LentiCRISPRv2-ATG5

LOCUS       LentiCRISPRv2-ATG5    13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 with sgRNA targeting Exon 7 of ATG5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  AUTHORS   Imam S, Talley S, Nelson RS, Dharan A, O'Connor C, Hope TJ, Campbell
            EM
  TITLE     TRIM5alpha degradation via autophagy is not required for retroviral 
            restriction.
  JOURNAL   J Virol. 2016 Jan 13. pii: JVI.03033-15.
  PUBMED    26764007
REFERENCE   2  (bases 1 to 13013)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Virol. 
            2016 Jan 13. pii: JVI.03033-15."
COMMENT     Sequence Label: LentiCRISPRv2-ATG5
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..55)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        40..369
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      220..355
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     282..301
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        417..464
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             483..857
                     /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    987..1108
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1024..1043)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1078..1097
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1157..1173)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1157..1173)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1170..1192)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1181..1197
                     /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(1205..1211,1212..1229,1230..1235))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1250..1271
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1388..1405)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1559..2147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1639..1658)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2318..3109,3110..3178))
                     /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     2941..2960
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3179..3283)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3358..3377)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3549..3928
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3930..4128
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4083..4103
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4146..4326
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4373..4498
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4991..5224
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5409..5453
                     /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    5751..5868
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5919..6167
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5919..5939
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6090..6109
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6189..6264
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        6326..6537
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6556..6565
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6562..10665
                     /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             6562..10665
                     /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             10666..10713
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             10714..10737
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             10747..10803
                     /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             10804..11400
                     /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     11297..11317
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    11416..12004
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11469..11489)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11887..11898)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             12076..12309
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(12335..12352)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12341..12565
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(12611..13013,1..26)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12698..12717)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12908..12929
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
       61 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
      121 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
      181 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
      241 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
      301 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
      361 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg
      421 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
      481 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
      541 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
      601 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
      661 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
      721 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
      781 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
      841 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
      901 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
      961 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     1021 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1081 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     1141 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     1201 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     1261 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     1321 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1381 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1441 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1501 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1561 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1621 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1681 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1741 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1801 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1861 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1921 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1981 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2041 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2101 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2161 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2221 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2281 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2341 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2401 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2461 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2521 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2581 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2641 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2701 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2761 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2821 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     2881 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     2941 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3001 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3061 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3121 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3181 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3241 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3301 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     3361 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     3421 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     3481 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     3541 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     3601 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     3661 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     3721 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     3781 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     3841 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     3901 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     3961 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     4021 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     4081 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg cgttttgcct
     4141 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     4201 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     4261 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     4321 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
     4381 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
     4441 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
     4501 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
     4561 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
     4621 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
     4681 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
     4741 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
     4801 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag
     4861 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
     4921 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
     4981 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
     5041 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
     5101 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     5161 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     5221 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     5281 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     5341 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     5401 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     5461 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     5521 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     5581 attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag
     5641 gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat
     5701 cggcactgcg tgcgccaatt ctgcagacaa atggcagtat tcatccacaa ttttaaaaga
     5761 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     5821 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac
     5881 agggacagca gagatccagt ttggttaatt aaggtaccga gggcctattt cccatgattc
     5941 cttcatattt gcatatacga tacaaggctg ttagagagat aattagaatt aatttgactg
     6001 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt
     6061 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt
     6121 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccgga tggacagttg
     6181 cacacactgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
     6241 gaaaaagtgg caccgagtcg gtgctttttt gaattcgcta gctaggtctt gaaaggagtg
     6301 ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     6361 ttggggggag gggtcggcaa ttgatccggt gcctagagaa ggtggcgcgg ggtaaactgg
     6421 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     6481 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggac
     6541 cggttctaga gcgctgccac catggacaag aagtacagca tcggcctgga catcggcacc
     6601 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
     6661 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
     6721 gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
     6781 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
     6841 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
     6901 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
     6961 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
     7021 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
     7081 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
     7141 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
     7201 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
     7261 aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
     7321 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
     7381 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
     7441 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
     7501 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
     7561 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
     7621 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
     7681 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
     7741 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
     7801 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
     7861 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
     7921 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
     7981 accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag
     8041 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
     8101 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
     8161 atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc
     8221 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
     8281 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
     8341 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
     8401 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
     8461 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
     8521 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
     8581 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
     8641 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
     8701 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
     8761 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
     8821 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
     8881 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
     8941 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
     9001 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
     9061 tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac
     9121 aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
     9181 gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc
     9241 cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
     9301 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
     9361 atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg
     9421 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
     9481 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
     9541 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
     9601 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
     9661 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
     9721 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg
     9781 tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat
     9841 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag
     9901 aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
     9961 ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
    10021 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
    10081 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
    10141 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
    10201 ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg
    10261 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
    10321 cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc
    10381 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc
    10441 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
    10501 accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg
    10561 aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
    10621 ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaagcg acctgccgcc
    10681 acaaagaagg ctggacaggc taagaagaag aaagattaca aagacgatga cgataaggga
    10741 tccggcgcaa caaacttctc tctgctgaaa caagccggag atgtcgaaga gaatcctgga
    10801 ccgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
    10861 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
    10921 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
    10981 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
    11041 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
    11101 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
    11161 cccgcgtggt tcctggccac cgtcggagtc tcgcccgacc accagggcaa gggtctgggc
    11221 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
    11281 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
    11341 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
    11401 acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
    11461 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
    11521 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
    11581 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
    11641 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
    11701 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
    11761 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
    11821 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
    11881 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
    11941 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
    12001 ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
    12061 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg
    12121 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
    12181 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
    12241 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
    12301 tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
    12361 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
    12421 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
    12481 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
    12541 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
    12601 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
    12661 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
    12721 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
    12781 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
    12841 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
    12901 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
    12961 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gct
//