085
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12104..12478
                     /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    12608..12729
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12645..12664)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12699..12718
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12778..12794)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12778..12794)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12791..12813)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12802..12818
                     /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(12826..12832,12833..12850,12851..12856))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12871..12892
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(join(13009..13012,1..14))
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
       61 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
      121 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
      181 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      241 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      301 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      361 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      421 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      481 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
      541 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      601 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      661 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
      721 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
      781 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
      841 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
      901 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
      961 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     1021 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     1081 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1141 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1201 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1261 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     1321 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     1381 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     1441 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     1501 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     1561 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     1621 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     1681 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     1741 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     1801 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     1861 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     1921 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     1981 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     2041 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     2101 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
     2161 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     2221 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     2281 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2341 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     2401 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2461 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2521 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     2581 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     2641 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     2701 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
     2761 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     2821 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     2881 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     2941 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     3001 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     3061 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     3121 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     3181 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     3241 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     3301 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     3361 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     3421 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     3481 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     3541 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     3601 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     3661 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     3721 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     3781 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     3841 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     3901 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     3961 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     4021 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     4081 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     4141 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     4201 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     4261 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     4321 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     4381 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     4441 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     4501 agatccagtt tggttaatta aggtaccgag ggcctatttc ccatgattcc ttcatatttg
     4561 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag
     4621 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
     4681 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
     4741 ttggctttat atatcttgtg gaaaggacga aacaccgaga gtgctactca ccggcggttt
     4801 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
     4861 ccgagtcggt gcttttttga attcgctagc taggtcttga aaggagtggg aattggctcc
     4921 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     4981 gtcggcaatt gatccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     5041 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     5101 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggaccg gttctagagc
     5161 gctgccacca tggacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc
     5221 tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac
     5281 accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa
     5341 acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac
     5401 cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc
     5461 ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc
     5521 atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac
     5581 ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc
     5641 ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac
     5701 aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag
     5761 gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc
     5821 aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg
     5881 ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac
     5941 ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac
     6001 ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc
     6061 gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg
     6121 agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct
     6181 ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac
     6241 ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag
     6301 cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac
     6361 ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga
     6421 gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg
     6481 gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg
     6541 ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac
     6601 ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac
     6661 ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac
     6721 ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc
     6781 gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg
     6841 aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc
     6901 gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg
     6961 ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa
     7021 gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa
     7081 acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc
     7141 ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag
     7201 acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc
     7261 cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc
     7321 gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc
     7381 ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag
     7441 aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc
     7501 cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa
     7561 gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag
     7621 aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat
     7681 gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg
     7741 accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag
     7801 aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc
     7861 gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc
     7921 aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc
     7981 cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc
     8041 ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag
     8101 atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg
     8161 atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac
     8221 gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc
     8281 ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc
     8341 cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc
     8401 cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag
     8461 accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat
     8521 aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc
     8581 accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg
     8641 aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat
     8701 cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag
     8761 ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc
     8821 ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac
     8881 ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg
     8941 tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc
     9001 aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac
     9061 cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat
     9121 ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc
     9181 agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag
     9241 acacggatcg acctgtctca gctgggaggc gacaagcgac ctgccgccac aaagaaggct
     9301 ggacaggcta agaagaagaa agattacaaa gacgatgacg ataagggatc cggcgcaaca
     9361 aacttctctc tgctgaaaca agccggagat gtcgaagaga atcctggacc gaccgagtac
     9421 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     9481 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     9541 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     9601 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     9661 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     9721 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     9781 ctggccaccg tcggagtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     9841 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     9901 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     9961 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaac gcgttaagtc
    10021 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
    10081 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
    10141 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
    10201 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
    10261 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
    10321 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
    10381 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
    10441 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
    10501 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
    10561 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcgtcgactt
    10621 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
    10681 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
    10741 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
    10801 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
    10861 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaggg
    10921 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
    10981 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
    11041 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
    11101 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
    11161 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
    11221 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
    11281 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
    11341 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
    11401 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
    11461 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
    11521 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
    11581 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
    11641 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
    11701 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
    11761 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
    11821 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
    11881 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
    11941 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
    12001 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
    12061 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
    12121 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
    12181 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
    12241 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
    12301 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
    12361 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
    12421 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
    12481 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
    12541 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
    12601 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
    12661 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
    12721 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
    12781 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
    12841 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
    12901 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
    12961 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cg
//
