LOCUS       Exported               14877 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Variant of lentiCRISPRv2 that confers puromycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    lentiCRISPRv2 puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14877)
  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 14877)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14877
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        242..621
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        623..821
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     776..796
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             839..1019
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1066..1191
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1684..1917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2102..2146
                     /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    2444..2561
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2612..2852
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     2612..2632
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2783..2802
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4742..4817
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4879..5090
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5109..5118
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             5115..9218
                     /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             9219..9266
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9267..9290
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9300..9356
                     /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             9357..9953
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     9850..9870
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9969..10557
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10022..10042)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10440..10451)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10629..10862
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10888..10905)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10894..11118
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11164..11592
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11251..11270)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11461..11482
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11601..11621)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11606..11935
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11786..11921
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11848..11867
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11983..12030
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12049..12423
                     /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    12553..12674
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12590..12609)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12644..12663
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12723..12739)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12723..12739)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12736..12758)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12747..12763
                     /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(12771..12801)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12816..12837
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12954..12971)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13125..13713)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13205..13224)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(13884..14744)
                     /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     14507..14526
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14745..14849)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgcgcgt
      241 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
      301 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      361 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      421 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      481 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      541 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      601 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      661 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      721 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      781 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg
      841 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
      901 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
      961 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     1021 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
     1081 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
     1141 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
     1201 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
     1261 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
     1321 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
     1381 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
     1441 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
     1501 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga
     1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     2581 gcagagatcc agtttggtta attaaggtac cgagggccta tttcccatga ttccttcata
     2641 tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac
     2701 aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt
     2761 tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga
     2821 tttcttggct ttatatatct tgtggaaagg acgaaacacc ggagacggtt gtaaatgagc
     2881 acacaaaata cacatgctaa aatattatat tctatgacct ttataaaatc aaccaaaatc
     2941 ttctttttaa taactttagt atcaataatt agaattttta tgttcctttt tgcaaacttt
     3001 taataaaaat gagcaaaata aaaaaacgct agttttagta actcgcgttg ttttcttcac
     3061 ctttaataat agctactcca ccacttgttc ctaagcggtc agctcctgct tcaatcattt
     3121 tttgagcatc ttcaaatgtt ctaactccac cagctgcttt aactaaagca ttgtctttaa
     3181 caactgactt cattagttta acatcttcaa atgttgcacc tgattttgaa aatcctgttg
     3241 atgttttaac aaattctaat ccagcttcaa cagctatttc acaagctttc atgatttctt
     3301 cttttgttaa taaacaattt tccataatac atttaacaac atgtgatcca gctgcttttt
     3361 ttacagcttt catgtcttct aaaactaatt cataattttt gtcttttaat gcaccaatat
     3421 ttaataccat atcaatttct gttgcaccat ctttaattgc ttcagaaact tcgaatgctt
     3481 ttgtagctgt tgtgcatgca cctagaggaa aacctacaac atttgttatt cctacatttg
     3541 tgccttttaa taattcttta caatagcttg ttcaatatga attaacacaa actgttgcaa
     3601 aatcaaattc aattgcttca tcacataatt gtttaatttc agctttcgta gcatcttgtt
     3661 ttaataatgt gtgatctata tatttgttta gtttcatttt ttctcctata tattcatttt
     3721 taattttaat tctttaataa tttcgtctac tttaacttta gcgttttgaa cagattcacc
     3781 aacacctata aaataaattt ttagtttagg ttcagttcca cttgggcgaa cagcaaatca
     3841 tgacttatct tctaaataaa attttagtaa gtcttgtcct ggcatattat acattccatc
     3901 gatgtagtct tcaacattaa caactttaag tccagcaatt tgagttaagg gtgttgctct
     3961 caatgatttc attaatggtt caatttttaa tttcttttct tctggtttaa aattcaagtt
     4021 taaagtgaaa gtgtaatatg cacccatttc tttaaataaa tcttctaaat agtctactaa
     4081 tgttttattt tgttttttat aaaatcaagc agcctctgct attaatatag aagcttgtat
     4141 tccatcttta tctctagctg agtcatcaat tacatatcca taactttctt cataagcaaa
     4201 aacaaaattt aatccgttat cttcttcttt agcaatttct ctacccattc atttaaatcc
     4261 agttaaagtt tttacaatat taactccata tttttcatga gcgattctat cacccaaatc
     4321 acttgttaca aaacttgaat atagagccgg attttttgga atgctattta agcgttttag
     4381 atttgataat tttcaatcaa ttaaaattgg tcctgtttga tttccatcta atcttacaaa
     4441 atgaccatca tgttttattg ccattccaaa tctgtcagca tctgggtcat tcataataat
     4501 aatatctgca tcatgtttaa taccatattc aagcggtatt tttcatgcag gatcaaattc
     4561 tggatttgga tttacaacat ttttaaatgt ttcatcttca aatgcatgct cttcaacctc
     4621 aataacgtta tatcctgatt cacgtaatat ttttggggta aatttagttc ctgttccatt
     4681 aactgcgcta aaaataattt ttaaatcttt tttagcttct tgctcttttt tgtacgtctc
     4741 tgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
     4801 tggcaccgag tcggtgcttt tttgaattcg ctagctaggt cttgaaagga gtgggaattg
     4861 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     4921 gaggggtcgg caattgatcc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     4981 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     5041 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gaccggttct
     5101 agagcgctgc caccatggac aagaagtaca gcatcggcct ggacatcggc accaactctg
     5161 tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg
     5221 gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg
     5281 gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga
     5341 agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca
     5401 gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc
     5461 accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct
     5521 accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc
     5581 tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc
     5641 ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt
     5701 tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac
     5761 tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg
     5821 gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact
     5881 tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg
     5941 acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc
     6001 tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc
     6061 ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga
     6121 aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca
     6181 agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca
     6241 tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag
     6301 aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc
     6361 tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca
     6421 accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg
     6481 ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct
     6541 ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga
     6601 ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg
     6661 agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa
     6721 agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca
     6781 accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg
     6841 actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg
     6901 atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc
     6961 tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc
     7021 tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat
     7081 acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg
     7141 gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc
     7201 tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc
     7261 agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg
     7321 gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc
     7381 ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga
     7441 acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc
     7501 tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc
     7561 tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact
     7621 acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg
     7681 tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg
     7741 tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa
     7801 agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct
     7861 tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg
     7921 actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga
     7981 tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc
     8041 gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg
     8101 ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt
     8161 acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt
     8221 acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg
     8281 agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata
     8341 agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga
     8401 aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca
     8461 gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca
     8521 gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga
     8581 aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga
     8641 agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca
     8701 tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct
     8761 ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc
     8821 tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac
     8881 agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt
     8941 tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca
     9001 agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga
     9061 ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt
     9121 acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt
     9181 acgagacacg gatcgacctg tctcagctgg gaggcgacaa gcgacctgcc gccacaaaga
     9241 aggctggaca ggctaagaag aagaaagatt acaaagacga tgacgataag ggatccggcg
     9301 caacaaactt ctctctgctg aaacaagccg gagatgtcga agagaatcct ggaccgaccg
     9361 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     9421 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     9481 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     9541 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     9601 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     9661 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     9721 ggttcctggc caccgtcgga gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     9781 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     9841 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     9901 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgaacgcgtt
     9961 aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
    10021 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
    10081 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
    10141 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
    10201 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
    10261 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
    10321 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
    10381 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
    10441 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
    10501 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcgtc
    10561 gactttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
    10621 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
    10681 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
    10741 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
    10801 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
    10861 cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
    10921 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
    10981 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
    11041 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
    11101 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
    11161 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
    11221 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
    11281 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
    11341 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
    11401 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
    11461 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
    11521 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
    11581 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
    11641 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
    11701 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
    11761 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
    11821 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
    11881 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
    11941 cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg
    12001 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg
    12061 accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc
    12121 gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac
    12181 gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc
    12241 tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg
    12301 aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg
    12361 gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac
    12421 tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
    12481 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
    12541 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
    12601 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
    12661 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
    12721 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
    12781 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
    12841 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
    12901 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
    12961 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
    13021 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
    13081 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
    13141 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
    13201 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
    13261 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
    13321 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
    13381 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
    13441 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
    13501 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
    13561 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
    13621 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
    13681 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
    13741 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
    13801 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
    13861 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
    13921 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
    13981 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
    14041 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
    14101 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
    14161 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
    14221 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
    14281 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
    14341 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
    14401 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
    14461 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
    14521 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
    14581 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
    14641 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
    14701 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
    14761 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
    14821 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgac
//
