LOCUS       sequence-358004-       14676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Variant of lentiCRISPRv2 that confers blasticidin S resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14676)
  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 14676)
  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 blast
FEATURES             Location/Qualifiers
     source          1..14676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     25..46
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(165..185)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        170..499
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      350..485
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     412..431
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        547..594
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             613..987
                     /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    1117..1238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1154..1173)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1208..1227
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1287..1303)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1287..1303)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1300..1322)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1311..1327
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1335..1341,1342..1359,1360..1365))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1380..1401
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1518..1535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1689..2277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1769..1788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2448..3239,3240..3308))
                     /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     3071..3090
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3309..3413)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3488..3507)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3683..4062
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4064..4262
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4217..4237
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4280..4460
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4507..4632
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5125..5358
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5543..5587
                     /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    5885..6002
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6053..6301
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6053..6073
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6224..6243
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8183..8258
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8320..8531
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      8550..8559
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8556..12659
                     /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             8556..12659
                     /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             12660..12707
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             12708..12731
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             12741..12797
                     /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             12798..13193
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    13209..13797
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(13262..13282)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(13680..13691)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             13869..14102
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(14128..14145)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    14134..14358
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(14404..14676,1..156)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14491..14510)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
       61 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
      121 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
      181 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
      241 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
      301 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
      361 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
      421 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
      481 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
      541 gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
      601 aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
      661 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
      721 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
      781 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
      841 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
      901 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
      961 cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc
     1021 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     1081 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     1141 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1201 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     1261 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1321 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1381 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1441 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1501 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1561 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1621 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1681 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1741 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     1801 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1861 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1921 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1981 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2041 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2101 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2161 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2221 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2281 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2341 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2401 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2461 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2521 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2581 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2641 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2701 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     2761 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     2821 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     2881 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     2941 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3001 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3061 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3121 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3181 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3241 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3301 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3361 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3421 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     3481 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     3541 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     3601 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     3661 gggccagata tacgcgcgcg ttgacattga ttattgacta gttattaata gtaatcaatt
     3721 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     3781 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     3841 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     3901 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     3961 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     4021 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     4081 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     4141 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     4201 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     4261 agcgcgtttt gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     4321 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     4381 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     4441 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
     4501 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
     4561 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
     4621 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
     4681 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     4741 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     4801 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     4861 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
     4921 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat
     4981 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     5041 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     5101 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     5161 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     5221 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     5281 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     5341 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     5401 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     5461 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     5521 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     5581 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     5641 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     5701 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     5761 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
     5821 attagtgaac ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc
     5881 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     5941 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     6001 ttcgggttta ttacagggac agcagagatc cagtttggtt aattaaggta ccgagggcct
     6061 atttcccatg attccttcat atttgcatat acgatacaag gctgttagag agataattag
     6121 aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata
     6181 atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac
     6241 cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac
     6301 cggagacggt tgtaaatgag cacacaaaat acacatgcta aaatattata ttctatgacc
     6361 tttataaaat caaccaaaat cttcttttta ataactttag tatcaataat tagaattttt
     6421 atgttccttt ttgcaaactt ttaataaaaa tgagcaaaat aaaaaaacgc tagttttagt
     6481 aactcgcgtt gttttcttca cctttaataa tagctactcc accacttgtt cctaagcggt
     6541 cagctcctgc ttcaatcatt ttttgagcat cttcaaatgt tctaactcca ccagctgctt
     6601 taactaaagc attgtcttta acaactgact tcattagttt aacatcttca aatgttgcac
     6661 ctgattttga aaatcctgtt gatgttttaa caaattctaa tccagcttca acagctattt
     6721 cacaagcttt catgatttct tcttttgtta ataaacaatt ttccataata catttaacaa
     6781 catgtgatcc agctgctttt tttacagctt tcatgtcttc taaaactaat tcataatttt
     6841 tgtcttttaa tgcaccaata tttaatacca tatcaatttc tgttgcacca tctttaattg
     6901 cttcagaaac ttcgaatgct tttgtagctg ttgtgcatgc acctagagga aaacctacaa
     6961 catttgttat tcctacattt gtgcctttta ataattcttt acaatagctt gttcaatatg
     7021 aattaacaca aactgttgca aaatcaaatt caattgcttc atcacataat tgtttaattt
     7081 cagctttcgt agcatcttgt tttaataatg tgtgatctat atatttgttt agtttcattt
     7141 tttctcctat atattcattt ttaattttaa ttctttaata atttcgtcta ctttaacttt
     7201 agcgttttga acagattcac caacacctat aaaataaatt tttagtttag gttcagttcc
     7261 acttgggcga acagcaaatc atgacttatc ttctaaataa aattttagta agtcttgtcc
     7321 tggcatatta tacattccat cgatgtagtc ttcaacatta acaactttaa gtccagcaat
     7381 ttgagttaag ggtgttgctc tcaatgattt cattaatggt tcaattttta atttcttttc
     7441 ttctggttta aaattcaagt ttaaagtgaa agtgtaatat gcacccattt ctttaaataa
     7501 atcttctaaa tagtctacta atgttttatt ttgtttttta taaaatcaag cagcctctgc
     7561 tattaatata gaagcttgta ttccatcttt atctctagct gagtcatcaa ttacatatcc
     7621 ataactttct tcataagcaa aaacaaaatt taatccgtta tcttcttctt tagcaatttc
     7681 tctacccatt catttaaatc cagttaaagt ttttacaata ttaactccat atttttcatg
     7741 agcgattcta tcacccaaat cacttgttac aaaacttgaa tatagagccg gattttttgg
     7801 aatgctattt aagcgtttta gatttgataa ttttcaatca attaaaattg gtcctgtttg
     7861 atttccatct aatcttacaa aatgaccatc atgttttatt gccattccaa atctgtcagc
     7921 atctgggtca ttcataataa taatatctgc atcatgttta ataccatatt caagcggtat
     7981 ttttcatgca ggatcaaatt ctggatttgg atttacaaca tttttaaatg tttcatcttc
     8041 aaatgcatgc tcttcaacct caataacgtt atatcctgat tcacgtaata tttttggggt
     8101 aaatttagtt cctgttccat taactgcgct aaaaataatt tttaaatctt ttttagcttc
     8161 ttgctctttt ttgtacgtct ctgttttaga gctagaaata gcaagttaaa ataaggctag
     8221 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttttgaattc gctagctagg
     8281 tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca
     8341 cagtccccga gaagttgggg ggaggggtcg gcaattgatc cggtgcctag agaaggtggc
     8401 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
     8461 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
     8521 ccagaacaca ggaccggttc tagagcgctg ccaccatgga caagaagtac agcatcggcc
     8581 tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
     8641 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
     8701 gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca
     8761 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
     8821 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
     8881 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
     8941 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
     9001 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
     9061 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
     9121 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
     9181 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc
     9241 tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga
     9301 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg
     9361 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
     9421 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
     9481 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc
     9541 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
     9601 agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc
     9661 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac
     9721 tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca
     9781 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt
     9841 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc
     9901 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga
     9961 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc
    10021 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc
    10081 ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat
    10141 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg
    10201 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
    10261 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
    10321 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
    10381 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
    10441 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
    10501 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
    10561 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
    10621 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
    10681 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
    10741 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
    10801 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
    10861 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
    10921 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
    10981 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
    11041 acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg
    11101 acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca
    11161 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg
    11221 ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga
    11281 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
    11341 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc
    11401 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
    11461 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
    11521 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg
    11581 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
    11641 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
    11701 agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa
    11761 ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca
    11821 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt
    11881 ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta
    11941 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag
    12001 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca
    12061 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag
    12121 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg
    12181 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc
    12241 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc
    12301 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga
    12361 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca
    12421 aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata
    12481 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca
    12541 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc
    12601 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca
    12661 agcgacctgc cgccacaaag aaggctggac aggctaagaa gaagaaagat tacaaagacg
    12721 atgacgataa gggatccggc gcaacaaact tctctctgct gaaacaagcc ggagatgtcg
    12781 aagagaatcc tggaccggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag
    12841 caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc
    12901 tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt
    12961 gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta
    13021 tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg
    13081 tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga
    13141 cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taaacgcgtt
    13201 aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
    13261 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
    13321 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
    13381 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
    13441 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
    13501 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
    13561 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
    13621 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
    13681 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
    13741 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcgtc
    13801 gactttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
    13861 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
    13921 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
    13981 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
    14041 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
    14101 cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
    14161 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
    14221 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
    14281 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
    14341 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
    14401 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
    14461 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
    14521 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
    14581 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
    14641 ggccatcgcc ctgatagacg gtttttcgcc ctttga
//
