LOCUS       Exported               15072 bp ds-DNA     circular SYN 19-JAN-2022
DEFINITION  Variant of lentiCRISPRv2 that confers G418 resistance.
ACCESSION   .
VERSION     .
KEYWORDS    lentiCRISPRv2 neo
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 Jan 19, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..15072
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     39..60
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(179..199)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        184..513
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      364..499
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     426..445
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        561..608
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             627..1001
                     /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    1131..1252
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1168..1187)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1222..1241
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1301..1317)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1301..1317)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1314..1336)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1325..1341
                     /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(1349..1379)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1394..1415
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1532..1549)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1703..2291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1783..1802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2462..3322)
                     /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     3085..3104
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3323..3427)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3502..3521)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3697..4076
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4078..4276
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4231..4251
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4294..4474
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4521..4646
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5139..5372
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5557..5601
                     /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    5899..6016
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6067..6307
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     6067..6087
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6238..6257
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8197..8272
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8334..8545
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      8564..8573
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             8570..12673
                     /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             12674..12721
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             12722..12745
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             12755..12811
                     /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             12812..13603
                     /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(12863..12882)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     13473..13492
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    13619..14207
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(13672..13692)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(14090..14101)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             14279..14512
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(14538..14555)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    14544..14768
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      14814..170
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14901..14920)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
       61 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
      121 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
      181 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
      241 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
      301 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
      361 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
      421 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
      481 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
      541 ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat
      601 aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca
      661 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg
      721 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
      781 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg
      841 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc
      901 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg
      961 ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg
     1021 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     1081 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     1141 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     1201 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     1261 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     1321 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     1381 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     1441 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     1501 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     1561 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1621 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1681 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1741 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1801 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1861 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1921 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1981 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2041 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2101 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2161 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2221 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2281 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2341 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2401 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2461 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2521 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     2581 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     2641 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     2701 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     2761 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     2821 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     2881 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     2941 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3001 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3061 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3121 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3181 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3241 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3301 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3361 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3421 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     3481 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     3541 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     3601 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     3661 gcttcgcgat gtacgggcca gatatacgcg cgcgttgaca ttgattattg actagttatt
     3721 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     3781 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     3841 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     3901 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     3961 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     4021 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     4081 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     4141 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     4201 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     4261 aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga ccagatctga
     4321 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     4381 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     4441 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
     4501 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
     4561 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
     4621 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
     4681 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
     4741 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
     4801 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     4861 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     4921 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     4981 aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
     5041 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
     5101 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
     5161 ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga
     5221 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
     5281 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
     5341 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
     5401 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
     5461 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
     5521 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
     5581 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
     5641 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
     5701 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
     5761 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
     5821 gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat
     5881 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
     5941 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
     6001 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt ggttaattaa
     6061 ggtaccgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     6121 agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     6181 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     6241 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     6301 aaaggacgaa acaccggaga cggttgtaaa tgagcacaca aaatacacat gctaaaatat
     6361 tatattctat gacctttata aaatcaacca aaatcttctt tttaataact ttagtatcaa
     6421 taattagaat ttttatgttc ctttttgcaa acttttaata aaaatgagca aaataaaaaa
     6481 acgctagttt tagtaactcg cgttgttttc ttcaccttta ataatagcta ctccaccact
     6541 tgttcctaag cggtcagctc ctgcttcaat cattttttga gcatcttcaa atgttctaac
     6601 tccaccagct gctttaacta aagcattgtc tttaacaact gacttcatta gtttaacatc
     6661 ttcaaatgtt gcacctgatt ttgaaaatcc tgttgatgtt ttaacaaatt ctaatccagc
     6721 ttcaacagct atttcacaag ctttcatgat ttcttctttt gttaataaac aattttccat
     6781 aatacattta acaacatgtg atccagctgc tttttttaca gctttcatgt cttctaaaac
     6841 taattcataa tttttgtctt ttaatgcacc aatatttaat accatatcaa tttctgttgc
     6901 accatcttta attgcttcag aaacttcgaa tgcttttgta gctgttgtgc atgcacctag
     6961 aggaaaacct acaacatttg ttattcctac atttgtgcct tttaataatt ctttacaata
     7021 gcttgttcaa tatgaattaa cacaaactgt tgcaaaatca aattcaattg cttcatcaca
     7081 taattgttta atttcagctt tcgtagcatc ttgttttaat aatgtgtgat ctatatattt
     7141 gtttagtttc attttttctc ctatatattc atttttaatt ttaattcttt aataatttcg
     7201 tctactttaa ctttagcgtt ttgaacagat tcaccaacac ctataaaata aatttttagt
     7261 ttaggttcag ttccacttgg gcgaacagca aatcatgact tatcttctaa ataaaatttt
     7321 agtaagtctt gtcctggcat attatacatt ccatcgatgt agtcttcaac attaacaact
     7381 ttaagtccag caatttgagt taagggtgtt gctctcaatg atttcattaa tggttcaatt
     7441 tttaatttct tttcttctgg tttaaaattc aagtttaaag tgaaagtgta atatgcaccc
     7501 atttctttaa ataaatcttc taaatagtct actaatgttt tattttgttt tttataaaat
     7561 caagcagcct ctgctattaa tatagaagct tgtattccat ctttatctct agctgagtca
     7621 tcaattacat atccataact ttcttcataa gcaaaaacaa aatttaatcc gttatcttct
     7681 tctttagcaa tttctctacc cattcattta aatccagtta aagtttttac aatattaact
     7741 ccatattttt catgagcgat tctatcaccc aaatcacttg ttacaaaact tgaatataga
     7801 gccggatttt ttggaatgct atttaagcgt tttagatttg ataattttca atcaattaaa
     7861 attggtcctg tttgatttcc atctaatctt acaaaatgac catcatgttt tattgccatt
     7921 ccaaatctgt cagcatctgg gtcattcata ataataatat ctgcatcatg tttaatacca
     7981 tattcaagcg gtatttttca tgcaggatca aattctggat ttggatttac aacattttta
     8041 aatgtttcat cttcaaatgc atgctcttca acctcaataa cgttatatcc tgattcacgt
     8101 aatatttttg gggtaaattt agttcctgtt ccattaactg cgctaaaaat aatttttaaa
     8161 tcttttttag cttcttgctc ttttttgtac gtctctgttt tagagctaga aatagcaagt
     8221 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttga
     8281 attcgctagc taggtcttga aaggagtggg aattggctcc ggtgcccgtc agtgggcaga
     8341 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gatccggtgc
     8401 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     8461 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     8521 caacgggttt gccgccagaa cacaggaccg gttctagagc gctgccacca tggacaagaa
     8581 gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga
     8641 gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa
     8701 gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct
     8761 gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga
     8821 gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc
     8881 cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga
     8941 cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga
     9001 cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt
     9061 ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct
     9121 gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag
     9181 cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa
     9241 tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct
     9301 gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact
     9361 gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga
     9421 ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga
     9481 catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag
     9541 atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc
     9601 tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga
     9661 cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga
     9721 cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac
     9781 cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg
     9841 gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct
     9901 gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg
     9961 gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa
    10021 gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa
    10081 cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct
    10141 gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca
    10201 gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct
    10261 gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga
    10321 agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa
    10381 ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac
    10441 actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga
    10501 cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg
    10561 gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa
    10621 gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt
    10681 taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat
    10741 tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt
    10801 ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc
    10861 cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat
    10921 cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac
    10981 ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt
    11041 ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca
    11101 gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg
    11161 gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg
    11221 gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga
    11281 gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac
    11341 ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga
    11401 cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc
    11461 cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc
    11521 ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct
    11581 ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa
    11641 gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa
    11701 ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga
    11761 gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg
    11821 gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg
    11881 cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa
    11941 ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct
    12001 ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct
    12061 ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc
    12121 caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt
    12181 cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa
    12241 cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa
    12301 gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca
    12361 ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga
    12421 cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga
    12481 gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt
    12541 caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga
    12601 cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca
    12661 gctgggaggc gacaagcgac ctgccgccac aaagaaggct ggacaggcta agaagaagaa
    12721 agattacaaa gacgatgacg ataagggatc cggcgcaaca aacttctctc tgctgaaaca
    12781 agccggagat gtcgaagaga atcctggacc gattgaacaa gatggattgc acgcaggttc
    12841 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
    12901 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
    12961 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
    13021 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
    13081 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
    13141 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
    13201 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
    13261 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
    13321 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
    13381 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
    13441 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
    13501 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
    13561 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgaacgcgtt aagtcgacaa
    13621 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
    13681 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
    13741 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
    13801 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
    13861 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
    13921 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
    13981 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
    14041 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
    14101 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
    14161 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcgtc gactttaaga
    14221 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
    14281 gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc
    14341 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
    14401 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
    14461 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagggcccgt
    14521 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
    14581 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
    14641 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
    14701 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
    14761 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
    14821 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
    14881 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
    14941 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
    15001 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
    15061 ctgatagacg gt
//
