LOCUS       sequence-380247-       13012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral vector expressing sgRNA-2 targeting eIF3 binding site of 
            RPS15A 5'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13012)
  AUTHORS   Duan H, Zhang S, Zarai Y, Ollinger R, Wu Y, Sun L, Hu C, He Y, Tian 
            G, Rad R, Kong X, Cheng Y, Tuller T, Wolf DA
  TITLE     eIF3 mRNA selectivity profiling reveals eIF3k as a cancer-relevant 
            regulator of ribosome content.
  JOURNAL   EMBO J. 2023 Jun 15;42(12):e112362. doi: 10.15252/embj.2022112362. 
            Epub 2023 May 8.
  PUBMED    37155573
REFERENCE   2  (bases 1 to 13012)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "EMBO J."; 
            date: "2023-06-15"; volume: "42(12):e112362. doi"; pages: " 
            10.15252/embj.2022112362. Epub 2023 May 8"
COMMENT     Sequence Label: RPS15A sgRNA-2 lentiCRISPR v2 plasmid
FEATURES             Location/Qualifiers
     source          1..13012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(125..145)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        130..459
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      310..445
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     372..391
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        507..554
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             573..947
                     /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    1077..1198
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1114..1133)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1168..1187
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1247..1263)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1247..1263)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1260..1282)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1271..1287
                     /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(1295..1301,1302..1319,1320..1325))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1340..1361
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1478..1495)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1649..2237)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1729..1748)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2408..3199,3200..3268))
                     /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     3031..3050
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3269..3373)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3448..3467)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3639..4018
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4020..4218
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4173..4193
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4236..4416
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4463..4588
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5081..5314
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5499..5543
                     /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    5841..5958
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6009..6257
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6009..6029
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6180..6199
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6278..6353
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        6415..6626
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6645..6654
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6651..10754
                     /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             6651..10754
                     /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             10755..10802
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             10803..10826
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             10836..10892
                     /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             10893..11489
                     /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     11386..11406
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    11505..12093
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11558..11578)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11976..11987)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             12165..12398
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(12424..12441)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12430..12654
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(12700..13012,1..116)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12787..12806)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(12997..13012,1..6)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
       61 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
      121 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
      181 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
      241 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
      301 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
      361 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
      421 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
      481 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
      541 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
      601 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
      661 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
      721 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
      781 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
      841 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
      901 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
      961 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     1021 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     1081 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     1141 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     1201 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     1261 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     1321 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     1381 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     1441 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     1501 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     1561 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     1621 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1681 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1741 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1801 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1861 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1921 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1981 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2041 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2101 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2161 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2221 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2281 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2341 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2401 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2461 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     2521 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     2581 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     2641 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     2701 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     2761 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     2821 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     2881 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     2941 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3001 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3061 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3121 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3181 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3241 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3301 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3361 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     3421 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3481 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     3541 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     3601 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
     3661 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     3721 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     3781 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     3841 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     3901 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     3961 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     4021 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     4081 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     4141 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     4201 ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct
     4261 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     4321 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     4381 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     4441 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     4501 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     4561 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     4621 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     4681 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     4741 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     4801 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     4861 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     4921 gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     4981 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     5041 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     5101 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     5161 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     5221 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     5281 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     5341 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     5401 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     5461 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     5521 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     5581 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
     5641 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
     5701 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
     5761 gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa
     5821 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     5881 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     5941 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttggttaatt
     6001 aaggtaccga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
     6061 ttagagagat aattagaatt aatttgactg taaacacaaa gatattagta caaaatacgt
     6121 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
     6181 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
     6241 ggaaaggacg aaacaccgtt ccgccatctt tccgcgcgtt ttagagctag aaatagcaag
     6301 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg
     6361 aattcgctag ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt cagtgggcag
     6421 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgatccggtg
     6481 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     6541 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     6601 gcaacgggtt tgccgccaga acacaggacc ggttctagag cgctgccacc atggacaaga
     6661 agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg atcaccgacg
     6721 agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca
     6781 agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc
     6841 tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag
     6901 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt
     6961 ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg
     7021 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg
     7081 acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt
     7141 tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc
     7201 tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca
     7261 gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa
     7321 atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc
     7381 tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac
     7441 tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg
     7501 accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg
     7561 acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga
     7621 gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc
     7681 ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg
     7741 acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg
     7801 acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga
     7861 ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc
     7921 ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc
     7981 tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct
     8041 ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca
     8101 agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca
     8161 acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc
     8221 tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc
     8281 agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc
     8341 tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg
     8401 aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca
     8461 aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga
     8521 cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg
     8581 acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc
     8641 ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga
     8701 agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct
     8761 ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca
     8821 ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg
     8881 tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg
     8941 ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga
     9001 tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca
     9061 cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg
     9121 tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc
     9181 agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc
     9241 ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc
     9301 ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg
     9361 agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa
     9421 cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg
     9481 acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt
     9541 ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg
     9601 cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc
     9661 tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca
     9721 agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga
     9781 actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg
     9841 agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc
     9901 ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg
     9961 gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga
    10021 aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc
    10081 tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc
    10141 tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag
    10201 ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt
    10261 tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa
    10321 acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga
    10381 agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc
    10441 actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg
    10501 acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag
    10561 agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct
    10621 tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg
    10681 acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc
    10741 agctgggagg cgacaagcga cctgccgcca caaagaaggc tggacaggct aagaagaaga
    10801 aagattacaa agacgatgac gataagggat ccggcgcaac aaacttctct ctgctgaaac
    10861 aagccggaga tgtcgaagag aatcctggac cgaccgagta caagcccacg gtgcgcctcg
    10921 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
    10981 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
    11041 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
    11101 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
    11161 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
    11221 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggagtct
    11281 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
    11341 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
    11401 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
    11461 ggtgcatgac ccgcaagccc ggtgcctgaa cgcgttaagt cgacaatcaa cctctggatt
    11521 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
    11581 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
    11641 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
    11701 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
    11761 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
    11821 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
    11881 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
    11941 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
    12001 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
    12061 cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa
    12121 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
    12181 ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
    12241 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
    12301 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
    12361 cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat
    12421 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
    12481 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
    12541 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
    12601 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
    12661 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
    12721 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
    12781 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
    12841 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
    12901 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
    12961 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc ca
//
