LOCUS       sequence-380255-       12823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral vector expressing sgRNA-4 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-4 lentiCRISPR v2-Blast plasmid
FEATURES             Location/Qualifiers
     source          1..12823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(91..108)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(262..850)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(342..361)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1021..1812,1813..1881))
                     /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     1644..1663
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1882..1986)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2061..2080)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2252..2631
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2633..2831
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2786..2806
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2849..3029
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3076..3201
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3694..3927
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4112..4156
                     /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    4454..4571
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4622..4870
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4622..4642
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4793..4812
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4891..4966
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        5028..5239
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5258..5267
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5264..9367
                     /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             5264..9367
                     /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             9368..9415
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9416..9439
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9449..9505
                     /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             9506..9904
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9929..10517
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9982..10002)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10400..10411)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10589..10822
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10848..10865)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10854..11078
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11124..11552
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11211..11230)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11421..11442
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11561..11581)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11566..11895
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11746..11881
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11808..11827
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11943..11990
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12009..12383
                     /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    12513..12634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12550..12569)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12604..12623
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12683..12699)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12683..12699)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12696..12718)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12707..12723
                     /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(12731..12737,12738..12755,12756..12761))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12776..12797
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
       61 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
      121 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
      181 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
      241 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      301 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
      361 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
      421 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
      481 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
      541 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
      601 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
      661 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
      721 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
      781 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
      841 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
      901 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
      961 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1021 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1081 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     1141 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     1201 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     1261 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     1321 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     1381 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     1441 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     1501 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     1561 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     1621 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     1681 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     1741 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     1801 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     1861 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     1921 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     1981 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     2041 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     2101 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     2161 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     2221 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag
     2281 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     2341 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     2401 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     2461 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     2521 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     2581 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     2641 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     2701 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     2761 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     2821 tatataagca gcgcgttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg
     2881 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     2941 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     3001 cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa
     3061 gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga
     3121 ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag
     3181 agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa
     3241 attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc
     3301 agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga
     3361 caaatactgg gacagctaca accatccctt cagacaggat cagaagaact tagatcatta
     3421 tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag
     3481 gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg
     3541 gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata
     3601 taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag
     3661 agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg
     3721 agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt
     3781 attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca
     3841 tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga
     3901 aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg
     3961 caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa
     4021 tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct taatacactc
     4081 cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga
     4141 taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt
     4201 attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat
     4261 agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc
     4321 gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag
     4381 atccattcga ttagtgaacg gatcggcact gcgtgcgcca attctgcaga caaatggcag
     4441 tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
     4501 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa
     4561 ttcaaaattt tcgggtttat tacagggaca gcagagatcc agtttggtta attaaggtac
     4621 cgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
     4681 gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
     4741 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
     4801 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
     4861 acgaaacacc gctgcgttag gcaaggaaag gttttagagc tagaaatagc aagttaaaat
     4921 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgaattcgc
     4981 tagctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac
     5041 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg gtgcctagag
     5101 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     5161 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     5221 gtttgccgcc agaacacagg accggttcta gagcgctgcc accatggaca agaagtacag
     5281 catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa
     5341 ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa
     5401 cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag
     5461 aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt
     5521 cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag agtccttcct
     5581 ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt
     5641 ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac
     5701 cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg
     5761 ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat
     5821 ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt
     5881 ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat
     5941 cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct
     6001 gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct
     6061 gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta
     6121 cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct
     6181 gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga
     6241 cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa
     6301 gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg
     6361 agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac
     6421 cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga
     6481 caacggcagc atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca
     6541 ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt
     6601 ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac
     6661 cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc
     6721 ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa
     6781 ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa
     6841 agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa
     6901 ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga
     6961 ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg
     7021 gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt
     7081 cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt
     7141 tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa
     7201 agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct
     7261 gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga
     7321 cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga
     7381 ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa
     7441 tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga
     7501 gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga
     7561 gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga
     7621 gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct
     7681 gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca
     7741 ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt
     7801 tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa
     7861 gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct
     7921 gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg
     7981 cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca
     8041 gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa
     8101 tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt
     8161 ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga
     8221 cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag
     8281 cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga
     8341 gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt
     8401 caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa
     8461 cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt
     8521 gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag
     8581 caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg
     8641 ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt
     8701 ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat
     8761 caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg
     8821 ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct
     8881 ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact
     8941 ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa
     9001 gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct
     9061 ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa
     9121 tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc
     9181 cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta
     9241 ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac
     9301 cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg
     9361 aggcgacaag cgacctgccg ccacaaagaa ggctggacag gctaagaaga agaaagatta
     9421 caaagacgat gacgataagg gatccggcgc aacaaacttc tctctgctga aacaagccgg
     9481 agatgtcgaa gagaatcctg gaccgatggc caagcctttg tctcaagaag aatccaccct
     9541 cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc
     9601 cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac
     9661 tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa
     9721 cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg
     9781 gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga
     9841 tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg
     9901 ctaagaattc gatatcaagc ttatcggtaa tcaacctctg gattacaaaa tttgtgaaag
     9961 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
    10021 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
    10081 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
    10141 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
    10201 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
    10261 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
    10321 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
    10381 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
    10441 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
    10501 ttgggccgcc tccccgcgtc gactttaaga ccaatgactt acaaggcagc tgtagatctt
    10561 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
    10621 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
    10681 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
    10741 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
    10801 tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
    10861 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
    10921 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
    10981 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
    11041 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
    11101 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
    11161 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
    11221 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
    11281 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
    11341 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
    11401 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
    11461 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
    11521 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
    11581 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
    11641 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
    11701 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
    11761 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
    11821 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
    11881 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
    11941 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
    12001 actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg
    12061 gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact
    12121 tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg
    12181 tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt
    12241 ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg
    12301 gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact
    12361 tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct
    12421 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
    12481 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
    12541 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
    12601 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
    12661 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
    12721 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
    12781 tgagctaact cacattaatt gcgttgcgct cactgcccgc ttt
//
