LOCUS       sequence-380253-       12823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral vector expressing sgRNA-4 targeting eIF3 binding site of 
            RPS15A 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: RPS15A sgRNA-4 lentiCRISPR v2-Blast plasmid
FEATURES             Location/Qualifiers
     source          1..12823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(70..87)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(241..829)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(321..340)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1000..1791,1792..1860))
                     /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     1623..1642
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1861..1965)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2040..2059)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2231..2610
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2612..2810
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2765..2785
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2828..3008
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3055..3180
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3673..3906
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4091..4135
                     /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    4433..4550
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4601..4849
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4601..4621
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4772..4791
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4870..4945
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        5007..5218
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5237..5246
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5243..9346
                     /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             5243..9346
                     /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             9347..9394
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9395..9418
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9428..9484
                     /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             9485..9883
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9908..10496
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9961..9981)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10379..10390)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10568..10801
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10827..10844)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10833..11057
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11103..11531
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11190..11209)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11400..11421
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11540..11560)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11545..11874
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11725..11860
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11787..11806
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11922..11969
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11988..12362
                     /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    12492..12613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12529..12548)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12583..12602
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12662..12678)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12662..12678)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12675..12697)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12686..12702
                     /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(12710..12716,12717..12734,12735..12740))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12755..12776
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
       61 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
      121 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
      181 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
      241 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
      301 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      361 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      421 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      481 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      541 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      601 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      661 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      721 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      781 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      841 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      901 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      961 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     1021 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     1081 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     1141 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     1201 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1261 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     1321 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1381 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1441 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1501 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1561 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1621 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1681 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1741 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1801 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1861 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     1921 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     1981 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     2041 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     2101 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     2161 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     2221 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     2281 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     2341 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     2401 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     2461 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     2521 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     2581 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2641 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2701 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2761 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc
     2821 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     2881 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     2941 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     3001 ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct
     3061 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg
     3121 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc
     3181 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga
     3241 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
     3301 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
     3361 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
     3421 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
     3481 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg
     3541 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
     3601 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
     3661 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
     3721 cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
     3781 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
     3841 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
     3901 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
     3961 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg
     4021 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
     4081 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
     4141 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
     4201 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
     4261 atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga
     4321 aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg
     4381 atcggcactg cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa
     4441 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     4501 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     4561 acagggacag cagagatcca gtttggttaa ttaaggtacc gagggcctat ttcccatgat
     4621 tccttcatat ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac
     4681 tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta
     4741 gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa
     4801 gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg gattgcagga
     4861 tggcgccgag ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
     4921 tgaaaaagtg gcaccgagtc ggtgcttttt tgaattcgct agctaggtct tgaaaggagt
     4981 gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa
     5041 gttgggggga ggggtcggca attgatccgg tgcctagaga aggtggcgcg gggtaaactg
     5101 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
     5161 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagga
     5221 ccggttctag agcgctgcca ccatggacaa gaagtacagc atcggcctgg acatcggcac
     5281 caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa
     5341 ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt
     5401 cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac
     5461 cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt
     5521 ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca
     5581 cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc
     5641 caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct
     5701 gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga
     5761 cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa
     5821 ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc
     5881 tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa
     5941 gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa
     6001 gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga
     6061 cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc
     6121 caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac
     6181 caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac
     6241 cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga
     6301 ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta
     6361 caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct
     6421 gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca
     6481 gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct
     6541 gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg
     6601 ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat
     6661 caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga
     6721 gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct
     6781 gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg
     6841 aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt
     6901 caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga
     6961 gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac
     7021 ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga
     7081 ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga
     7141 ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg
     7201 gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa
     7261 gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt
     7321 catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt
     7381 gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat
     7441 taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg
     7501 gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg
     7561 acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag
     7621 ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct
     7681 gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct
     7741 gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga
     7801 caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga
     7861 agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac
     7921 ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa
     7981 ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca
     8041 gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt
     8101 gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta
     8161 caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt
     8221 gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta
     8281 caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac
     8341 cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc
     8401 caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt
     8461 gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa
     8521 tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa
     8581 gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg
     8641 cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa
     8701 gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag
     8761 cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga
     8821 cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat
     8881 gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt
     8941 gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga
     9001 gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat
     9061 cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc
     9121 ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt
     9181 taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg
     9241 gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac
     9301 cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacaagc gacctgccgc
     9361 cacaaagaag gctggacagg ctaagaagaa gaaagattac aaagacgatg acgataaggg
     9421 atccggcgca acaaacttct ctctgctgaa acaagccgga gatgtcgaag agaatcctgg
     9481 accgatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag caacggctac
     9541 aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc tctctagcga
     9601 cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt gtgcagaact
     9661 cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat
     9721 cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg tgcttctcga
     9781 tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga cggcagttgg
     9841 gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taagaattcg atatcaagct
     9901 tatcggtaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     9961 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
    10021 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
    10081 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
    10141 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
    10201 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
    10261 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
    10321 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
    10381 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
    10441 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg
    10501 actttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
    10561 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
    10621 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
    10681 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
    10741 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
    10801 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
    10861 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
    10921 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
    10981 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
    11041 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
    11101 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
    11161 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
    11221 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
    11281 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
    11341 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
    11401 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
    11461 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
    11521 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
    11581 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
    11641 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
    11701 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
    11761 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
    11821 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
    11881 ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg
    11941 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga
    12001 ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg
    12061 accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg
    12121 acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct
    12181 gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga
    12241 acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg
    12301 agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact
    12361 gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
    12421 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
    12481 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
    12541 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
    12601 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
    12661 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
    12721 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
    12781 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtg
//
