LOCUS       sequence-129399-       12705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing Cas9 fused with the Blasticidin 
            resistant gene at the N-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12705)
  AUTHORS   Tzelepis K, Koike-Yusa H, De Braekeleer E, Li Y, Metzakopian E, 
            Dovey OM, Mupo A, Grinkevich V, Li M, Mazan M, Gozdecka M, Ohnishi 
            S, Cooper J, Patel M, McKerrell T, Chen B, Domingues AF, Gallipoli 
            P, Teichmann S, Ponstingl H, McDermott U, Saez-Rodriguez J, Huntly 
            BJ, Iorio F, Pina C, Vassiliou GS, Yusa K
  TITLE     A CRISPR Dropout Screen Identifies Genetic Vulnerabilities and 
            Therapeutic Targets in Acute Myeloid Leukemia.
  JOURNAL   Cell Rep. 2016 Oct 18;17(4):1193-1205. doi: 
            10.1016/j.celrep.2016.09.079.
  PUBMED    27760321
REFERENCE   2  (bases 1 to 12705)
  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: 
            "10.1016/j.celrep.2016.09"; journalName: "Cell Rep"; date: 
            "2016-10-18- 18"; volume: "17"; issue: "4"; pages: "1193-1205"
COMMENT     Sequence Label: pKLV2-EF1a-BsdCas9-W
FEATURES             Location/Qualifiers
     source          1..12705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        960..1158
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1113..1133
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1176..1356
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1403..1528
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2027..2260
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2445..2489
                     /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    2787..2904
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3375..4560
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3606..4552
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4509..4529
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4581..4976
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             4995..5048
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5052..5117
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5124..5144
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5169..9269
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5169..9269
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             9270..9317
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(9380..9400)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9798..9809)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10261..10278)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10267..10491
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(10517..10537)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(10517..10535)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10556..10572)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10556..10572)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10569..10591)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10580..10596
                     /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(10604..10610,10611..10628,10629..10634))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10649..10670
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10787..10804)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10958..11546)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11038..11057)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11717..12508,12509..12577))
                     /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     12340..12359
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12578..12682)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgactagtac
      661 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      721 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      781 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      841 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      901 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      961 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     1021 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     1081 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     1141 ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct
     1201 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     1261 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     1321 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     1381 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     1441 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     1501 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     1561 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     1621 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     1681 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     1741 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     1801 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     1861 gcaagcggcc ggccgcgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     1921 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     1981 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     2041 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     2101 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     2161 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     2221 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     2281 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     2341 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     2401 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     2461 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     2521 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     2581 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     2641 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
     2701 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc
     2761 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     2821 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     2881 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg
     2941 ttagtaccgg gccctacgcg tgaggggcgc ggaagccggg gcggaggagg agccaacccc
     3001 gggctgggct gagacccgca gaggaagacg ctctagggat ttgtcccgga ctagcgagat
     3061 ggcaaggctg aggacgggag gctgattgag aggcgaaggt acaccctaat ctcaatacaa
     3121 cctttggagc taagccagca atggtagagg gaagattctg cacgtccctt ccaggcggcc
     3181 tccccgtcac cacccccccc aacccgcccc gaccggagct gagagtaatt catacaaaag
     3241 gactcgcccc tgccttgggg aatcccaggg accgtcgtta aactcccact aacgtagaac
     3301 ccagagatcg ctgcgttccc gccccctcac ccgcccgctc tcgtcatcac tgaggtggag
     3361 aagagcatgc gtgaggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc
     3421 ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg
     3481 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac
     3541 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa
     3601 cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt
     3661 gcgtgccttg aattacttcc acgcccctgg ctgcagtacg tgattcttga tcccgagctt
     3721 cgggttggaa gtgggtggga gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt
     3781 gcttgagttg aggcctggct tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt
     3841 cgcgcctgtc tcgctgcttt cgataagtct ctagccattt aaaatttttg atgacctgct
     3901 gcgacgcttt ttttctggca agatagtctt gtaaatgcgg gccaagatcg atctgcacac
     3961 tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg
     4021 ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc
     4081 tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc
     4141 aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc
     4201 tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac
     4261 acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg
     4321 ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg
     4381 gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag
     4441 gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg
     4501 gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg
     4561 ggaagcttgg cgcgcccacc atggccaagc ctttgtctca agaagaatcc accctcattg
     4621 aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg
     4681 cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg
     4741 gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga
     4801 cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc
     4861 gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac
     4921 agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggcggcg
     4981 gcgggtccgg aggagagggc agaggaagtc tcctaacatg cggtgacgtg gaggagaatc
     5041 ctggcccaat ggactataag gaccacgacg gagactacaa ggatcatgat attgattaca
     5101 aagacgatga cgataagatg gccccaaaga agaagcggaa ggtcggtatc cacggagtcc
     5161 cagcagccga caagaagtac agcatcggcc tggacatcgg caccaactct gtgggctggg
     5221 ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg
     5281 accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag
     5341 ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga
     5401 tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc
     5461 acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct
     5521 tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga
     5581 gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg
     5641 cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca
     5701 gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa
     5761 accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga
     5821 gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg
     5881 gaaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg
     5941 ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc
     6001 tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg
     6061 ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg
     6121 cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg
     6181 tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct
     6241 acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca
     6301 tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc
     6361 tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc
     6421 tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa
     6481 agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa
     6541 acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg
     6601 aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg
     6661 ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca
     6721 ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct
     6781 tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag
     6841 tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg
     6901 aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga
     6961 aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata
     7021 tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct
     7081 atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct
     7141 ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa
     7201 tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg
     7261 acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata
     7321 gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc
     7381 agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca
     7441 tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg
     7501 agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac
     7561 accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg
     7621 ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg
     7681 accatatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca
     7741 gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga
     7801 tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca
     7861 atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga
     7921 gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga
     7981 tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga
     8041 agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca
     8101 acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca
     8161 aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc
     8221 ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct
     8281 acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga
     8341 agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg
     8401 attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg
     8461 aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc
     8521 tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg
     8581 tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga
     8641 gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca
     8701 tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc
     8761 ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg
     8821 aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg
     8881 ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg
     8941 tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga
     9001 gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg
     9061 ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg
     9121 gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca
     9181 ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac
     9241 ggatcgacct gtctcagctg ggaggcgaca aaaggccggc ggccacgaaa aaggccggcc
     9301 aggcaaaaaa gaaaaagtaa gcggccgctc aacctctgga ttacaaaatt tgtgaaagat
     9361 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     9421 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     9481 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     9541 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     9601 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     9661 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     9721 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     9781 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     9841 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     9901 gggccgcctc cccgcaggta cctttaagac caatgactta caaggcagct gtagatctta
     9961 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa agaagtcaag
    10021 atctgctttt tgcctgtact gggtctctct ggttagacca gagtctctct ggttagacca
    10081 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
    10141 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
    10201 atccctcaga cccttttagt cagtgtggaa aatctctagc agtttaaacc cgctgatcag
    10261 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
    10321 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
    10381 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
    10441 aggattggga agtcaatagc aggcatgctg gggatgcggt gggctctatg ggcggccgtt
    10501 aattaagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
    10561 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
    10621 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
    10681 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
    10741 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
    10801 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
    10861 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
    10921 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
    10981 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
    11041 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
    11101 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
    11161 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
    11221 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
    11281 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
    11341 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
    11401 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
    11461 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
    11521 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
    11581 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
    11641 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
    11701 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
    11761 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
    11821 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
    11881 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
    11941 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
    12001 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
    12061 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
    12121 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
    12181 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
    12241 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
    12301 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
    12361 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
    12421 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
    12481 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
    12541 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
    12601 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
    12661 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccac
//
