LOCUS       sequence-235374-       14074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pLex_Cas9 is a single insert lentiviral vector expressing Cas9 
            driven by CMV promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14074)
  AUTHORS   Rubin AJ, Parker KR, Satpathy AT, Qi Y, Wu B, Ong AJ, Mumbach MR, Ji
            AL, Kim DS, Cho SW, Zarnegar BJ, Greenleaf WJ, Chang HY, Khavari PA
  TITLE     Coupled Single-Cell CRISPR Screening and Epigenomic Profiling 
            Reveals Causal Gene Regulatory Networks.
  JOURNAL   Cell. 2018 Dec 11. pii: S0092-8674(18)31514-9. doi: 
            10.1016/j.cell.2018.11.022.
  PUBMED    30580963
REFERENCE   2  (bases 1 to 14074)
  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: "Cell. 2018 
            Dec 11. pii: S0092-8674(18)31514-9. doi: 
            10.1016/j.cell.2018.11.022."
COMMENT     Sequence Label: pLex_Cas9
FEATURES             Location/Qualifiers
     source          1..14074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    652..777
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1274..1507
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1692..1736
                     /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    2034..2151
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(2216..2590)
                     /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"
     regulatory      2587..2596
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2600..2618)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2610..2657)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     enhancer        2762..3065
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3066..3269
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3220..3240
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3266..3290
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3358..7461
                     /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             3358..7461
                     /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             7462..7509
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7510..7533
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7543..7599
                     /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             7600..7998
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8020..8608
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8073..8093)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8491..8502)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8815..9048
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9149..9166)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9155..9379
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9425..9853
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9512..9531)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9722..9743
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9862..9882)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9867..10196
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10047..10182
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10109..10128
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             10245..11270
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             10245..11270
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    11400..11521
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11570..11586)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11570..11586)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11583..11605)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11594..11610
                     /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(11618..11624,11625..11642,11643..11648))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11663..11684
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11801..11818)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11972..12560)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12052..12071)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12731..13522,13523..13591))
                     /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     13354..13373
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13592..13696)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    13744..13878
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             join(14046..14074,1..605)
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac
       61 tacacaccag ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta
      121 ccagttgagc cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac
      181 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac
      241 agccgcctag catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc
      301 tgatatcgag cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg
      361 ggcgggactg gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg
      421 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
      481 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
      541 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
      601 tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga
      661 cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt
      721 acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt
      781 attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag
      841 aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt
      901 aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca
      961 tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc aaccctctat
     1021 tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa
     1081 gagcaaaaca aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg
     1141 gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa
     1201 ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa
     1261 gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg
     1321 gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc
     1381 agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct
     1441 ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac
     1501 agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga
     1561 atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt
     1621 gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa
     1681 accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga
     1741 attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag
     1801 gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg
     1861 gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg
     1921 aaggaataga agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg
     1981 gatcggcact gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa
     2041 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
     2101 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat
     2161 tacagggaca gcagagatcc agtttggtta gtaccgggcc cgctctagtc cggaatcagt
     2221 cctgctcctc ggccacgaag tgcacgcagt tgccggccgg gtcgcgcagg gcgaactccc
     2281 gcccccacgg ctgctcgccg atctcggtca tggccggccc ggaggcgtcc cggaagttcg
     2341 tggacacgac ctccgaccac tcggcgtaca gctcgtccag gccgcgcacc cacacccagg
     2401 ccagggtgtt gtccggcacc acctggtcct ggaccgcgct gatgaacagg gtcacgtcgt
     2461 cccggaccac accggcgaag tcgtcctcca cgaagtcccg ggagaacccg agccggtcgg
     2521 tccagaactc gaccgctccg gcgacgtcgc gcgcggtgag caccggaacg gcactggtca
     2581 acttggccat ggtggccctc ctatagtgag tcgtattata ctatgccgat atactatgcc
     2641 gatgattaat tgtcaacacg tgctgcaggt ccgaggttct agacgtatta ccgccatgca
     2701 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     2761 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     2821 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     2881 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     2941 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     3001 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     3061 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     3121 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     3181 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     3241 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac
     3301 gccatccacg ctgttttgac ctccatagaa gacaccgact ctactagagg atccaccatg
     3361 gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc
     3421 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
     3481 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
     3541 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
     3601 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
     3661 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
     3721 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
     3781 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
     3841 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
     3901 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
     3961 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
     4021 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg
     4081 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
     4141 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
     4201 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
     4261 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
     4321 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
     4381 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
     4441 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
     4501 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
     4561 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
     4621 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
     4681 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
     4741 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
     4801 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
     4861 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
     4921 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
     4981 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
     5041 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
     5101 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
     5161 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
     5221 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
     5281 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
     5341 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
     5401 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
     5461 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
     5521 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
     5581 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
     5641 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
     5701 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
     5761 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
     5821 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc
     5881 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac
     5941 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
     6001 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
     6061 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
     6121 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
     6181 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
     6241 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
     6301 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
     6361 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
     6421 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
     6481 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
     6541 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc
     6601 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
     6661 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc
     6721 agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat
     6781 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
     6841 gagctactgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
     6901 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
     6961 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag
     7021 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
     7081 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
     7141 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
     7201 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc
     7261 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
     7321 gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag
     7381 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
     7441 ctgtctcagc tgggaggcga caagcgacct gccgccacaa agaaggctgg acaggctaag
     7501 aagaagaaag attacaaaga cgatgacgat aagggatccg gcgcaacaaa cttctctctg
     7561 ctgaaacaag ccggagatgt cgaagagaat cctggaccga tggccaagcc tttgtctcaa
     7621 gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa
     7681 gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat
     7741 gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct
     7801 gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg
     7861 agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata
     7921 gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt
     7981 tatgtgtggg agggctaact cgagtctaga ggggttaaca atcaacctct ggattacaaa
     8041 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     8101 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     8161 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     8221 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     8281 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     8341 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     8401 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     8461 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     8521 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     8581 cggatctccc tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa
     8641 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag
     8701 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa
     8761 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag
     8821 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     8881 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     8941 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     9001 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     9061 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagaggc
     9121 cttgacattg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     9181 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     9241 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     9301 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     9361 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     9421 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     9481 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     9541 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     9601 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     9661 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     9721 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     9781 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     9841 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     9901 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     9961 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
    10021 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
    10081 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
    10141 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
    10201 tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgatgaaa aagcctgaac
    10261 tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga
    10321 tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat
    10381 atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc
    10441 actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga
    10501 gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa
    10561 ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg
    10621 atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta
    10681 catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga
    10741 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg
    10801 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga
    10861 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc
    10921 aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga
    10981 cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata
    11041 tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg
    11101 cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc
    11161 gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg
    11221 ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag cacgtgctac
    11281 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
    11341 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
    11401 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
    11461 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
    11521 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
    11581 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
    11641 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
    11701 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
    11761 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
    11821 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
    11881 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
    11941 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
    12001 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
    12061 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
    12121 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
    12181 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
    12241 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
    12301 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
    12361 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
    12421 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
    12481 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
    12541 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
    12601 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
    12661 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
    12721 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
    12781 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
    12841 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
    12901 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
    12961 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
    13021 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
    13081 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
    13141 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
    13201 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
    13261 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
    13321 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
    13381 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
    13441 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
    13501 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
    13561 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
    13621 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
    13681 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
    13741 atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
    13801 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
    13861 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc
    13921 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat
    13981 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta
    14041 gtagttggaa gggctaattc actcccaaag aaga
//
