LOCUS       sequence-217731-       11059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector carrying human EF1a promoter-driven 
            dCas9VP64-T2A-MS2p65HSF1 with Blasticidine resistant marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11059)
  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 11059)
  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: pENTR-EF1adCas9VP64_T2A_MS2p65HSF1-IRESbsdpA
FEATURES             Location/Qualifiers
     source          1..11059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          231..1173
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1130..1150
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1212..1259
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(1260..1283,1284..1286,1287..3842,3843..3845,
                     3846..5360)
                     /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             5415..5435
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5460..5609
                     /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             5640..5693
                     /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"
     regulatory      5700..5709
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5709..6095
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2-N55K
                     /note="binds with high affinity to a specific stem-loop 
                     structure in the viral RNA (Lim et al., 1994)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     CDS             6150..6170
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6186..6728
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     CDS             6753..7124
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     misc_feature    join(7149..7723,7724..7726,7727..7735)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7316..7333)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7543..7562
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7736..8131
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    8135..8359
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(8380..8479)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(8591..8610)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             8602..9411
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(8669..8688)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      9501..10089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9990..10009
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      10254..10340
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      10432..10459
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     10467..10490
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    10509..10608
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(11059,1..1181)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
ORIGIN
        1 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
       61 gaggggtcgg caattgaacc ggtgcctaga gaaagtggcg cggggtaaac tgggaaagtg
      121 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
      181 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
      241 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
      301 cttccacgcc cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
      361 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
      421 tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
      481 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
      541 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
      601 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
      661 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
      721 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
      781 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
      841 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
      901 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
      961 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
     1021 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
     1081 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
     1141 acagtggttc aaagtttttt tcttccattt caggtgtcgt ggaagcttgg cgcgcccgta
     1201 cggccaccat gaaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagg
     1261 acaagaagta cagcatcggc ctggccatcg gcaccaactc tgtgggctgg gccgtgatca
     1321 ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca
     1381 gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca
     1441 cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc
     1501 tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg
     1561 aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca
     1621 tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac
     1681 tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga
     1741 tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg
     1801 acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca
     1861 acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc
     1921 tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggcaacctga
     1981 ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg
     2041 ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga
     2101 tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc
     2161 tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga
     2221 tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc
     2281 agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct
     2341 acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa
     2401 agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc
     2461 agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca
     2521 ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga
     2581 agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat
     2641 tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg
     2701 tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc
     2761 tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata
     2821 acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg
     2881 gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga
     2941 agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg
     3001 gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca
     3061 aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga
     3121 ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc
     3181 tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc
     3241 tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt
     3301 tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc
     3361 tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg
     3421 agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga
     3481 aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg
     3541 aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga
     3601 agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg
     3661 aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata
     3721 tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccacatcg
     3781 tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca
     3841 aggcccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact
     3901 actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca
     3961 aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg
     4021 tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta
     4081 agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc
     4141 tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc
     4201 accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc
     4261 ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga
     4321 tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca
     4381 tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc
     4441 tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca
     4501 ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga
     4561 caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca
     4621 gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt
     4681 ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag
     4741 agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc
     4801 tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact
     4861 ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga
     4921 agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact
     4981 atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc
     5041 acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc
     5101 tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca
     5161 tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg
     5221 ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg
     5281 tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc
     5341 tgtctcagct gggaggcgac agcgctggag gaggtggaag cggaggagga ggaagcggag
     5401 gaggaggtag cggacctaag aaaaagagga aggtggcggc cgctggatcc ggacgggctg
     5461 acgcattgga cgattttgat ctggatatgc tgggaagtga cgccctcgat gattttgacc
     5521 ttgacatgct tggttcggat gcccttgatg actttgacct cgacatgctc ggcagtgacg
     5581 cccttgatga tttcgacctg gacatgctga ttaactgtac gggcggcggg tccggaggag
     5641 agggcagagg aagtctccta acatgcggtg acgtggagga gaatcctggc ccatgtacgg
     5701 ccaccatggc ttcaaacttt actcagttcg tgctcgtgga caatggtggg acaggggatg
     5761 tgacagtggc tccttctaat ttcgctaatg gggtggcaga gtggatcagc tccaactcac
     5821 ggagccaggc ctacaaggtg acatgcagcg tcaggcagtc tagtgcccag aagagaaagt
     5881 ataccatcaa ggtggaggtc cccaaagtgg ctacccagac agtgggcgga gtcgaactgc
     5941 ctgtcgccgc ttggaggtcc tacctgaaca tggagctcac tatcccaatt ttcgctacca
     6001 attctgactg tgaactcatc gtgaaggcaa tgcaggggct cctcaaagac ggtaatccta
     6061 tcccttccgc catcgccgct aactcaggta tctacagcgc tggaggaggt ggaagcggag
     6121 gaggaggaag cggaggagga ggtagcggac ctaagaaaaa gaggaaggtg gcggccgctg
     6181 gatccccttc agggcagatc agcaaccagg ccctggctct ggcccctagc tccgctccag
     6241 tgctggccca gactatggtg ccctctagtg ctatggtgcc tctggcccag ccacctgctc
     6301 cagcccctgt gctgacccca ggaccacccc agtcactgag cgctccagtg cccaagtcta
     6361 cacaggccgg cgaggggact ctgagtgaag ctctgctgca cctgcagttc gacgctgatg
     6421 aggacctggg agctctgctg gggaacagca ccgatcccgg agtgttcaca gatctggcct
     6481 ccgtggacaa ctctgagttt cagcagctgc tgaatcaggg cgtgtccatg tctcatagta
     6541 cagccgaacc aatgctgatg gagtaccccg aagccattac ccggctggtg accggcagcc
     6601 agcggccccc cgaccccgct ccaactcccc tgggaaccag cggcctgcct aatgggctgt
     6661 ccggagatga agacttctca agcatcgctg atatggactt tagtgccctg ctgtcacaga
     6721 tttcctctag tgggcaggga ggaggtggaa gcggcttcag cgtggacacc agtgccctgc
     6781 tggacctgtt cagcccctcg gtgaccgtgc ccgacatgag cctgcctgac cttgacagca
     6841 gcctggccag tatccaagag ctcctgtctc cccaggagcc ccccaggcct cccgaggcag
     6901 agaacagcag cccggattca gggaagcagc tggtgcacta cacagcgcag ccgctgttcc
     6961 tgctggaccc cggctccgtg gacaccggga gcaacgacct gccggtgctg tttgagctgg
     7021 gagagggctc ctacttctcc gaaggggacg gcttcgccga ggaccccacc atctccctgc
     7081 tgacaggctc ggagcctccc aaagccaagg accccactgt ctcctgtaca tgagaattcc
     7141 gccccccccc cntctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
     7201 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     7261 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     7321 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     7381 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     7441 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     7501 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     7561 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     7621 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     7681 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatggc
     7741 caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag
     7801 catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat
     7861 cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct
     7921 gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga
     7981 gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc
     8041 tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg ggattcgtga
     8101 attgctgccc tctggttatg tgtgggaggg ccgactgtgc cttctagttg ccagccatct
     8161 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     8221 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     8281 ggtggggtgg ggcaggacag caagggggag gattgggaag tcaatagcag gcatgctggg
     8341 gatgcggtgg gctctatgga ctagtctcga gatatctaga cccagctttc ttgtacaaag
     8401 ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
     8461 aaaataaaat cattatttgc catccagctg cagctctggc ccgtgtctca aaatctctga
     8521 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     8581 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     8641 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     8701 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     8761 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     8821 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     8881 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     8941 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     9001 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     9061 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     9121 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     9181 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     9241 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     9301 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     9361 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     9421 gttaattggt tgtaacatta ttcagattgg gccccgttcc actgagcgtc agaccccgta
     9481 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     9541 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     9601 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     9661 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     9721 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     9781 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     9841 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     9901 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     9961 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
    10021 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
    10081 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
    10141 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgctagc
    10201 atggatctcg gggacgtcta actactaagc gagagtaggg aactgccagg catcaaataa
    10261 aacgaaaggc tcagtcggaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
    10321 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgtgaag caacggcccg
    10381 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaactaag cagaaggcca
    10441 tcctgacgga tggccttttt gcgtttctac aaactcttcc tgttagttag ttacttaagc
    10501 tcgggcccca aataatgatt ttattttgac tgatagtgac ctgttcgttg caacaaattg
    10561 ataagcaatg cttttttata atgccaactt tgtacaaaaa agcaggctgg cgccggaacc
    10621 aattcagtcg actggatcca cgcgtgaggg gcgcggaagc cggggcggag gaggagccaa
    10681 ccccgggctg ggctgagacc cgcagaggaa gacgctctag ggatttgtcc cggactagcg
    10741 agatggcaag gctgaggacg ggaggctgat tgagaggcga aggtacaccc taatctcaat
    10801 acaacctttg gagctaagcc agcaatggta gagggaagat tctgcacgtc ccttccaggc
    10861 ggcctccccg tcaccacccc ccccaacccg ccccgaccgg agctgagagt aattcataca
    10921 aaaggactcg cccctgcctt ggggaatccc agggaccgtc gttaaactcc cactaacgta
    10981 gaacccagag atcgctgcgt tcccgccccc tcacccgccc gctctcgtca tcactgaggt
    11041 ggagaagagc atgcgtgag
//
