LOCUS       sequence-252041-       13451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral expression of SpCas9 under hUBC promotor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13451)
  AUTHORS   Willems J, de Jong APH, Scheefhals N, Mertens E, Catsburg LAE, 
            Poorthuis RB, de Winter F, Verhaagen J, Meye FJ, MacGillavry HD
  TITLE     ORANGE: A CRISPR/Cas9-based genome editing toolbox for epitope 
            tagging of endogenous proteins in neurons.
  JOURNAL   PLoS Biol. 2020 Apr 10;18(4):e3000665. doi: 
            10.1371/journal.pbio.3000665.
  PUBMED    32275651
REFERENCE   2  (bases 1 to 13451)
  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: "PLoS 
            Biol."; date: "2020-04-10"; volume: "18(4):e3000665. doi"; pages: " 
            10.1371/journal.pbio.3000665"
COMMENT     Sequence Label: pFUGW SpCas9
FEATURES             Location/Qualifiers
     source          1..13451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(56..75)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        247..626
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        628..826
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     781..801
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             844..1024
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1071..1196
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1689..1922
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2107..2151
                     /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    2449..2566
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2631..3030
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3678..3697
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3922..3948
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3952..3972
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3982..8082
                     /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             3982..8082
                     /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             8086..8106
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    8143..8731
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8196..8216)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8614..8625)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8734..8750)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8735..8751)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9256..9436
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9462..9479)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9468..9692
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9738..10166
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9825..9844)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10035..10056
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10175..10195)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10180..10509
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10360..10495
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10422..10441
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10557..10604
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10623..10997
                     /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    11127..11248
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11164..11183)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11218..11237
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11297..11313)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11297..11313)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11310..11332)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11321..11337
                     /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(11345..11351,11352..11369,11370..11375))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11390..11411
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11528..11545)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11699..12287)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11779..11798)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12458..13249,13250..13318))
                     /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     13081..13100
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13319..13423)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 attnnntncg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
       61 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
      121 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
      181 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      241 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      601 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
      841 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
      901 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
      961 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     1021 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     1081 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     1141 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     1201 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     1261 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     1321 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     1381 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     1441 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     1501 agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga
     1561 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     1621 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     1681 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     1741 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     1801 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     1861 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     1921 ctggggattt ggggttgctc tggaaaattc atttgcacca ctgctgtgcc ttggaatgct
     1981 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     2041 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     2101 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     2161 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     2221 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     2281 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     2341 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg
     2401 gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa
     2461 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     2521 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     2581 ggacagcaga gatccagttt ggttaattaa cccgtgtcgg ctccagatct ggcctccgcg
     2641 ccgggttttg gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg ccacgtcaga
     2701 cgaagggcgc aggagcgttc ctgatccttc cgcccggacg ctcaggacag cggcccgctg
     2761 ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt
     2821 gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc
     2881 ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa
     2941 ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt
     3001 gtttgtggat cgctgtgatc gtcacttggt gagttgcggg ctgctgggct ggccggggct
     3061 ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt
     3121 agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg cacaaaatgg
     3181 cggctgttcc cgagtcttga atggaagacg cttgtaaggc gggctgtgag gtcgttgaaa
     3241 caaggtgggg ggcatggtgg gcggcaagaa cccaaggtct tgaggccttc gctaatgcgg
     3301 gaaagctctt attcgggtga gatgggctgg ggcaccatct ggggaccctg acgtgaagtt
     3361 tgtcactgac tggagaactc gggtttgtcg tctggttgcg ggggcggcag ttatgcggtg
     3421 ccgttgggca gtgcacccgt acctttggga gcgcgcgcct cgtcgtgtcg tgacgtcacc
     3481 cgttctgttg gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt
     3541 tctccgtcgc aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc
     3601 ggacctctgg tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac
     3661 ctatcttctt aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt
     3721 gttttgtgaa gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt
     3781 tcagtgttag actagtaaag cttctgcagg tcgactctag aaaattgtcc gctaaattct
     3841 ggccgttttt ggcttttttg ttagacgaag cttgggctgc aggtcgactc tagaggatcc
     3901 ccgggtaccg gtgccaccat gtacccatac gatgttccag attacgcttc gccgaagaaa
     3961 aagcgcaagg tcgaagcgtc cgacaagaag tacagcatcg gcctggacat cggcaccaac
     4021 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg
     4081 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac
     4141 agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga
     4201 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac
     4261 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag
     4321 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc
     4381 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc
     4441 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg
     4501 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag
     4561 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc
     4621 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag
     4681 aatggcctgt tcggcaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc
     4741 aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac
     4801 ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag
     4861 aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag
     4921 gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg
     4981 ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag
     5041 agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag
     5101 ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac
     5161 agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc
     5221 cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag
     5281 gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct
     5341 ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc
     5401 ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg
     5461 atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg
     5521 tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg
     5581 agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag
     5641 accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc
     5701 ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac
     5761 cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac
     5821 attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa
     5881 cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg
     5941 agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag
     6001 tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg
     6061 cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc
     6121 ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag
     6181 aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac
     6241 aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag
     6301 aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag
     6361 atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac
     6421 tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc
     6481 gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac
     6541 aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag
     6601 gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag
     6661 agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc
     6721 ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc
     6781 ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa
     6841 gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa
     6901 gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga
     6961 accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag
     7021 gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc
     7081 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac
     7141 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg
     7201 gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc
     7261 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg
     7321 aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc
     7381 gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc
     7441 aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc
     7501 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg
     7561 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg
     7621 gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac
     7681 ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag
     7741 aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc
     7801 gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac
     7861 aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc
     7921 ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag
     7981 aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc
     8041 ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acagccccaa gaagaagaga
     8101 aaggtggagg ccagctaaga attcgatatc aagcttatcg ataatcaacc tctggattac
     8161 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     8221 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     8281 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     8341 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     8401 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     8461 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     8521 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     8581 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     8641 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     8701 agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg agacctagaa
     8761 aaacatggag caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta
     8821 gaagcacaag aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca
     8881 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     8941 gggctaattc actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa
     9001 ggctacttcc ctgattggca gaactacaca ccagggccag ggatcagata tccactgacc
     9061 tttggatggt gctacaagct agtaccagtt gagcaagaga aggtagaaga agccaatgaa
     9121 ggagagaaca cccgcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga
     9181 gaagtattag agtggaggtt tgacagccgc ctagcatttc atcacatggc ccgagagctg
     9241 catccggact gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     9301 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     9361 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     9421 tggaaaatct ctagcagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     9481 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     9541 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     9601 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     9661 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     9721 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     9781 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     9841 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     9901 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     9961 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
    10021 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
    10081 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
    10141 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
    10201 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
    10261 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
    10321 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
    10381 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
    10441 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
    10501 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg
    10561 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
    10621 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
    10681 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
    10741 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
    10801 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
    10861 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
    10921 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
    10981 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
    11041 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
    11101 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
    11161 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
    11221 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
    11281 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
    11341 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
    11401 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
    11461 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
    11521 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
    11581 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
    11641 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
    11701 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
    11761 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
    11821 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
    11881 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
    11941 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
    12001 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
    12061 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
    12121 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
    12181 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
    12241 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
    12301 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
    12361 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
    12421 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
    12481 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
    12541 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
    12601 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
    12661 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
    12721 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
    12781 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
    12841 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
    12901 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
    12961 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
    13021 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
    13081 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
    13141 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
    13201 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
    13261 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
    13321 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
    13381 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
    13441 tgccacctga c
//
