LOCUS       sequence-217738-        9757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR-a reporter assay lentiviral vector (empty gRNA control).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9757)
  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 9757)
  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-U6gRNASAM(gEmpty)-TREGFP-PGKpuroBFP-W
FEATURES             Location/Qualifiers
     source          1..9757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        285..303
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        355..373
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_feature    441..558
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        619..933
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    627..645
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    663..681
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    698..716
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    734..752
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    770..788
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    805..823
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     906..930
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(949..951,952..954,955..1668)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(949..951,952..954,955..1665)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(994..1015)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1255..1274)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1602..1623
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        1684..2183
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1719..1742)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2116..2135
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2223..2819
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2223..2242)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2719..2739
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2838..2891
                     /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             2892..3593
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(3653..3673)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4071..4082)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4534..4551)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4540..4764
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4790..4810)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4790..4808)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4829..4845)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4829..4845)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4842..4864)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4853..4869
                     /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(4877..4883,4884..4901,4902..4907))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4922..4943
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5060..5077)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5231..5819)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5311..5330)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5990..6781,6782..6850))
                     /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     6613..6632
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6851..6955)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6981..7436)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7118..7139)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7330..7349
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7563..7585
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7577..7594
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7578..7594
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7604..7623
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7604..7622
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        7938..8136
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             8154..8334
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8381..8506
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9005..9238
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9423..9467
                     /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"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg ggtcttcgag aagacacgtt taagagctag gccaacatga ggatcaccca
      301 tgtctgcagg gcctagcaag tttaaataag gctagtccgt tatcaacttg gccaacatga
      361 ggatcaccca tgtctgcagg gccaagtggc accgagtcgg tgcttttttt ctgcagacaa
      421 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
      481 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
      541 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttggttagta
      601 ccgggcccta cgcgtctcga gtttactccc tatcagtgat agagaacgta tgtcgagttt
      661 actccctatc agtgatagag aacgatgtcg agtttactcc ctatcagtga tagagaacgt
      721 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt
      781 gatagagaac gtatgtcgag tttatcccta tcagtgatag agaacgtatg tcgagtttac
      841 tccctatcag tgatagagaa cgtatgtcga ggtaggcgtg tacggtggga ggcctatata
      901 agcagagctc gtttagtgaa ccgtcagatc gcctggagaa ttcccaccat ggtgagcaag
      961 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1021 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1081 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1141 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     1201 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     1261 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1321 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1381 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     1441 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1501 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1561 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1621 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaagg atccaattct
     1681 accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc
     1741 tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta
     1801 ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag
     1861 tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag
     1921 cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc
     1981 tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa
     2041 ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt
     2101 cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct
     2161 cttcctcatc tccgggcctt tcgacctgca tccatctaga tctcgagcag ctgaagctta
     2221 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     2281 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     2341 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     2401 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     2461 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     2521 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     2581 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     2641 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     2701 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     2761 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgccg
     2821 gcggcgggtc cggaggagag ggcagaggaa gtctcctaac atgcggtgac gtggaggaga
     2881 atcctggccc aatgagcgag ctgattaagg agaacatgca catgaagctg tacatggagg
     2941 gcaccgtgga caaccatcac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg
     3001 gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc gccttcgaca
     3061 tcctggctac tagcttcctc tacggcagca agaccttcat caaccacacc cagggcatcc
     3121 ccgacttctt caagcagtcc ttccctgagg gcttcacatg ggagagagtc accacatacg
     3181 aggacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct
     3241 acaacgtcaa gatcagaggg gtgaacttca catccaacgg ccctgtgatg cagaagaaaa
     3301 cactcggctg ggaggccttc accgagacgc tgtaccccgc tgacggcggc ctggaaggca
     3361 gaaacgacat ggccctgaag ctcgtgggcg ggagccatct gatcgcaaac atcaagacca
     3421 catatagatc caagaaaccc gctaagaacc tcaagatgcc tggcgtctac tatgtggact
     3481 acagactgga aagaatcaag gaggccaaca acgagaccta cgtcgagcag cacgaggtgg
     3541 cagtggccag atactgcgac ctccctagca aactggggca caagcttaat tgagcggccg
     3601 ctcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3661 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3721 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3781 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3841 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3901 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3961 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     4021 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     4081 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtctac
     4141 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcag gtacctttaa
     4201 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     4261 tggaagggct aattcactcc caaagaagtc aagatctgct ttttgcctgt actgggtctc
     4321 tctggttaga ccagagtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     4381 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     4441 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     4501 gaaaatctct agcagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     4561 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     4621 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     4681 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagtcaat agcaggcatg
     4741 ctggggatgc ggtgggctct atgggcggcc gttaattaag cttttgttcc ctttagtgag
     4801 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4861 cgctcacaat tccacacaac atacgagccg ggagcataaa gtgtaaagcc tggggtgcct
     4921 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4981 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5041 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5101 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5161 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5221 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5281 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5341 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5401 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5461 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5521 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5581 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5641 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5701 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5761 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5821 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5881 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5941 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6001 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6061 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6121 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6181 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6241 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6301 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6361 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6421 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6481 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6541 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6601 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6661 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6721 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6781 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6841 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6901 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6961 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     7021 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     7081 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     7141 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     7201 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     7261 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     7321 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     7381 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
     7441 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     7501 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     7561 ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata
     7621 gggcgaattg actagtacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     7681 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     7741 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     7801 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     7861 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     7921 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     7981 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     8041 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     8101 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
     8161 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     8221 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     8281 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     8341 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     8401 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     8461 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     8521 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     8581 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     8641 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     8701 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     8761 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     8821 aagtaagacc accgcacagc aagcggccgg ccgcgctgat cttcagacct ggaggaggag
     8881 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     8941 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     9001 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     9061 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     9121 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     9181 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     9241 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     9301 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     9361 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     9421 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     9481 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     9541 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     9601 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     9661 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac
     9721 tgcgtgcgcc aattctgcag ccaaggtcgg gcaggaa
//
