LOCUS       sequence-370169-        9676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Universal backbone for CRISPR/Cas9 sgRNA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9676)
  AUTHORS   Heo SJ, Enriquez LD, Federman S, Chang AY, Mace R, Shevade K, Nguyen
            P, Litterman AJ, Shafer S, Przybyla L, Chow ED
  TITLE     Compact CRISPR genetic screens enabled by improved guide RNA library
            cloning.
  JOURNAL   Genome Biol. 2024 Jan 19;25(1):25. doi: 10.1186/s13059-023-03132-3.
  PUBMED    38243310
REFERENCE   2  (bases 1 to 9676)
  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: "2024-01-19"; volume: "25(1):25. doi"; pages: " 
            10.1186/s13059-023-03132-3"
COMMENT     Sequence Label: pLGR002 LGR Cas9 guide vector
FEATURES             Location/Qualifiers
     source          1..9676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        616..814
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             832..1012
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1059..1184
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1681..1914
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2099..2143
                     /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    2441..2558
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(2819..2852)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2856..2876
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     promoter        3787..4965
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4018..4956
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4913..4933
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4977..5573
                     /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(4977..4996)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5473..5493
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             5583..5636
                     /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             5643..5669
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             5685..5711
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     regulatory      5733..5742
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5739..6446
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKL"
     protein_bind    complement(6473..6506)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6562..7150
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6615..6635)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7033..7044)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7153..7169)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7154..7170)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7679..7859
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7921..8509)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8001..8020)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8680..9471,9472..9540))
                     /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     9303..9322
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9541..9645)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc
      841 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      901 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      961 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     1021 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     1081 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     1141 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     1201 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     1261 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     1321 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     1381 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     1441 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     1501 gtaagaccac cgcacagcaa gcggccggcc gctgatcttc agacctggag gaggagatat
     1561 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     1621 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     1681 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     1741 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     1801 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     1861 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     1921 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     1981 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     2041 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     2101 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     2161 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     2221 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     2281 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
     2341 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg
     2401 tgcgccaatt ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
     2461 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     2521 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
     2581 gagatccagt ttggttagta ccgggcccgc cctgcaggtc tagagatccg acgcgccatc
     2641 tctaggcccg cgccggcccc ctcgcacgga cttgtgggag aagctcggct actcccctgc
     2701 cccggttaat ttgcatataa tatttcctag taactataga ggcttaatgt gcgataaaag
     2761 acagataatc tgttcttttt aatactagct acattttaca tgataggctt ggatttctat
     2821 aacttcgtat agcatacatt atacgaagtt ataaacagca caaaaggaaa ctcaccctaa
     2881 ctgtaaagta attgtgtgtt ttgagactat aagtatccct tggagaacca ccttgttggc
     2941 tgcgactcgg cggagtccgt ttaagagcta agctggaaac agcatagcaa gtttaaataa
     3001 ggctagtccg ttatcaactt ggccgcttta aggccggtcc tagcaaggcc aagtggcacc
     3061 gagtcggtgc tttttttctc gagtactagg atccattagg cggccgccgg gaggtggtcc
     3121 ctgcagttac gccaatgata acccccgcca gaaaaatctt agtagccttc cctttttgtt
     3181 ttccgtgccc caactcggcg gattgactcg gccccttccg gaaacacccg aatcaacttc
     3241 tagtcaaatt attgttcacg ccgcaatgac ccacccctgg cccgcgtctg tggaactgac
     3301 ccctggtgta caggagagtt cgctgctgaa agtggtccca aaggggtact agtttttaag
     3361 ctcccaactc cccctccccc agcgtctgga ggattccaca ccctcgcacc gcaggggcga
     3421 ggaagtgggc ggagtccggt tttggcgcca gccgctgagg ctgccaagca gaaaagccac
     3481 cgctgaggag actccggtca ctgtcctcgc cccgcctccc ccttccctcc ccttggggac
     3541 caccgggcgc cacgccgcga acggtaagtg ccgcggtcgt cggcgcctcc gccctccccc
     3601 tagggcccca attcccagcg ggcgcggcgc gcggcccctc cccccgccgg gcgcgcgccc
     3661 gctgccccgc ccttcgtggc cgcccggcgt gggcggtgcc acccctcccc ccggcggccc
     3721 ccctgcagcc gcgcgcagct cccggctccc tcccccttcg gatgtggctt gagctgtagg
     3781 cgcggagatg catgtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     3841 ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg
     3901 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     3961 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta
     4021 agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct
     4081 tgaattactt ccacctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt
     4141 gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag
     4201 gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc
     4261 gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt
     4321 ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat ttcggttttt
     4381 ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc
     4441 ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg
     4501 gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg
     4561 gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagctcaaaa
     4621 tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc
     4681 tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac
     4741 ctcgattagt tctcgcgctt ttggagtacg tcgtctttag gttgggggga ggggttttat
     4801 gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg
     4861 atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct
     4921 cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgagctag cccaccatga
     4981 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca
     5041 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc
     5101 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     5161 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     5221 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     5281 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     5341 cgtggttcct ggccaccgtc ggcgtgtcgc ccgaccacca gggcaagggt ctgggcagcg
     5401 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     5461 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     5521 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gccggatcgg
     5581 gagagggcag aggaagtctg ctaacatgcg gtgacgtcga ggagaatcct ggcccaccgg
     5641 tccctgccgc taaacgggtc aaactcgacg gcggttctgg aggtcctgct gcaaagcgcg
     5701 ttaagttgga tggagggtca gggggagcca ccgccaccat ggtgtctaag ggcgaagagc
     5761 tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtggac aaccatcact
     5821 tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca
     5881 aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctact agcttcctct
     5941 acggcagcaa gaccttcatc aaccacaccc agggcatccc cgacttcttc aagcagtcct
     6001 tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg
     6061 ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg
     6121 tgaacttcac atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggccttca
     6181 ccgagaccct gtaccccgct gacggcggcc tggaaggcag aaacgacatg gccctgaagc
     6241 tcgtgggcgg gagccatctg atcgcaaacg ccaagaccac atatagatcc aagaaacccg
     6301 ctaagaacct caagatgcct ggcgtctact atgtggacta cagactggaa agaatcaagg
     6361 aggccaacaa cgagacctac gtcgagcagc acgaggtggc agtggccaga tactgcgacc
     6421 tccctagcaa actggggcac aagctttaat taagaattcg tcgagggacc taataacttc
     6481 gtatagcata cattatacga agttatacat gtttaagggt tccggttcca ctaggtacaa
     6541 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     6601 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     6661 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     6721 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     6781 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     6841 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     6901 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     6961 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     7021 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     7081 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     7141 gcctccccgc atcgataccg tcgacctcga tcgagaccta gaaaaacatg gagcaatcac
     7201 aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga
     7261 ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc
     7321 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     7381 acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact tccctgattg
     7441 gcagaactac acaccagggc cagggatcag atatccactg acctttggat ggtgctacaa
     7501 gctagtacca gttgagcaag agaaggtaga agaagccaat gaaggagaga acacccgctt
     7561 gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat tagagtggag
     7621 gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg actgtactgg
     7681 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     7741 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     7801 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     7861 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7921 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7981 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     8041 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     8101 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     8161 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     8221 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     8281 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     8341 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     8401 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     8461 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     8521 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     8581 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     8641 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     8701 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     8761 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     8821 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     8881 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8941 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     9001 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     9061 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     9121 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     9181 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     9241 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     9301 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     9361 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     9421 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     9481 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     9541 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     9601 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     9661 agtgccacct gacgtc
//
