LOCUS       sequence-380249-       13012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral vector expressing sgRNA-1 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-1 lentiCRISPR v2 plasmid
FEATURES             Location/Qualifiers
     source          1..13012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(59..79)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        64..393
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      244..379
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     306..325
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        441..488
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             507..881
                     /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    1011..1132
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1048..1067)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1102..1121
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1181..1197)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1181..1197)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1194..1216)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1205..1221
                     /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(1229..1235,1236..1253,1254..1259))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1274..1295
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1412..1429)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1583..2171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1663..1682)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2342..3133,3134..3202))
                     /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     2965..2984
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3203..3307)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3382..3401)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3573..3952
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3954..4152
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4107..4127
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4170..4350
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4397..4522
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5015..5248
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5433..5477
                     /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    5775..5892
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5943..6191
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5943..5963
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6114..6133
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6212..6287
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        6349..6560
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6579..6588
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6585..10688
                     /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             6585..10688
                     /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             10689..10736
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             10737..10760
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             10770..10826
                     /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             10827..11423
                     /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     11320..11340
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    11439..12027
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11492..11512)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11910..11921)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             12099..12332
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(12358..12375)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12364..12588
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(12634..13012,1..50)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12721..12740)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12931..12952
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
       61 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
      121 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
      181 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
      241 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
      301 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
      361 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
      421 ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat
      481 aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca
      541 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg
      601 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
      661 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg
      721 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc
      781 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg
      841 ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg
      901 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
      961 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     1021 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     1081 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     1141 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     1201 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     1261 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     1321 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     1381 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     1441 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1501 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1561 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1621 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1681 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1741 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1801 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1861 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1921 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1981 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2041 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2101 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2161 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2221 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2281 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2341 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2401 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     2461 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     2521 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     2581 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     2641 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     2701 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     2761 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     2821 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     2881 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     2941 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3001 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3061 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3121 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3181 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3241 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3301 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     3361 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     3421 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     3481 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     3541 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
     3601 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     3661 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     3721 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     3781 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     3841 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     3901 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     3961 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     4021 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     4081 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     4141 ctatataagc agcgcgtttt gcctgtactg ggtctctctg gttagaccag atctgagcct
     4201 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     4261 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     4321 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
     4381 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     4441 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     4501 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     4561 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     4621 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     4681 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     4741 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     4801 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     4861 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     4921 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     4981 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     5041 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     5101 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     5161 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     5221 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     5281 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     5341 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     5401 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     5461 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     5521 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     5581 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     5641 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     5701 gatccattcg attagtgaac ggatcggcac tgcgtgcgcc aattctgcag acaaatggca
     5761 gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     5821 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     5881 attcaaaatt ttcgggttta ttacagggac agcagagatc cagtttggtt aattaaggta
     5941 ccgagggcct atttcccatg attccttcat atttgcatat acgatacaag gctgttagag
     6001 agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta
     6061 gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc
     6121 atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag
     6181 gacgaaacac cgggcggagc agctgaaaat cgttttagag ctagaaatag caagttaaaa
     6241 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttgaattcg
     6301 ctagctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca
     6361 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgatcc ggtgcctaga
     6421 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     6481 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     6541 ggtttgccgc cagaacacag gaccggttct agagcgctgc caccatggac aagaagtaca
     6601 gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca
     6661 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga
     6721 acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga
     6781 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct
     6841 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc
     6901 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg
     6961 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca
     7021 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg
     7081 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca
     7141 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg
     7201 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga
     7261 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc
     7321 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc
     7381 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt
     7441 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc
     7501 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg
     7561 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga
     7621 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg
     7681 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca
     7741 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg
     7801 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc
     7861 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct
     7921 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga
     7981 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg
     8041 cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga
     8101 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca
     8161 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa
     8221 aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag
     8281 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc
     8341 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact
     8401 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt
     8461 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca
     8521 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc
     8581 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg
     8641 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag
     8701 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca
     8761 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg
     8821 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag
     8881 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag
     8941 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc
     9001 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc
     9061 aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct
     9121 ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca
     9181 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc
     9241 tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag
     9301 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc
     9361 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga
     9421 atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
     9481 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg
     9541 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa
     9601 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg
     9661 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt
     9721 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa
     9781 acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag
     9841 tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca
     9901 gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact
     9961 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg
    10021 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga
    10081 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg
    10141 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc
    10201 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac
    10261 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga
    10321 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc
    10381 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta
    10441 atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg
    10501 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt
    10561 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca
    10621 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg
    10681 gaggcgacaa gcgacctgcc gccacaaaga aggctggaca ggctaagaag aagaaagatt
    10741 acaaagacga tgacgataag ggatccggcg caacaaactt ctctctgctg aaacaagccg
    10801 gagatgtcga agagaatcct ggaccgaccg agtacaagcc cacggtgcgc ctcgccaccc
    10861 gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
    10921 cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct
    10981 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
    11041 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
    11101 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
    11161 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcgga gtctcgcccg
    11221 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
    11281 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
    11341 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
    11401 tgacccgcaa gcccggtgcc tgaacgcgtt aagtcgacaa tcaacctctg gattacaaaa
    11461 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
    11521 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
    11581 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
    11641 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
    11701 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
    11761 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
    11821 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
    11881 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
    11941 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
    12001 ggatctccct ttgggccgcc tccccgcgtc gactttaaga ccaatgactt acaaggcagc
    12061 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
    12121 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
    12181 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
    12241 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
    12301 acccttttag tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct
    12361 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
    12421 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
    12481 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
    12541 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
    12601 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
    12661 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
    12721 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
    12781 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
    12841 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
    12901 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
    12961 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tc
//
