LOCUS       sequence-450926-       15072 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Variant of lentiCRISPRv2 that confers G418 resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15072)
  AUTHORS   Stringer BW, Day BW, D'Souza RCJ, Jamieson PR, Ensbey KS, Bruce ZC, 
            Lim YC, Goasdoue K, Offenhauser C, Akgul S, Allan S, Robertson T, 
            Lucas P, Tollesson G, Campbell S, Winter C, Do H, Dobrovic A, Inglis
            PL, Jeffree RL, Johns TG, Boyd AW
  TITLE     A reference collection of patient-derived cell line and xenograft 
            models of proneural, classical and mesenchymal glioblastoma.
  JOURNAL   Sci Rep. 2019 Mar 20;9(1):4902. doi: 10.1038/s41598-019-41277-z.
  PUBMED    30894629
REFERENCE   2  (bases 1 to 15072)
  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: "Sci Rep."; 
            date: "2019-03-20"; volume: "9(1):4902. doi"; pages: " 
            10.1038/s41598-019-41277-z"
COMMENT     Sequence Label: lentiCRISPRv2 neo
FEATURES             Location/Qualifiers
     source          1..15072
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1040..1161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1051..1070)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1105..1124
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1291..1665)
                     /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"
     promoter        complement(1684..1731)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(1779..2108)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1793..1928
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1847..1866)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2093..2113
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2122..2550)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2232..2253)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2444..2463
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2596..2820
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2809..2826
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             2852..3085
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    3157..3745
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3263..3274
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3672..3692
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3761..4552)
                     /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="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3872..3891)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4482..4501
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(4553..4609)
                     /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             complement(4619..4642)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(4643..4690)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(4691..8794)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(4691..8794)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     regulatory      8791..8800
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(8819..9030)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_RNA        complement(9092..9167)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(11049..11297)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(11107..11126)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(11277..11297)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    11348..11465
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(11763..11807)
                     /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    11992..12225
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    12718..12843
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             12890..13070
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(13088..13286)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(13113..13133)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        13288..13667
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     13843..13862
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        13937..14041
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(14042..14110,14111..14902)
                     /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     complement(14260..14279)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
     1021 cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1081 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1141 ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg
     1201 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
     1261 ggcggtggaa tcgaaatctc gtagcacgtg tcagtcctgc tcctcggcca cgaagtgcac
     1321 gcagttgccg gccgggtcgc gcagggcgaa ctcccgcccc cacggctgct cgccgatctc
     1381 ggtcatggcc ggcccggagg cgtcccggaa gttcgtggac acgacctccg accactcggc
     1441 gtacagctcg tccaggccgc gcacccacac ccaggccagg gtgttgtccg gcaccacctg
     1501 gtcctggacc gcgctgatga acagggtcac gtcgtcccgg accacaccgg cgaagtcgtc
     1561 ctccacgaag tcccgggaga acccgagccg gtcggtccag aactcgaccg ctccggcgac
     1621 gtcgcgcgcg gtgagcaccg gaacggcact ggtcaacttg gccatggttt agttcctcac
     1681 cttgtcgtat tatactatgc cgatatacta tgccgatgat taattgtcaa cacgtgctga
     1741 tcagatccga aaatggatat acaagctccc gggagctttt tgcaaaagcc taggcctcca
     1801 aaaaagcctc ctcactactt ctggaatagc tcagaggcag aggcggcctc ggcctctgca
     1861 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
     1921 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
     1981 tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg
     2041 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
     2101 tgacacacat tccacagaat taattcgcgt taaatttttg ttaaatcagc tcatttttta
     2161 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     2221 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     2281 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     2341 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     2401 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     2461 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     2521 ccgcgcttaa tgcgccgcta cagggcgcgt ggggataccc cctagagccc cagctggttc
     2581 tttccgcctc agaagccata gagcccaccg catccccagc atgcctgcta ttgtcttccc
     2641 aatcctcccc cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca
     2701 gacaatgcga tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca
     2761 gggtcaagga aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag
     2821 tcgaggctga tcagcgggtt taaacgggcc ctgctagaga ttttccacac tgactaaaag
     2881 ggtctgaggg atctctagtt accagagtca cacaacagac gggcacacac tacttgaagc
     2941 actcaaggca agctttattg aggcttaagc agtgggttcc ctagttagcc agagagctcc
     3001 caggctcaga tctggtctaa ccagagagac ccagtacaag caaaaagcag atcttgtctt
     3061 cgttgggagt gaattagccc ttccagtccc cccttttctt ttaaaaagtg gctaagatct
     3121 acagctgcct tgtaagtcat tggtcttaaa gtcgacgcgg ggaggcggcc caaagggaga
     3181 tccgactcgt ctgagggcga aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc
     3241 cgcgggaagg aaggtccgct ggattgaggg ccgaagggac gtagcagaag gacgtcccgc
     3301 gcagaatcca ggtggcaaca caggcgagca gccaaggaaa ggacgatgat ttccccgaca
     3361 acaccacgga attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg
     3421 cggcgatgag ttccgccgtg gcaataggga gggggaaagc gaaagtcccg gaaaggagct
     3481 gacaggtggt ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca gtgcacacca
     3541 cgccacgttg cctgacaacg ggccacaact cctcataaag agacagcaac caggatttat
     3601 acaaggagga gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag
     3661 cagcgtatcc acatagcgta aaaggagcaa catagttaag aataccagtc aatctttcac
     3721 aaattttgta atccagaggt tgattgtcga cttaacgcgt tcagaagaac tcgtcaagaa
     3781 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
     3841 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
     3901 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
     3961 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg
     4021 gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
     4081 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat
     4141 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
     4201 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc
     4261 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag
     4321 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt
     4381 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
     4441 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
     4501 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcggtccag
     4561 gattctcttc gacatctccg gcttgtttca gcagagagaa gtttgttgcg ccggatccct
     4621 tatcgtcatc gtctttgtaa tctttcttct tcttagcctg tccagccttc tttgtggcgg
     4681 caggtcgctt gtcgcctccc agctgagaca ggtcgatccg tgtctcgtac aggccggtga
     4741 tgctctggtg gatcagggtg gcgtccagca cctctttggt gctggtgtac ctcttccggt
     4801 cgatggtggt gtcaaagtac ttgaaggcgg caggggctcc cagattggtc agggtaaaca
     4861 ggtggatgat attctcggcc tgctctctga tgggcttatc ccggtgcttg ttgtaggcgg
     4921 acagcacttt gtccagatta gcgtcggcca ggatcactct cttggagaac tcgctgatct
     4981 gctcgatgat ctcgtccagg tagtgcttgt gctgttccac aaacagctgt ttctgctcat
     5041 tatcctcggg ggagcccttc agcttctcat agtggctggc caggtacagg aagttcacat
     5101 atttggaggg cagggccagt tcgtttccct tctgcagttc gccggcagag gccagcattc
     5161 tcttccggcc gttttccagc tcgaacaggg agtacttagg cagcttgatg atcaggtcct
     5221 ttttcacttc tttgtagccc ttggcttcca gaaagtcgat gggattcttc tcgaagctgc
     5281 ttctttccat gatggtgatc cccagcagct ctttcacact cttcagtttc ttggacttgc
     5341 ccttttccac tttggccacc accagcacag aataggccac ggtggggctg tcgaagccgc
     5401 cgtacttctt agggtcccag tccttctttc tggcgatcag cttatcgctg ttcctcttgg
     5461 gcaggataga ctctttgctg aagccgcctg tctgcacctc ggtctttttc acgatattca
     5521 cttggggcat gctcagcact ttccgcacgg tggcaaaatc ccggccctta tcccacacga
     5581 tctccccggt ttcgccgttt gtctcgatca gaggccgctt ccggatctcg ccgttggcca
     5641 gggtaatctc ggtcttgaaa aagttcatga tgttgctgta gaagaagtac ttggcggtag
     5701 ccttgccgat ttcctgctcg ctcttggcga tcatcttccg cacgtcgtac accttgtagt
     5761 cgccgtacac gaactcgctt tccagcttag ggtacttttt gatcagggcg gttcccacga
     5821 cggcgttcag gtaggcgtcg tgggcgtggt ggtagttgtt gatctcgcgc actttgtaaa
     5881 actggaaatc cttccggaaa tcggacacca gcttggactt cagggtgatc actttcactt
     5941 cccggatcag cttgtcattc tcgtcgtact tagtgttcat ccgggagtcc aggatctgtg
     6001 ccacgtgctt tgtgatctgc cgggtttcca ccagctgtct cttgatgaag ccggccttat
     6061 ccagttcgct caggccgcct ctctcggcct tggtcagatt gtcgaacttt ctctgggtaa
     6121 tcagcttggc gttcagcagc tgccgccagt agttcttcat cttcttcacg acctcttcgg
     6181 agggcacgtt gtcgctcttg ccccggttct tgtcgcttct ggtcagcacc ttgttgtcga
     6241 tggagtcgtc cttcagaaag ctctgaggca cgatatggtc cacatcgtag tcggacagcc
     6301 ggttgatgtc cagttcctgg tccacgtaca tatcccgccc attctgcagg tagtacaggt
     6361 acagcttctc gttctgcagc tgggtgtttt ccacggggtg ttctttcagg atctggctgc
     6421 ccagctcttt gatgccctct tcgatccgct tcattctctc gcggctgttc ttctgtccct
     6481 tctgggtggt ctggttctct ctggccattt cgatcacgat gttctcgggc ttgtgccggc
     6541 ccatcacttt cacgagctcg tccaccacct tcactgtctg caggatgccc ttcttaatgg
     6601 cggggctgcc ggccagattg gcaatgtgct cgtgcaggct atcgccctgg ccggacacct
     6661 gggctttctg gatgtcctct ttaaaggtca ggctgtcgtc gtggatcagc tgcatgaagt
     6721 ttctgttggc gaagccgtcg gacttcagga aatccaggat tgtcttgccg gactgcttgt
     6781 cccggatgcc gttgatcagc ttccggctca gcctgcccca gccggtgtat ctccgccgct
     6841 tcagctgctt catcactttg tcgtcgaaca ggtgggcata ggttttcagc cgttcctcga
     6901 tcatctctct gtcctcaaac agtgtcaggg tcagcacgat atcttccaga atgtcctcgt
     6961 tttcctcatt gtccaggaag tccttgtcct tgataatttt cagcagatcg tggtatgtgc
     7021 ccagggaggc gttgaaccga tcttccacgc cggagatttc cacggagtcg aagcactcga
     7081 ttttcttgaa gtagtcctct ttcagctgct tcacggtcac tttccggttg gtcttgaaca
     7141 gcaggtccac gatggccttt ttctgctcgc cgctcaggaa ggcgggcttt ctcattccct
     7201 cggtcacgta tttcactttg gtcagctcgt tatacacggt gaagtactcg tacagcaggc
     7261 tgtgcttggg cagcaccttc tcgttgggca ggttcttatc gaagttggtc atccgctcga
     7321 tgaagctctg ggcggaagcg cccttgtcca ccacttcctc gaagttccag ggggtgatgg
     7381 tttcctcgct ctttctggtc atccaggcga atctgctgtt tcccctggcc agagggccca
     7441 cgtagtaggg gatgcggaag gtcaggatct tctcgatctt ttcccggttg tccttcagga
     7501 atgggtaaaa atcttcctgc cgccgcagaa tggcgtgcag ctctcccagg tggatctggt
     7561 gggggatgct gccgttgtcg aaggtccgct gcttccgcag caggtcctct ctgttcagct
     7621 tcacgagcag ttcctcggtg ccgtccatct tttccaggat gggcttgatg aacttgtaga
     7681 actcttcctg gctggctccg ccgtcaatgt agccggcgta gccgttcttg ctctggtcga
     7741 agaaaatctc tttgtacttc tcaggcagct gctgccgcac gagagctttc agcagggtca
     7801 ggtcctggtg gtgctcgtcg tatctcttga tcatagaggc gctcaggggg gccttggtga
     7861 tctcggtgtt cactctcagg atgtcgctca gcaggatggc gtcggacagg ttcttggcgg
     7921 ccagaaacag gtcggcgtac tggtcgccga tctgggccag caggttgtcc aggtcgtcgt
     7981 cgtaggtgtc cttgctcagc tgcagtttgg catcctcggc caggtcgaag ttgctcttga
     8041 agttgggggt caggcccagg ctcagggcaa tcaggtttcc gaacaggcca ttcttcttct
     8101 cgccgggcag ctgggcgatc agattttcca gccgtctgct cttgctcagt ctggcagaca
     8161 ggatggcctt ggcgtccacg ccgctggcgt tgatggggtt ttcctcgaac agctggttgt
     8221 aggtctgcac cagctggatg aacagcttgt ccacgtcgct gttgtcgggg ttcaggtcgc
     8281 cctcgatcag gaagtggccc cggaacttga tcatgtgggc cagggccaga tagatcagcc
     8341 gcaggtcggc cttgtcggtg ctgtccacca gtttctttct caggtggtag atggtggggt
     8401 acttctcgtg gtaggccacc tcgtccacga tgttgccgaa gatggggtgc cgctcgtgct
     8461 tcttatcctc ttccaccagg aaggactctt ccagtctgtg gaagaagctg tcgtccacct
     8521 tggccatctc gttgctgaag atctcttgca gatagcagat ccggttcttc cgtctggtgt
     8581 atcttcttct ggcggttctc ttcagccggg tggcctcggc tgtttcgccg ctgtcgaaca
     8641 gcagggctcc gatcaggttc ttcttgatgc tgtgccggtc ggtgttgccc agcaccttga
     8701 atttcttgct gggcaccttg tactcgtcgg tgatcacggc ccagcccaca gagttggtgc
     8761 cgatgtccag gccgatgctg tacttcttgt ccatggtggc agcgctctag aaccggtcct
     8821 gtgttctggc ggcaaacccg ttgcgaaaaa gaacgttcac ggcgactact gcacttatat
     8881 acggttctcc cccaccctcg ggaaaaaggc ggagccagta cacgacatca ctttcccagt
     8941 ttaccccgcg ccaccttctc taggcaccgg atcaattgcc gacccctccc cccaacttct
     9001 cggggactgt gggcgatgtg cgctctgccc actgacgggc accggagcca attcccactc
     9061 ctttcaagac ctagctagcg aattcaaaaa agcaccgact cggtgccact ttttcaagtt
     9121 gataacggac tagccttatt ttaacttgct atttctagct ctaaaacaga gacgtacaaa
     9181 aaagagcaag aagctaaaaa agatttaaaa attattttta gcgcagttaa tggaacagga
     9241 actaaattta ccccaaaaat attacgtgaa tcaggatata acgttattga ggttgaagag
     9301 catgcatttg aagatgaaac atttaaaaat gttgtaaatc caaatccaga atttgatcct
     9361 gcatgaaaaa taccgcttga atatggtatt aaacatgatg cagatattat tattatgaat
     9421 gacccagatg ctgacagatt tggaatggca ataaaacatg atggtcattt tgtaagatta
     9481 gatggaaatc aaacaggacc aattttaatt gattgaaaat tatcaaatct aaaacgctta
     9541 aatagcattc caaaaaatcc ggctctatat tcaagttttg taacaagtga tttgggtgat
     9601 agaatcgctc atgaaaaata tggagttaat attgtaaaaa ctttaactgg atttaaatga
     9661 atgggtagag aaattgctaa agaagaagat aacggattaa attttgtttt tgcttatgaa
     9721 gaaagttatg gatatgtaat tgatgactca gctagagata aagatggaat acaagcttct
     9781 atattaatag cagaggctgc ttgattttat aaaaaacaaa ataaaacatt agtagactat
     9841 ttagaagatt tatttaaaga aatgggtgca tattacactt tcactttaaa cttgaatttt
     9901 aaaccagaag aaaagaaatt aaaaattgaa ccattaatga aatcattgag agcaacaccc
     9961 ttaactcaaa ttgctggact taaagttgtt aatgttgaag actacatcga tggaatgtat
    10021 aatatgccag gacaagactt actaaaattt tatttagaag ataagtcatg atttgctgtt
    10081 cgcccaagtg gaactgaacc taaactaaaa atttatttta taggtgttgg tgaatctgtt
    10141 caaaacgcta aagttaaagt agacgaaatt attaaagaat taaaattaaa aatgaatata
    10201 taggagaaaa aatgaaacta aacaaatata tagatcacac attattaaaa caagatgcta
    10261 cgaaagctga aattaaacaa ttatgtgatg aagcaattga atttgatttt gcaacagttt
    10321 gtgttaattc atattgaaca agctattgta aagaattatt aaaaggcaca aatgtaggaa
    10381 taacaaatgt tgtaggtttt cctctaggtg catgcacaac agctacaaaa gcattcgaag
    10441 tttctgaagc aattaaagat ggtgcaacag aaattgatat ggtattaaat attggtgcat
    10501 taaaagacaa aaattatgaa ttagttttag aagacatgaa agctgtaaaa aaagcagctg
    10561 gatcacatgt tgttaaatgt attatggaaa attgtttatt aacaaaagaa gaaatcatga
    10621 aagcttgtga aatagctgtt gaagctggat tagaatttgt taaaacatca acaggatttt
    10681 caaaatcagg tgcaacattt gaagatgtta aactaatgaa gtcagttgtt aaagacaatg
    10741 ctttagttaa agcagctggt ggagttagaa catttgaaga tgctcaaaaa atgattgaag
    10801 caggagctga ccgcttagga acaagtggtg gagtagctat tattaaaggt gaagaaaaca
    10861 acgcgagtta ctaaaactag cgttttttta ttttgctcat ttttattaaa agtttgcaaa
    10921 aaggaacata aaaattctaa ttattgatac taaagttatt aaaaagaaga ttttggttga
    10981 ttttataaag gtcatagaat ataatatttt agcatgtgta ttttgtgtgc tcatttacaa
    11041 ccgtctccgg tgtttcgtcc tttccacaag atatataaag ccaagaaatc gaaatacttt
    11101 caagttacgg taagcatatg atagtccatt ttaaaacata attttaaaac tgcaaactac
    11161 ccaagaaatt attactttct acgtcacgta ttttgtacta atatctttgt gtttacagtc
    11221 aaattaattc taattatctc tctaacagcc ttgtatcgta tatgcaaata tgaaggaatc
    11281 atgggaaata ggccctcggt accttaatta accaaactgg atctctgctg tccctgtaat
    11341 aaacccgaaa attttgaatt tttgtaattt gtttttgtaa ttctttagtt tgtatgtctg
    11401 ttgctattat gtctactatt ctttcccctg cactgtaccc cccaatcccc ccttttcttt
    11461 taaaattgtg gatgaatact gccatttgtc tgcagaattg gcgcacgcag tgccgatccg
    11521 ttcactaatc gaatggatct gtctctgtct ctctctccac cttcttcttc tattccttcg
    11581 ggcctgtcgg gtcccctcgg ggttgggagg tgggtctgaa acgataatgg tgaatatccc
    11641 tgcctaactc tattcactat agaaagtaca gcaaaaacta ttcttaaacc taccaagcct
    11701 cctactatca ttatgaataa ttttatatac cacagccaat ttgttatgtt aaaccaattc
    11761 cacaaacttg cccatttatc taattccaat aattcttgtt cattcttttc ttgctggttt
    11821 tgcgattctt caattaagga gtgtattaag cttgtgtaat tgttaatttc tctgtcccac
    11881 tccatccagg tcgtgtgatt ccaaatctgt tccagagatt tattactcca actagcattc
    11941 caaggcacag cagtggtgca aatgagtttt ccagagcaac cccaaatccc caggagctgt
    12001 tgatccttta ggtatctttc cacagccagg attcttgcct ggagctgctt gatgccccag
    12061 actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt gttctgctgc
    12121 tgcactatac cagacaataa ttgtctggcc tgtaccgtca gcgtcattga cgctgcgccc
    12181 atagtgcttc ctgctgctcc caagaaccca aggaacaaag ctcctattcc cactgctctt
    12241 ttttctctct gcaccactct tctctttgcc ttggtgggtg ctactcctaa tggttcaatt
    12301 tttactactt tatatttata taattcactt ctccaattgt ccctcatatc tcctcctcca
    12361 ggtctgaaga tcagcggccg cttgctgtgc ggtggtctta cttttgtttt gctcttcctc
    12421 tatcttgtct aaagcttcct tggtgtcttt tatctctatc ctttgatgca cacaatagag
    12481 ggttgctact gtattatata atgatctaag ttcttctgat cctgtctgaa gggatggttg
    12541 tagctgtccc agtatttgtc tacagccttc tgatgtttct aacaggccag gattaactgc
    12601 gaatcgttct agctccctgc ttgcccatac tatatgtttt aatttatatt ttttctttcc
    12661 ccctggcctt aaccgaattt tttcccatcg cgatctaatt ctcccccgct taatactgac
    12721 gctctcgcac ccatctctct ccttctagcc tccgctagtc aaaatttttg gcgtactcac
    12781 cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga
    12841 gagctcctct ggtttccctt tcgctttcaa gtccctgttc gggcgccact gctagagatt
    12901 ttccacactg actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg
    12961 gcacacacta cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct
    13021 agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc agtacaggca
    13081 aaacgcgctg cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg
    13141 ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc
    13201 attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc
    13261 attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta
    13321 ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt
    13381 accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa
    13441 gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt
    13501 tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc
    13561 aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa
    13621 tgaccccgta attgattact attaataact agtcaataat caatgtcaac gcgcgcgtat
    13681 atctggcccg tacatcgcga agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc
    13741 aattgtcggt caagccttgc cttgttgtag cttaaatttt gctcgcgcac tactcagcga
    13801 cctccaacac acaagcaggg agcagatact ggcttaacta tgcggcatca gagcagattg
    13861 tactgagagt gcaccatagg ggatcgggag atctcccgat ccgtcgacgt caggtggcac
    13921 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
    13981 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
    14041 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
    14101 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
    14161 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
    14221 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
    14281 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
    14341 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
    14401 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
    14461 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
    14521 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
    14581 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
    14641 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
    14701 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
    14761 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
    14821 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
    14881 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
    14941 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
    15001 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
    15061 caaaggatct tc
//
