LOCUS       sequence-406405-       14757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pLentiCRISPRv2 variant with mU6 promoter driving the sgRNA, and a 
            Blasticidin resistance cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14757)
  AUTHORS   Unlu B, Pons C, Ho UL, Batte A, Aloy P, van Leeuwen J
  TITLE     Global analysis of suppressor mutations that rescue human genetic 
            defects.
  JOURNAL   Genome Med. 2023 Oct 12;15(1):78. doi: 10.1186/s13073-023-01232-0.
  PUBMED    37821946
REFERENCE   2  (bases 1 to 14757)
  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: "Genome 
            Med."; date: "2023-10-12"; volume: "15(1):78. doi"; pages: " 
            10.1186/s13073-023-01232-0"
COMMENT     Sequence Label: lentiCRISPRv2-Blast-mU6
FEATURES             Location/Qualifiers
     source          1..14757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(84..104)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        89..418
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      269..404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     331..350
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        466..513
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             532..906
                     /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    1036..1157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1073..1092)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1127..1146
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1206..1222)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1206..1222)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1219..1241)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1230..1246
                     /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(1254..1260,1261..1278,1279..1284))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1299..1320
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1437..1454)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1608..2196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1688..1707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2367..3158,3159..3227))
                     /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     2990..3009
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3228..3332)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3407..3426)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3598..3977
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3979..4177
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4132..4152
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4195..4375
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4422..4547
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5040..5273
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5458..5502
                     /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    5800..5917
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5968..6281
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     6200..6220
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     misc_RNA        8171..8246
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8308..8519
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      8538..8547
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8544..12647
                     /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             8544..12647
                     /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             12648..12695
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             12696..12719
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             12729..12785
                     /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             12786..13184
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     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..14757,1..75)
                     /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"
     primer_bind     14701..14722
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
       61 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
      121 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
      181 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      241 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
      301 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
      361 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
      421 gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca
      481 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag
      541 ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg
      601 accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg
      661 gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg
      721 gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc
      781 acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg
      841 cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag
      901 gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
      961 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     1021 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1081 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1141 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     1201 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1261 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1321 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     1381 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     1441 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     1501 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     1561 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1621 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1681 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1741 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1801 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1861 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1921 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1981 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2041 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2101 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2161 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2221 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2281 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2341 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2401 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     2461 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     2521 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     2581 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     2641 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     2701 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     2761 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2821 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2881 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2941 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3001 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3061 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3121 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3181 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3241 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3301 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     3361 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     3421 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     3481 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     3541 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
     3601 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     3661 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     3721 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     3781 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     3841 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     3901 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     3961 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     4021 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     4081 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     4141 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
     4201 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     4261 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     4321 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     4381 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     4441 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     4501 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     4561 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     4621 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     4681 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     4741 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     4801 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     4861 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     4921 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     4981 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     5041 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     5101 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     5161 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     5221 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     5281 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     5341 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     5401 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     5461 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     5521 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     5581 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     5641 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     5701 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     5761 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     5821 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     5881 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     5941 agatccagtt tggttaatta aggtaccgat ccgacgccgc catctctagg cccgcgccgg
     6001 ccccctcgca cagacttgtg ggagaagctc ggctactccc ctgccccggt taatttgcat
     6061 ataatatttc ctagtaacta tagaggctta atgtgcgata aaagacagat aatctgttct
     6121 ttttaatact agctacattt tacatgatag gcttggattt ctataagaga tacaaatact
     6181 aaattattat tttaaaaaac agcacaaaag gaaactcacc ctaactgtaa agtaattgtg
     6241 tgttttgaga ctataaatat cccttggaga aaagccttgt tgaaacaccg gagacggttg
     6301 taaatgagca cacaaaatac acatgctaaa atattatatt ctatgacctt tataaaatca
     6361 accaaaatct tctttttaat aactttagta tcaataatta gaatttttat gttccttttt
     6421 gcaaactttt aataaaaatg agcaaaataa aaaaacgcta gttttagtaa ctcgcgttgt
     6481 tttcttcacc tttaataata gctactccac cacttgttcc taagcggtca gctcctgctt
     6541 caatcatttt ttgagcatct tcaaatgttc taactccacc agctgcttta actaaagcat
     6601 tgtctttaac aactgacttc attagtttaa catcttcaaa tgttgcacct gattttgaaa
     6661 atcctgttga tgttttaaca aattctaatc cagcttcaac agctatttca caagctttca
     6721 tgatttcttc ttttgttaat aaacaatttt ccataataca tttaacaaca tgtgatccag
     6781 ctgctttttt tacagctttc atgtcttcta aaactaattc ataatttttg tcttttaatg
     6841 caccaatatt taataccata tcaatttctg ttgcaccatc tttaattgct tcagaaactt
     6901 cgaatgcttt tgtagctgtt gtgcatgcac ctagaggaaa acctacaaca tttgttattc
     6961 ctacatttgt gccttttaat aattctttac aatagcttgt tcaatatgaa ttaacacaaa
     7021 ctgttgcaaa atcaaattca attgcttcat cacataattg tttaatttca gctttcgtag
     7081 catcttgttt taataatgtg tgatctatat atttgtttag tttcattttt tctcctatat
     7141 attcattttt aattttaatt ctttaataat ttcgtctact ttaactttag cgttttgaac
     7201 agattcacca acacctataa aataaatttt tagtttaggt tcagttccac ttgggcgaac
     7261 agcaaatcat gacttatctt ctaaataaaa ttttagtaag tcttgtcctg gcatattata
     7321 cattccatcg atgtagtctt caacattaac aactttaagt ccagcaattt gagttaaggg
     7381 tgttgctctc aatgatttca ttaatggttc aatttttaat ttcttttctt ctggtttaaa
     7441 attcaagttt aaagtgaaag tgtaatatgc acccatttct ttaaataaat cttctaaata
     7501 gtctactaat gttttatttt gttttttata aaatcaagca gcctctgcta ttaatataga
     7561 agcttgtatt ccatctttat ctctagctga gtcatcaatt acatatccat aactttcttc
     7621 ataagcaaaa acaaaattta atccgttatc ttcttcttta gcaatttctc tacccattca
     7681 tttaaatcca gttaaagttt ttacaatatt aactccatat ttttcatgag cgattctatc
     7741 acccaaatca cttgttacaa aacttgaata tagagccgga ttttttggaa tgctatttaa
     7801 gcgttttaga tttgataatt ttcaatcaat taaaattggt cctgtttgat ttccatctaa
     7861 tcttacaaaa tgaccatcat gttttattgc cattccaaat ctgtcagcat ctgggtcatt
     7921 cataataata atatctgcat catgtttaat accatattca agcggtattt ttcatgcagg
     7981 atcaaattct ggatttggat ttacaacatt tttaaatgtt tcatcttcaa atgcatgctc
     8041 ttcaacctca ataacgttat atcctgattc acgtaatatt tttggggtaa atttagttcc
     8101 tgttccatta actgcgctaa aaataatttt taaatctttt ttagcttctt gctctttttt
     8161 gtacgtctct gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac
     8221 ttgaaaaagt ggcaccgagt cggtgctttt ttgaattcgc tagctaggtc ttgaaaggag
     8281 tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
     8341 agttgggggg aggggtcggc aattgatccg gtgcctagag aaggtggcgc ggggtaaact
     8401 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
     8461 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg
     8521 accggttcta gagcgctgcc accatggaca agaagtacag catcggcctg gacatcggca
     8581 ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca
     8641 aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga gccctgctgt
     8701 tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca
     8761 ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg
     8821 tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc
     8881 acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc
     8941 ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc
     9001 tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg
     9061 acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca
     9121 accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt
     9181 ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga
     9241 agaagaatgg cctgttcgga aacctgattg ccctgagcct gggcctgacc cccaacttca
     9301 agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg
     9361 acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg
     9421 ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca
     9481 ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga
     9541 ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg
     9601 accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct
     9661 acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc
     9721 tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc
     9781 agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc
     9841 tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg
     9901 gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca
     9961 tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag agcttcatcg
    10021 agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc
    10081 tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg
    10141 gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt
    10201 tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg
    10261 agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca
    10321 cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg
    10381 aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg
    10441 aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc
    10501 ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca
    10561 agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact
    10621 tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg
    10681 tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca
    10741 ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc
    10801 ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg
    10861 gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca
    10921 gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc
    10981 tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc
    11041 tgtccgacta cgatgtggac catatcgtgc ctcagagctt tctgaaggac gactccatcg
    11101 acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg
    11161 aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta
    11221 cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata
    11281 aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac
    11341 agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag
    11401 tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt
    11461 acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg
    11521 tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact
    11581 acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta
    11641 ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg
    11701 ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg
    11761 tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga
    11821 atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca
    11881 agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg
    11941 gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca
    12001 agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca
    12061 gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg
    12121 acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa
    12181 tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg
    12241 tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg
    12301 agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga
    12361 tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg
    12421 cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt
    12481 ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc
    12541 ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca
    12601 ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacaag cgacctgccg
    12661 ccacaaagaa ggctggacag gctaagaaga agaaagatta caaagacgat gacgataagg
    12721 gatccggcgc aacaaacttc tctctgctga aacaagccgg agatgtcgaa gagaatcctg
    12781 gaccgatggc caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta
    12841 caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg
    12901 acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac
    12961 tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga
    13021 tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg
    13081 atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg
    13141 ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaagaattc gatatcaagc
    13201 ttatcggtaa 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 ctttgacgtt ggagtccacg ttctttaata
    14701 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttg
//
