LOCUS       sequence-410949-       14595 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses FLAG-tagged SpCas9 fused to the N-terminus of Gag/GagPol 
            harbouring an intervening phospholipase C-delta-1 pleckstrin 
            homology (PH) domain .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14595)
  AUTHORS   Haldrup J, Andersen S, Labial ARL, Wolff JH, Frandsen FP, Skov TW, 
            Rovsing AB, Nielsen I, Jakobsen TS, Askou AL, Thomsen MK, Corydon 
            TJ, Thomsen EA, Mikkelsen JG
  TITLE     Engineered lentivirus-derived nanoparticles (LVNPs) for delivery of 
            CRISPR/Cas ribonucleoprotein complexes supporting base editing, 
            prime editing and in vivo gene modification.
  JOURNAL   Nucleic Acids Res. 2023 Sep 8:gkad676. doi: 10.1093/nar/gkad676.
  PUBMED    37678882
REFERENCE   2  (bases 1 to 14595)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2023 Sep 8:gkad676. doi: 10.1093/nar/gkad676."
COMMENT     Sequence Label: pLVNP2.0-SpCas9
FEATURES             Location/Qualifiers
     source          1..14595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             865..930
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             937..957
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             982..5082
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             982..5082
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5083..5130
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     misc_feature    5170..5226
                     /label=MCS
                     /note="multiple cloning site"
     promoter        complement(10149..10167)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(10150..10167)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(10187..10204)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10193..10417
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10463..10890
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10550..10569)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10759..10780
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10899..10919)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10904..11233
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11084..11219
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             11300..12094
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(11354..11373)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     11964..11983
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    12268..12389
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12305..12324)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12359..12378
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12438..12454)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12438..12454)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12451..12473)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12462..12478
                     /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(12486..12492,12493..12510,12511..12516))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12531..12552
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12669..12686)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12840..13428)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12920..12939)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13599..14390,14391..14459))
                     /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     14222..14241
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14460..14564)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
      841 gctagcgcta ccggtaccac catggactat aaggaccacg acggagacta caaggatcat
      901 gatattgatt acaaagacga tgacgataag atggccccaa agaagaagcg gaaggtcggt
      961 atccacggag tcccagcagc cgacaagaag tacagcatcg gcctggacat cggcaccaac
     1021 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg
     1081 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac
     1141 agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga
     1201 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac
     1261 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag
     1321 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc
     1381 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc
     1441 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg
     1501 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag
     1561 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc
     1621 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag
     1681 aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc
     1741 aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac
     1801 ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag
     1861 aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag
     1921 gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg
     1981 ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag
     2041 agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag
     2101 ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac
     2161 agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc
     2221 cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag
     2281 gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct
     2341 ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc
     2401 ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg
     2461 atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg
     2521 tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg
     2581 agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag
     2641 accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc
     2701 ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac
     2761 cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac
     2821 attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa
     2881 cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg
     2941 agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag
     3001 tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg
     3061 cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc
     3121 ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag
     3181 aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac
     3241 aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag
     3301 aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag
     3361 atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac
     3421 tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc
     3481 gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac
     3541 aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag
     3601 gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag
     3661 agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc
     3721 ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc
     3781 ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa
     3841 gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa
     3901 gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga
     3961 accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag
     4021 gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc
     4081 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac
     4141 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg
     4201 gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc
     4261 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg
     4321 aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc
     4381 gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc
     4441 aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc
     4501 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg
     4561 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg
     4621 gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac
     4681 ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag
     4741 aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc
     4801 gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac
     4861 aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc
     4921 ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag
     4981 aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc
     5041 ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg
     5101 aaaaaggccg gccaggcaaa aaagaaaaag aactttagcc agaactatcc gattgtgcag
     5161 accggactca gatctcgagc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg
     5221 ggatccatgg actcgggccg ggacttcctg accctgcacg gcctacagga tgatgaggat
     5281 ctacaggcgc tgctgaaggg cagccagctc ctgaaggtga agtccagctc atggaggaga
     5341 gagcgcttct acaagttgca ggaggactgc aagaccatct ggcaggagtc ccgcaaggtc
     5401 atgcggaccc cggagtccca gctgttctcc atcgaggaca ttcaggaggt gcgaatgggg
     5461 caccgcacgg agggtctgga gaagttcgcc cgtgatgtgc ccgaggaccg ctgcttctcc
     5521 attgtcttca aggaccagcg caatacacta gacctcatcg ccccatcgcc agctgatgcc
     5581 cagcactggg tgctggggct gcacaagatc atccaccact caggctccat ggaccagcgt
     5641 cagaagctac agcactggat tcactcctgc ttgcgaaaag ctgacaaaaa caaggacaac
     5701 aagatgagct tcaaggagct gcagaacttc ctgaaggagc tcaacatcca gcctcgagcc
     5761 gaattcaccc tggccgccag ggccagcgtg ctgagcggcg gcgagctgga caggtgggag
     5821 aagatcaggc tgaggcccgg cggcaagaag aagtataagc tgaagcacat cgtgtgggcc
     5881 agcagggagc tggagaggtt cgccgtgaac cccggcctgc tggagaccag cgagggctgc
     5941 aggcagatcc tgggccagct gcagcccagc ctgcagaccg gcagcgagga gctgaggagc
     6001 ctgtacaaca ccgtggccac cctgtactgc gtgcaccaga ggatcgagat caaggacacc
     6061 aaggaggccc tggacaagat cgaggaggag cagaacaagt ccaagaagaa ggcccagcag
     6121 gccgccgccg acaccggcca cagcagccag gtgagccaga actaccccat cgtgcagaac
     6181 atccagggcc agatggtgca ccaggccatc agccccagga ccctgaacgc ctgggtgaag
     6241 gtggtggagg agaaggcctt cagccccgag gtgatcccca tgttcagcgc cctgagcgag
     6301 ggagccaccc cccaggacct gaacaccatg ctgaacaccg tgggcggcca ccaggccgcc
     6361 atgcagatgc tgaaggagac catcaacgag gaggccgccg agtgggacag ggtgcacccc
     6421 gtgcacgccg gccccatcgc ccccggccag atgagggagc cccgcggcag cgacatcgcc
     6481 ggcaccacca gcaccctgca ggagcagatc ggctggatga ccaacaaccc ccccatcccc
     6541 gtgggcgaaa tctacaagag gtggatcatc ctgggcctga acaagatcgt gaggatgtac
     6601 agccccacca gcatcctgga tatcaggcag ggccccaaag agcccttcag ggactacgtg
     6661 gacaggttct acaagaccct gcgcgccgag caggccagcc aggaggtgaa gaactggatg
     6721 accgagaccc tgctggtgca gaacgccaac cccgactgca agaccatcct gaaggccctg
     6781 ggacccgccg ccaccctgga ggagatgatg accgcctgcc agggcgtggg cggccccggc
     6841 cacaaggcca gggtgctggc cgaggccatg agccaggtga ccaacaccgc caccatcatg
     6901 atgcagaggg gcaacttcag gaaccagagg aagatggtga agtgcttcaa ctgcggcaag
     6961 gagggccaca ccgccaggaa ctgccgcgcc cccaggaaga agggctgctg gaagtgcggc
     7021 aaggagggcc accagatgaa ggactgcacc gagaggcagg ctaatttttt agggaagatc
     7081 tggccttcct acaagggaag gccagggaat tttcttcaga gcagaccaga gccaacagcc
     7141 ccaccatttc ttcagagcag accagagcca acagccccac cagaagagag cttcaggtct
     7201 ggggtagaga caacaactcc ccctcagaag caggagccga tagacaagga actgtatcct
     7261 ttaacttccc tcagatcact ctttggcaac gacccctcgt cacaataaag atcggtggcc
     7321 agctgaagga ggccctgctg gacaccggcg ccgacgacac cgtgctggag gagatgagcc
     7381 tgcccggcag gtggaagccc aagatgatcg gcggcatcgg cggcttcatc aaggtgaggc
     7441 agtacgacca gatcctgatc gagatctgcg gccacaaggc catcggcacc gtgctggtgg
     7501 gacctacacc tgtgaacatc atcggcagga acctgctgac ccagatcggc tgcaccctga
     7561 acttccccat cagccccatc gagaccgtgc ccgtgaagct gaagcccggc atggacggcc
     7621 ctaaggtgaa gcagtggccc ctgaccgagg agaagatcaa ggccctggtg gagatctgca
     7681 ccgagatgga gaaggagggc aagatcagca agatcggccc cgagaacccc tacaacaccc
     7741 ccgtgttcgc catcaagaag aaggacagca ccaagtggag gaagctggtg gacttcaggg
     7801 agctgaacaa gaggacccag gacttctggg aggtgcagct gggcatcccc caccccgccg
     7861 gcctgaagaa gaagaagagc gtgaccgtgc tggacgtggg cgacgcctac ttcagcgtgc
     7921 ccctggacga ggacttcagg aagtataccg ccttcaccat ccccagcatc aacaacgaga
     7981 cccccggcat ccgctaccag tacaacgtgc tgccccaggg ctggaagggc agccccgcca
     8041 tcttccagag cagcatgaca aagatcctgg agcccttcaa gaagcagaac cccgacatcg
     8101 tgatctatca gtacatggac gacctgtacg tgggcagcga cctggagatc ggccagcaca
     8161 ggaccaagat cgaggagctg aggcagcacc tgctgaggtg gggcctgacc acccccgaca
     8221 agaagcacca gaaggagccc ccattcctgt ggatgggcta cgagctgcac cccgacaagt
     8281 ggaccgtgca gcccatcgtg ctgcccgaga aggacagctg gaccgtgaac gacattcaga
     8341 agctggtggg caagctgaac tgggccagcc agatctaccc tggcatcaag gtgaggcagc
     8401 tgtgcaagct gctgaggggc acaaaggctc tgaccgaggt gatccccctg accgaggagg
     8461 ccgagctgga gctggccgag aacagggaga tcctgaagga gcccgtgcac ggcgtgtact
     8521 acgaccccag caaggacctg atcgccgaga tccagaagca gggccagggc cagtggacct
     8581 accagatcta ccaggagccc ttcaagaacc tgaagaccgg caagtacgcc cgcatgcgcg
     8641 gcgcccacac caacgacgtg aagcagctga ccgaggccgt gcagaagatc accaccgaga
     8701 gcatcgtgat ctggggcaag actcctaagt tcaagctgcc catccagaag gagacctggg
     8761 agacctggtg gaccgagtac tggcaggcca cctggattcc cgagtgggag ttcgtgaaca
     8821 cccctcccct ggtgaagctg tggtatcagc tggagaagga gcccatcgtg ggcgccgaga
     8881 ccttctacgt ggacggcgcc gccaacaggg agaccaagct gggcaaggcc ggctacgtga
     8941 ccaacaaggg ccgccagaag gtggtgcccc tgaccaacac caccaaccag aagaccgagc
     9001 tgcaggctat ctacctggcc ctgcaggact caggcctgga ggtgaacatc gtgaccgaca
     9061 gccagtacgc cctgggcatc atccaggccc agcccgacaa gagcgagagc gagctggtga
     9121 accagatcat cgagcagctg atcaagaagg agaaggtgta cctggcctgg gtgcccgccc
     9181 acaagggcat cggcggcaac gagcaggtgg acaagctggt gagcgccggc atcaggaaga
     9241 tcctgttcct ggacggcatc gacaaggccc aggacgagca cgagaagtac cacagcaact
     9301 ggagggctat ggctagcgac ttcaacctgc ctcccgtggt ggctaaggag atcgtggcca
     9361 gctgcgacaa gtgccagctg aagggcgagg ccatgcacgg ccaggtggac tgcagccccg
     9421 gcatctggca gctggtttgc acccacctgg agggcaaggt gatcctggtg gccgtgcacg
     9481 tggcctccgg ctacatcgag gccgaggtga tccccgccga gaccggccag gagaccgcct
     9541 acttcctgct gaagctggcc ggccgctggc ccgtgaagac catccacacc gacaacggca
     9601 gcaacttcac cagcgccacc gtgaaggctg cctgctggtg ggccggcatc aagcaggagt
     9661 tcggcatccc ctacaacccc cagtctcagg gcgtggtgga gagcatgaac aaggagctga
     9721 agaagatcat cggccaggtg agggaccagg ccgagcacct gaagaccgcc gtgcagatgg
     9781 ccgtgttcat ccacaacttc aagaggaagg gcggcatcgg cggctacagc gccggcgaga
     9841 ggatcgtgga catcatcgcc accgacatcc agaccaagga gctgcagaag cagatcacca
     9901 agatccagaa cttcagggtg tactacaggg acagcaggaa ccctctgtgg aagggccccg
     9961 ccaagctgct gtggaagggc gagggcgccg tggtgatcca ggacaacagc gacatcaagg
    10021 tggtgcccag gaggaaggcc aagatcatca gggactacgg caagcagatg gccggcgacg
    10081 actgcgtggc ctccaggcag gacgaggact gataaggatc cctcgagcat gcatctagag
    10141 ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct
    10201 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
    10261 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
    10321 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
    10381 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
    10441 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
    10501 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
    10561 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aatcgggggc
    10621 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
    10681 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
    10741 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
    10801 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
    10861 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
    10921 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
    10981 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
    11041 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
    11101 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
    11161 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
    11221 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
    11281 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
    11341 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
    11401 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
    11461 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
    11521 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
    11581 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
    11641 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
    11701 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
    11761 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
    11821 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
    11881 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
    11941 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
    12001 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
    12061 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
    12121 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
    12181 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
    12241 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
    12301 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
    12361 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
    12421 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
    12481 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
    12541 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
    12601 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
    12661 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
    12721 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
    12781 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
    12841 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
    12901 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
    12961 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
    13021 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
    13081 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
    13141 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
    13201 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
    13261 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
    13321 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
    13381 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
    13441 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
    13501 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
    13561 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
    13621 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
    13681 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
    13741 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
    13801 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
    13861 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
    13921 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
    13981 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
    14041 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
    14101 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
    14161 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
    14221 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
    14281 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
    14341 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
    14401 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
    14461 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
    14521 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
    14581 gtgccacctg acgtc
//
