LOCUS       sequence-322376-       12859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized S. pyogenes Cas9 protein and 
            blasticidin resistance from EFS promoter. 3rd generation lentiviral 
            backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12859)
  AUTHORS   Sanjana NE, Shalem O, Zhang F
  TITLE     Improved vectors and genome-wide libraries for CRISPR screening.
  JOURNAL   Nat Methods. 2014 Aug;11(8):783-4. doi: 10.1038/nmeth.3047.
  PUBMED    25075903
REFERENCE   2  (bases 1 to 12859)
  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"; doi: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2014-08"; volume: "11"; issue: 
            "8"; pages: "783-4"
COMMENT     Sequence Label: lentiCas9-Blast
FEATURES             Location/Qualifiers
     source          1..12859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             119..299
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(325..342)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    331..555
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      601..1029
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(688..707)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     898..919
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1038..1058)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1043..1372
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1223..1358
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1285..1304
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1420..1467
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1486..1860
                     /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    1990..2111
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2027..2046)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2081..2100
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2160..2176)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2160..2176)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2173..2195)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2184..2200
                     /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(2208..2214,2215..2232,2233..2238))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2253..2274
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2391..2408)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2562..3150)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2642..2661)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3321..4112,4113..4181))
                     /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     3944..3963
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4182..4286)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4361..4380)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4552..4931
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4933..5131
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5086..5106
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5149..5329
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5376..5501
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5994..6227
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6412..6456
                     /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    6754..6871
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6964..7175
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      7194..7203
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7200..11303
                     /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             7200..11303
                     /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             11304..11351
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             11352..11375
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             11385..11441
                     /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             11442..11840
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    11865..12453
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11918..11938)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(12336..12347)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(12456..12472)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12457..12473)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat tagagtggag
       61 gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg actgtactgg
      121 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
      181 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
      241 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
      301 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
      361 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
      421 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
      481 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
      541 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
      601 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
      661 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      721 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      781 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      841 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      901 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      961 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     1021 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     1081 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     1141 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     1201 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     1261 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     1321 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     1381 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
     1441 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc
     1501 agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac
     1561 cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac
     1621 gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg
     1681 gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac
     1741 ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag
     1801 ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga
     1861 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     1921 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     1981 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2041 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2101 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     2161 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     2221 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2281 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2341 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2401 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2461 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2521 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2581 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2641 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2701 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2761 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2821 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2881 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2941 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3001 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3061 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3121 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3181 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3241 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3301 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3361 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3421 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3481 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3541 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3601 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3661 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3721 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3781 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3841 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3901 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3961 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4021 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4081 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4141 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4201 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4261 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     4321 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     4381 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     4441 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     4501 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
     4561 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     4621 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     4681 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     4741 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     4801 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     4861 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     4921 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     4981 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     5041 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     5101 ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg
     5161 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     5221 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     5281 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
     5341 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
     5401 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
     5461 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
     5521 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
     5581 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
     5641 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
     5701 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     5761 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     5821 agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac
     5881 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     5941 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     6001 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     6061 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     6121 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     6181 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     6241 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     6301 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     6361 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     6421 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     6481 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     6541 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     6601 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
     6661 ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact gcgtgcgcca
     6721 attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg
     6781 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
     6841 acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc
     6901 agtttggtta attagctagc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
     6961 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
     7021 gatccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
     7081 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
     7141 ttctttttcg caacgggttt gccgccagaa cacaggaccg gttctagagc gctgccacca
     7201 tggacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga
     7261 tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc
     7321 acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg
     7381 ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct
     7441 atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac
     7501 tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca
     7561 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga
     7621 aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca
     7681 tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg
     7741 tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca
     7801 tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac
     7861 ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc
     7921 tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg
     7981 atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc
     8041 agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc
     8101 tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta
     8161 tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc
     8221 agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg
     8281 gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg
     8341 aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga
     8401 agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg
     8461 ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg
     8521 agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca
     8581 gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag
     8641 tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga
     8701 acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt
     8761 ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga
     8821 gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg
     8881 tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct
     8941 ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta
     9001 tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc
     9061 tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc
     9121 acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca
     9181 ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg
     9241 atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca
     9301 gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc
     9361 acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag
     9421 tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga
     9481 tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa
     9541 tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg
     9601 tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg
     9661 atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata
     9721 tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg
     9781 acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga
     9841 actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga
     9901 ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc
     9961 tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca
    10021 ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca
    10081 agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact
    10141 accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt
    10201 accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga
    10261 tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca
    10321 acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc
    10381 ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg
    10441 ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc
    10501 agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg
    10561 ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct
    10621 attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga
    10681 aagagctact ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact
    10741 ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt
    10801 actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc
    10861 agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc
    10921 actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac
    10981 agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga
    11041 tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc
    11101 ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc
    11161 ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag
    11221 aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg
    11281 acctgtctca gctgggaggc gacaagcgac ctgccgccac aaagaaggct ggacaggcta
    11341 agaagaagaa agattacaaa gacgatgacg ataagggatc cggcgcaaca aacttctctc
    11401 tgctgaaaca agccggagat gtcgaagaga atcctggacc gatggccaag cctttgtctc
    11461 aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg
    11521 aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca
    11581 atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg
    11641 ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct
    11701 tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca
    11761 tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg
    11821 gttatgtgtg ggagggctaa gaattcgata tcaagcttat cggtaatcaa cctctggatt
    11881 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
    11941 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
    12001 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
    12061 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
    12121 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
    12181 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
    12241 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
    12301 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
    12361 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
    12421 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct cgagacctag
    12481 aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc
    12541 tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac
    12601 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
    12661 aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac
    12721 aaggctactt ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga
    12781 cctttggatg gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg
    12841 aaggagagaa cacccgctt
//
