LOCUS       sequence-380259-       12823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral vector expressing sgRNA-3 targeting eIF3 binding site of 
            RPS4X 5'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12823)
  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 12823)
  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: RPS4X sgRNA-3 lentiCRISPR v2-Blast plasmid
FEATURES             Location/Qualifiers
     source          1..12823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(527..1318,1319..1387))
                     /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     1150..1169
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1388..1492)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1567..1586)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1758..2137
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2139..2337
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2292..2312
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2355..2535
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2582..2707
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3200..3433
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3618..3662
                     /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    3960..4077
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4128..4376
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4128..4148
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4299..4318
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4397..4472
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4534..4745
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4764..4773
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4770..8873
                     /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             4770..8873
                     /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             8874..8921
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8922..8945
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8955..9011
                     /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             9012..9410
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9435..10023
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9488..9508)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9906..9917)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10095..10328
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10354..10371)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10360..10584
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10630..11058
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10717..10736)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10927..10948
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11067..11087)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11072..11401
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11252..11387
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11314..11333
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11449..11496
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11515..11889
                     /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    12019..12140
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12056..12075)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12110..12129
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12189..12205)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12189..12205)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12202..12224)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12213..12229
                     /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(12237..12243,12244..12261,12262..12267))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12282..12303
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12420..12437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(12591..12823,1..356))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12671..12690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
       61 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
      121 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
      181 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
      241 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
      301 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
      361 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
      421 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
      481 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
      541 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
      601 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
      661 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
      721 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
      781 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
      841 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
      901 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
      961 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     1021 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     1081 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     1141 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     1201 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     1261 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     1321 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     1381 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     1441 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     1501 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     1561 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     1621 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     1681 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
     1741 ggccagatat acgcgttgac attgattatt gactagttat taatagtaat caattacggg
     1801 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     1861 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     1921 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     1981 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     2041 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     2101 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     2161 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     2221 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     2281 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagcgc
     2341 gttttgcctg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     2401 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     2461 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     2521 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     2581 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     2641 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     2701 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     2761 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     2821 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     2881 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     2941 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     3001 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca
     3061 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
     3121 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
     3181 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
     3241 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
     3301 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
     3361 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
     3421 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
     3481 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
     3541 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
     3601 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
     3661 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
     3721 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
     3781 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca
     3841 ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag
     3901 tgaacggatc ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt catccacaat
     3961 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
     4021 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg
     4081 gtttattaca gggacagcag agatccagtt tggttaatta aggtaccgag ggcctatttc
     4141 ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata attagaatta
     4201 atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc
     4261 ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa
     4321 cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccgtac
     4381 gcgcaccgga agacgggttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
     4441 atcaacttga aaaagtggca ccgagtcggt gcttttttga attcgctagc taggtcttga
     4501 aaggagtggg aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc
     4561 ccgagaagtt ggggggaggg gtcggcaatt gatccggtgc ctagagaagg tggcgcgggg
     4621 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac
     4681 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa
     4741 cacaggaccg gttctagagc gctgccacca tggacaagaa gtacagcatc ggcctggaca
     4801 tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga
     4861 aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc
     4921 tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa
     4981 gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg
     5041 ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata
     5101 agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga
     5161 agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc
     5221 tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg
     5281 agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga
     5341 cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca
     5401 tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg
     5461 gcgagaagaa gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca
     5521 acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct
     5581 acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc
     5641 tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg
     5701 agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg
     5761 acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt
     5821 tcttcgacca gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag
     5881 agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg
     5941 tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc
     6001 cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc
     6061 cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact
     6121 acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg
     6181 aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct
     6241 tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc
     6301 acagcctgct gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga
     6361 ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc
     6421 tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga
     6481 aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc
     6541 tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg
     6601 aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga
     6661 tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc
     6721 tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc
     6781 gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca
     6841 gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag
     6901 cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc
     6961 ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga
     7021 tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc
     7081 agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc
     7141 tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc
     7201 tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca
     7261 accggctgtc cgactacgat gtggaccata tcgtgcctca gagctttctg aaggacgact
     7321 ccatcgacaa caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc
     7381 cctccgaaga ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc
     7441 tgattaccca gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac
     7501 tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg
     7561 tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc
     7621 gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc
     7681 agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg
     7741 ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg
     7801 gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca
     7861 aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta
     7921 ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg
     7981 agatcgtgtg ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc
     8041 aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc
     8101 tgcccaagag gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt
     8161 acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa
     8221 agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa
     8281 gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga
     8341 aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga
     8401 agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca
     8461 aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg
     8521 ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg
     8581 agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc
     8641 tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc
     8701 acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca
     8761 tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga
     8821 gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggc gacaagcgac
     8881 ctgccgccac aaagaaggct ggacaggcta agaagaagaa agattacaaa gacgatgacg
     8941 ataagggatc cggcgcaaca aacttctctc tgctgaaaca agccggagat gtcgaagaga
     9001 atcctggacc gatggccaag cctttgtctc aagaagaatc caccctcatt gaaagagcaa
     9061 cggctacaat caacagcatc cccatctctg aagactacag cgtcgccagc gcagctctct
     9121 ctagcgacgg ccgcatcttc actggtgtca atgtatatca ttttactggg ggaccttgtg
     9181 cagaactcgt ggtgctgggc actgctgctg ctgcggcagc tggcaacctg acttgtatcg
     9241 tcgcgatcgg aaatgagaac aggggcatct tgagcccctg cggacggtgc cgacaggtgc
     9301 ttctcgatct gcatcctggg atcaaagcca tagtgaagga cagtgatgga cagccgacgg
     9361 cagttgggat tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa gaattcgata
     9421 tcaagcttat cggtaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     9481 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     9541 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     9601 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     9661 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     9721 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     9781 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     9841 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     9901 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     9961 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
    10021 cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
    10081 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc
    10141 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
    10201 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
    10261 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
    10321 ctctagcagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
    10381 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
    10441 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
    10501 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
    10561 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
    10621 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
    10681 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
    10741 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
    10801 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
    10861 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
    10921 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
    10981 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
    11041 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
    11101 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
    11161 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
    11221 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
    11281 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
    11341 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
    11401 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa
    11461 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc
    11521 aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc
    11581 tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc
    11641 cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc
    11701 ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg
    11761 tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg
    11821 gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag
    11881 caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
    11941 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
    12001 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
    12061 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
    12121 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
    12181 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
    12241 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
    12301 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
    12361 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
    12421 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
    12481 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
    12541 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
    12601 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
    12661 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
    12721 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
    12781 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcg
//
