LOCUS       sequence-380251-       13012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral vector expressing sgRNA-2 targeting eIF3 binding site of 
            RPS4X 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: RPS4X sgRNA-2 lentiCRISPR v2 plasmid
FEATURES             Location/Qualifiers
     source          1..13012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(16..33)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(187..775)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(267..286)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(946..1737,1738..1806))
                     /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     1569..1588
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1807..1911)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1986..2005)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2177..2556
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2558..2756
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2711..2731
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2774..2954
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3001..3126
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3619..3852
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4037..4081
                     /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    4379..4496
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4547..4795
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4547..4567
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4718..4737
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4816..4891
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4953..5164
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5183..5192
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5189..9292
                     /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             5189..9292
                     /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             9293..9340
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9341..9364
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9374..9430
                     /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             9431..10027
                     /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     9924..9944
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10043..10631
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10096..10116)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10514..10525)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10703..10936
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10962..10979)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10968..11192
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11238..11666
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11325..11344)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11535..11556
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11675..11695)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11680..12009
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11860..11995
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11922..11941
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12057..12104
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12123..12497
                     /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    12627..12748
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12664..12683)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12718..12737
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12797..12813)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12797..12813)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12810..12832)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12821..12837
                     /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(12845..12851,12852..12869,12870..12875))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12890..12911
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
       61 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
      121 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      181 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      241 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      301 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      361 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      421 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      481 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      541 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      601 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
      661 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      721 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
      781 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
      841 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
      901 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
      961 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1021 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1081 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1141 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1201 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1261 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1321 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1381 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1441 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1501 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1561 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1621 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1681 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1741 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     1801 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     1861 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     1921 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     1981 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     2041 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     2101 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
     2161 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
     2221 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     2281 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     2341 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     2401 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     2461 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     2521 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     2581 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     2641 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     2701 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg
     2761 ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     2821 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     2881 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     2941 gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag
     3001 ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga
     3061 ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag
     3121 agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca
     3181 gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga
     3241 ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag
     3301 ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca
     3361 accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag
     3421 atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag
     3481 acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt
     3541 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga
     3601 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac
     3661 tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt
     3721 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac
     3781 agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga
     3841 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc
     3901 ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat
     3961 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc
     4021 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt
     4081 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt
     4141 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag
     4201 gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag
     4261 gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt
     4321 gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt
     4381 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
     4441 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg
     4501 tttattacag ggacagcaga gatccagttt ggttaattaa ggtaccgagg gcctatttcc
     4561 catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa
     4621 tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct
     4681 tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac
     4741 ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccgagtc
     4801 ccggcgcgcc agcaagtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
     4861 tcaacttgaa aaagtggcac cgagtcggtg cttttttgaa ttcgctagct aggtcttgaa
     4921 aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
     4981 cgagaagttg gggggagggg tcggcaattg atccggtgcc tagagaaggt ggcgcggggt
     5041 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
     5101 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
     5161 acaggaccgg ttctagagcg ctgccaccat ggacaagaag tacagcatcg gcctggacat
     5221 cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa
     5281 attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct
     5341 gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag
     5401 atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc
     5461 caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa
     5521 gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa
     5581 gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct
     5641 gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga
     5701 gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac
     5761 ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat
     5821 cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg
     5881 cgagaagaag aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa
     5941 cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta
     6001 cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct
     6061 ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga
     6121 gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga
     6181 cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt
     6241 cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga
     6301 gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt
     6361 gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc
     6421 ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc
     6481 attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta
     6541 cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga
     6601 aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt
     6661 catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca
     6721 cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac
     6781 cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct
     6841 gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa
     6901 aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct
     6961 gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga
     7021 aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat
     7081 gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct
     7141 gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg
     7201 ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag
     7261 aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc
     7321 ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc
     7381 cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat
     7441 gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca
     7501 gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct
     7561 gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct
     7621 gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa
     7681 ccggctgtcc gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc
     7741 catcgacaac aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc
     7801 ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct
     7861 gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact
     7921 ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt
     7981 ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg
     8041 ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca
     8101 gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc
     8161 cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg
     8221 cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa
     8281 ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac
     8341 cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga
     8401 gatcgtgtgg gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca
     8461 agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct
     8521 gcccaagagg aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta
     8581 cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa
     8641 gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag
     8701 aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa
     8761 aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa
     8821 gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa
     8881 atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga
     8941 taatgagcag aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga
     9001 gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct
     9061 gtccgcctac aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca
     9121 cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat
     9181 cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag
     9241 catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acaagcgacc
     9301 tgccgccaca aagaaggctg gacaggctaa gaagaagaaa gattacaaag acgatgacga
     9361 taagggatcc ggcgcaacaa acttctctct gctgaaacaa gccggagatg tcgaagagaa
     9421 tcctggaccg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     9481 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     9541 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     9601 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     9661 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     9721 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     9781 caaggagccc gcgtggttcc tggccaccgt cggagtctcg cccgaccacc agggcaaggg
     9841 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     9901 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     9961 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
    10021 tgcctgaacg cgttaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
    10081 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
    10141 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
    10201 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
    10261 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
    10321 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
    10381 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
    10441 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
    10501 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
    10561 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
    10621 cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac
    10681 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
    10741 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
    10801 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
    10861 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
    10921 tggaaaatct ctagcagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
    10981 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
    11041 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
    11101 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
    11161 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
    11221 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
    11281 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
    11341 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
    11401 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
    11461 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
    11521 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
    11581 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
    11641 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
    11701 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
    11761 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
    11821 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
    11881 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
    11941 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
    12001 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg
    12061 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
    12121 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
    12181 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
    12241 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
    12301 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
    12361 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
    12421 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
    12481 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
    12541 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
    12601 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
    12661 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
    12721 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
    12781 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
    12841 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
    12901 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
    12961 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gg
//
