LOCUS       sequence-212967-       12854 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing Lentiviral vector expressing Lentiviral
            vector expressing dCas9VP64 under the control of the long human EF1a
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12854)
  AUTHORS   Chong ZS, Ohnishi S, Yusa K, Wright GJ
  TITLE     Pooled extracellular receptor-ligand interaction screening using 
            CRISPR activation.
  JOURNAL   Genome Biol. 2018 Nov 26;19(1):205. doi: 10.1186/s13059-018-1581-3.
  PUBMED    30477585
REFERENCE   2  (bases 1 to 12854)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genome 
            Biol."; date: "2018-11-26"; volume: "19(1):205. doi"; pages: " 
            10.1186/s13059-018-1581-3"
COMMENT     Sequence Label: pKLV2-EF1adCas9VP64T2ABsd-W
FEATURES             Location/Qualifiers
     source          1..12854
                     /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             4592..4639
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(4640..4663,4664..4666,4667..7222,7223..7225,
                     7226..8740)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             8795..8815
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8840..8989
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             9011..9064
                     /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             9065..9463
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(9529..9549)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9947..9958)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10410..10427)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10416..10640
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(10666..10686)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(10666..10684)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10705..10721)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10705..10721)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10718..10740)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10729..10745
                     /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(10753..10759,10760..10777,10778..10783))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10798..10819
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10936..10953)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11107..11695)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11187..11206)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11866..12657,12658..12726))
                     /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     12489..12508
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12727..12831)
                     /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 cgcgcccgta cggccaccat gaaaaggccg gcggccacga aaaaggccgg
     4621 ccaggcaaaa aagaaaaagg acaagaagta cagcatcggc ctggccatcg gcaccaactc
     4681 tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct
     4741 gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag
     4801 cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg
     4861 gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga
     4921 cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg
     4981 gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat
     5041 ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta
     5101 tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa
     5161 ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct
     5221 gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag
     5281 actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa
     5341 tggcctgttc ggcaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa
     5401 cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct
     5461 ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa
     5521 cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc
     5581 ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct
     5641 gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag
     5701 caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt
     5761 catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag
     5821 agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca
     5881 cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga
     5941 caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct
     6001 ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc
     6061 ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat
     6121 gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta
     6181 cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag
     6241 aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac
     6301 caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt
     6361 cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca
     6421 cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat
     6481 tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg
     6541 gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag
     6601 atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc
     6661 cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca
     6721 gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg
     6781 ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa
     6841 gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa
     6901 gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa
     6961 gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat
     7021 cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta
     7081 cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga
     7141 ctacgatgtg gaccacatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa
     7201 ggtgctgacc agaagcgaca aggcccgggg caagagcgac aacgtgccct ccgaagaggt
     7261 cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag
     7321 aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg
     7381 cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct
     7441 ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt
     7501 gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt
     7561 gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac
     7621 cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt
     7681 gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa
     7741 gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg
     7801 cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga
     7861 taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt
     7921 gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa
     7981 cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga
     8041 cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa
     8101 gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga
     8161 gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat
     8221 catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc
     8281 ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt
     8341 cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa
     8401 acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga
     8461 gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa
     8521 caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct
     8581 gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag
     8641 gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct
     8701 gtacgagaca cggatcgacc tgtctcagct gggaggcgac agcgctggag gaggtggaag
     8761 cggaggagga ggaagcggag gaggaggtag cggacctaag aaaaagagga aggtggcggc
     8821 cgctggatcc ggacgggctg acgcattgga cgattttgat ctggatatgc tgggaagtga
     8881 cgccctcgat gattttgacc ttgacatgct tggttcggat gcccttgatg actttgacct
     8941 cgacatgctc ggcagtgacg cccttgatga tttcgacctg gacatgctga ttaactgtac
     9001 aggcagtgga gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg
     9061 cccaatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag caacggctac
     9121 aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc tctctagcga
     9181 cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt gtgcagaact
     9241 cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat
     9301 cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg tgcttctcga
     9361 tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga cggcagttgg
     9421 gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taagaattcg cggccgctca
     9481 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     9541 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     9601 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     9661 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     9721 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     9781 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     9841 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     9901 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     9961 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
    10021 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcaggtac ctttaagacc
    10081 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
    10141 agggctaatt cactcccaaa gaagtcaaga tctgcttttt gcctgtactg ggtctctctg
    10201 gttagaccag agtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
    10261 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
    10321 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
    10381 atctctagca gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
    10441 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
    10501 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
    10561 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gtcaatagca ggcatgctgg
    10621 ggatgcggtg ggctctatgg gcggccgtta attaagcttt tgttcccttt agtgagggtt
    10681 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
    10741 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
    10801 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
    10861 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
    10921 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
    10981 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
    11041 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
    11101 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
    11161 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
    11221 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
    11281 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
    11341 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
    11401 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
    11461 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
    11521 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
    11581 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
    11641 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
    11701 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
    11761 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
    11821 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
    11881 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
    11941 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
    12001 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
    12061 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
    12121 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
    12181 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
    12241 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
    12301 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
    12361 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
    12421 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
    12481 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
    12541 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
    12601 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
    12661 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
    12721 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
    12781 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
    12841 ccgaaaagtg ccac
//
