LOCUS       sequence-274134-       15524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of gRNA and dCas9-VP64 from the same plasmid for CRISPRa.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15524)
  TITLE     pLentiV2-dCas9-VP64
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 15524)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pLentiV2-dCas9-VP64
FEATURES             Location/Qualifiers
     source          1..15524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(84..104)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        89..418
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      269..404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     331..350
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        466..513
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             532..906
                     /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    1036..1157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1073..1092)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1127..1146
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1206..1222)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1206..1222)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1219..1241)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1230..1246
                     /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(1254..1260,1261..1278,1279..1284))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1299..1320
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1437..1454)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1608..2196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1688..1707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2367..3158,3159..3227))
                     /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     2990..3009
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3228..3332)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3407..3426)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3602..3981
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3983..4181
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4136..4156
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4199..4379
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4426..4551
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5044..5277
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5462..5506
                     /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    5804..5921
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5972..6220
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5972..5992
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6143..6162
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8102..8177
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8239..8450
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             8467..8514
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(8515..8538,8539..8541,8542..11097,11098..11100,
                     11101..12615)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             12670..12690
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12715..12864
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             12886..12942
                     /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             12943..13539
                     /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     13436..13456
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    13576..14164
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(13629..13649)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(14047..14058)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(14167..14183)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14168..14184)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             14689..14869
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(14895..14912)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    14901..15125
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(15171..15524,1..75)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(15258..15277)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     15468..15489
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
       61 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
      121 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
      181 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      241 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
      301 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
      361 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
      421 gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca
      481 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag
      541 ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg
      601 accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg
      661 gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg
      721 gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc
      781 acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg
      841 cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag
      901 gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
      961 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     1021 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1081 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1141 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     1201 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1261 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1321 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     1381 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     1441 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     1501 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     1561 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1621 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1681 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1741 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1801 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1861 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1921 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1981 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2041 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2101 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2161 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2221 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2281 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2341 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2401 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     2461 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     2521 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     2581 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     2641 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     2701 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     2761 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2821 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2881 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2941 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3001 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3061 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3121 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3181 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3241 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3301 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     3361 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     3421 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     3481 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     3541 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgcgcgt
     3601 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     3661 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     3721 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     3781 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     3841 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     3901 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     3961 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     4021 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     4081 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     4141 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg
     4201 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     4261 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     4321 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     4381 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
     4441 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
     4501 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
     4561 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
     4621 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
     4681 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
     4741 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
     4801 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
     4861 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga
     4921 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     4981 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     5041 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     5101 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     5161 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     5221 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     5281 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     5341 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     5401 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     5461 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     5521 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     5581 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     5641 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     5701 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     5761 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     5821 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     5881 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     5941 gcagagatcc agtttggtta attaaggtac cgagggccta tttcccatga ttccttcata
     6001 tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac
     6061 aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt
     6121 tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga
     6181 tttcttggct ttatatatct tgtggaaagg acgaaacacc ggagacggtt gtaaatgagc
     6241 acacaaaata cacatgctaa aatattatat tctatgacct ttataaaatc aaccaaaatc
     6301 ttctttttaa taactttagt atcaataatt agaattttta tgttcctttt tgcaaacttt
     6361 taataaaaat gagcaaaata aaaaaacgct agttttagta actcgcgttg ttttcttcac
     6421 ctttaataat agctactcca ccacttgttc ctaagcggtc agctcctgct tcaatcattt
     6481 tttgagcatc ttcaaatgtt ctaactccac cagctgcttt aactaaagca ttgtctttaa
     6541 caactgactt cattagttta acatcttcaa atgttgcacc tgattttgaa aatcctgttg
     6601 atgttttaac aaattctaat ccagcttcaa cagctatttc acaagctttc atgatttctt
     6661 cttttgttaa taaacaattt tccataatac atttaacaac atgtgatcca gctgcttttt
     6721 ttacagcttt catgtcttct aaaactaatt cataattttt gtcttttaat gcaccaatat
     6781 ttaataccat atcaatttct gttgcaccat ctttaattgc ttcagaaact tcgaatgctt
     6841 ttgtagctgt tgtgcatgca cctagaggaa aacctacaac atttgttatt cctacatttg
     6901 tgccttttaa taattcttta caatagcttg ttcaatatga attaacacaa actgttgcaa
     6961 aatcaaattc aattgcttca tcacataatt gtttaatttc agctttcgta gcatcttgtt
     7021 ttaataatgt gtgatctata tatttgttta gtttcatttt ttctcctata tattcatttt
     7081 taattttaat tctttaataa tttcgtctac tttaacttta gcgttttgaa cagattcacc
     7141 aacacctata aaataaattt ttagtttagg ttcagttcca cttgggcgaa cagcaaatca
     7201 tgacttatct tctaaataaa attttagtaa gtcttgtcct ggcatattat acattccatc
     7261 gatgtagtct tcaacattaa caactttaag tccagcaatt tgagttaagg gtgttgctct
     7321 caatgatttc attaatggtt caatttttaa tttcttttct tctggtttaa aattcaagtt
     7381 taaagtgaaa gtgtaatatg cacccatttc tttaaataaa tcttctaaat agtctactaa
     7441 tgttttattt tgttttttat aaaatcaagc agcctctgct attaatatag aagcttgtat
     7501 tccatcttta tctctagctg agtcatcaat tacatatcca taactttctt cataagcaaa
     7561 aacaaaattt aatccgttat cttcttcttt agcaatttct ctacccattc atttaaatcc
     7621 agttaaagtt tttacaatat taactccata tttttcatga gcgattctat cacccaaatc
     7681 acttgttaca aaacttgaat atagagccgg attttttgga atgctattta agcgttttag
     7741 atttgataat tttcaatcaa ttaaaattgg tcctgtttga tttccatcta atcttacaaa
     7801 atgaccatca tgttttattg ccattccaaa tctgtcagca tctgggtcat tcataataat
     7861 aatatctgca tcatgtttaa taccatattc aagcggtatt tttcatgcag gatcaaattc
     7921 tggatttgga tttacaacat ttttaaatgt ttcatcttca aatgcatgct cttcaacctc
     7981 aataacgtta tatcctgatt cacgtaatat ttttggggta aatttagttc ctgttccatt
     8041 aactgcgcta aaaataattt ttaaatcttt tttagcttct tgctcttttt tgtacgtctc
     8101 tgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
     8161 tggcaccgag tcggtgcttt tttgaattcg ctagctaggt cttgaaagga gtgggaattg
     8221 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     8281 gaggggtcgg caattgatcc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     8341 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     8401 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gcgtacggcc
     8461 accatgaaaa ggccggcggc cacgaaaaag gccggccagg caaaaaagaa aaaggacaag
     8521 aagtacagca tcggcctggc catcggcacc aactctgtgg gctgggccgt gatcaccgac
     8581 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
     8641 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg
     8701 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
     8761 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
     8821 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
     8881 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
     8941 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
     9001 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
     9061 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
     9121 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa
     9181 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggcaa cctgattgcc
     9241 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
     9301 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
     9361 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
     9421 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
     9481 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
     9541 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt
     9601 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
     9661 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg
     9721 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
     9781 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
     9841 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
     9901 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
     9961 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
    10021 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag
    10081 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
    10141 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
    10201 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
    10261 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
    10321 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
    10381 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
    10441 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
    10501 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
    10561 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
    10621 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
    10681 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
    10741 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
    10801 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
    10861 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
    10921 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
    10981 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca catcgtgcct
    11041 cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaaggcc
    11101 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
    11161 cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc
    11221 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
    11281 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
    11341 gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
    11401 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
    11461 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
    11521 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
    11581 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
    11641 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
    11701 gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg
    11761 cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
    11821 ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag
    11881 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
    11941 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
    12001 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
    12061 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
    12121 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga
    12181 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
    12241 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag
    12301 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
    12361 gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga
    12421 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
    12481 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
    12541 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct
    12601 cagctgggag gcgacagcgc tggaggaggt ggaagcggag gaggaggaag cggaggagga
    12661 ggtagcggac ctaagaaaaa gaggaaggtg gcggccgctg gatccggacg ggctgacgca
    12721 ttggacgatt ttgatctgga tatgctggga agtgacgccc tcgatgattt tgaccttgac
    12781 atgcttggtt cggatgccct tgatgacttt gacctcgaca tgctcggcag tgacgccctt
    12841 gatgatttcg acctggacat gctgattaac tgtacaggca gtggagcaac aaacttctct
    12901 ctgctgaaac aagccggaga tgtcgaagag aatcctggac cgaccgagta caagcccacg
    12961 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
    13021 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
    13081 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
    13141 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
    13201 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
    13261 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
    13321 gtcggagtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
    13381 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
    13441 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
    13501 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa cgcgttaagt cgaattcgat
    13561 atcaagctta tcggtaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
    13621 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
    13681 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
    13741 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
    13801 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
    13861 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
    13921 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
    13981 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
    14041 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
    14101 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
    14161 ccgcatcgat accgtcgacc tcgagaccta gaaaaacatg gagcaatcac aagtagcaat
    14221 acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt
    14281 tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
    14341 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
    14401 gatatccttg atctgtggat ctaccacaca caaggctact tccctgattg gcagaactac
    14461 acaccagggc cagggatcag atatccactg acctttggat ggtgctacaa gctagtacca
    14521 gttgagcaag agaaggtaga agaagccaat gaaggagaga acacccgctt gttacaccct
    14581 gtgagcctgc atgggatgga tgacccggag agagaagtat tagagtggag gtttgacagc
    14641 cgcctagcat ttcatcacat ggcccgagag ctgcatccgg actgtactgg gtctctctgg
    14701 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
    14761 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
    14821 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag ggcccgttta
    14881 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
    14941 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
    15001 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
    15061 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
    15121 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
    15181 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
    15241 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
    15301 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
    15361 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
    15421 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
    15481 ccaaactgga acaacactca accctatctc ggtctattct tttg
//
