LOCUS       sequence-169872-       10952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of guide RNA (mup53_Ex5) with fluorescent GFP 
            reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10952)
  AUTHORS   Aubrey BJ, Kelly GL, Kueh AJ, Brennan MS, O'Connor L, Milla L, 
            Wilcox S, Tai L, Strasser A, Herold MJ
  TITLE     An inducible lentiviral guide RNA platform enables the 
            identification of tumor-essential genes and tumor-promoting 
            mutations in vivo.
  JOURNAL   Cell Rep. 2015 Mar 3;10(8):1422-32. doi: 
            10.1016/j.celrep.2015.02.002. Epub 2015 Feb 26.
  PUBMED    25732831
REFERENCE   2  (bases 1 to 10952)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2015.02.002"; journalName: "Cell Rep"; date: 
            "2015-03-3- 3"; volume: "10"; issue: "8"; pages: "1422-32"
COMMENT     Sequence Label: FgH1tUTG_mup53_Ex5
FEATURES             Location/Qualifiers
     source          1..10952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    43..267
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     256..273
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             299..479
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    989..1577
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1095..1106
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1504..1524
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1617..2330,2331..2333,2334..2336))
                     /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             complement(join(1620..2330,2331..2333,2334..2336))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1662..1683)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2011..2030
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2270..2291
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(2337..2390)
                     /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             complement(2400..3020)
                     /codon_start=1
                     /gene="synthetic gene"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     promoter        complement(3058..4269)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(3202..3221)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     misc_RNA        complement(4305..4380)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    4406..4424
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(4469..4488)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_feature    4679..4796
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(5094..5138)
                     /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    5323..5556
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    6049..6174
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6221..6401
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(6419..6617)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6444..6464)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6619..6998
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     7170..7189
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7264..7368
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7369..7437,7438..8229)
                     /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     complement(7587..7606)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8400..8988
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8889..8908
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9142..9159
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9276..9297
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9312..9317,9318..9335,9336..9342)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9350..9366
                     /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)."
     primer_bind     9355..9377
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9374..9390
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9374..9390
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    9439..9560
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9450..9469)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     9504..9523
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(9690..10064)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(10083..10130)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(10178..10507)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10192..10327
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10246..10265)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     10492..10512
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(10521..10949)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10631..10652)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10843..10862
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gataccccct agagccccag ctggttcttt ccgcctcaga agccatagag cccaccgcat
       61 ccccagcatg cctgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac
      121 cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg
      181 aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa
      241 cagatggctg gcaactagaa ggcacagtcg aggctgatca gcgggtttaa acgggccctg
      301 ctagagattt tccacactga ctaaaagggt ctgagggatc tctagttacc agagtcacac
      361 aacagacggg cacacactac ttgaagcact caaggcaagc tttattgagg cttaagcagt
      421 gggttcccta gttagccaga gagctcccag gctcagatct ggtctaacca gagagaccca
      481 gtacagtccg gatgcagctc tcgggccatg tgatgaaatg ctaggcggct gtcaaacctc
      541 cactctaata cttctctctc cgggtcatcc atcccatgca ggctcacagg gtgtaacaag
      601 cgggtgttct ctccttcatt ggcttcttct accttctctt gctcaactgg tactagcttg
      661 tagcaccatc caaaggtcag tggatatctg atccctggcc ctggtgtgta gttctgccaa
      721 tcagggaagt agccttgtgt gtggtagatc cacagatcaa ggatatcttg tcttcgttgg
      781 gagtgaatta gcccttccag tccccccttt tcttttaaaa agtggctaag atctacagct
      841 gccttgtaag tcattggtct taaaggtacc tgaggtgtga ctggaaaacc cacctcctcc
      901 tcctcttgtg cttctagcca ggcacaatca gcattggtag ctgctgtatt gctacttgtg
      961 attgctccat gtttttctag gtctcgatgc ggggaggcgg cccaaaggga gatccgactc
     1021 gtctgagggc gaaggcgaag acgcggaaga ggccgcagag ccggcagcag gccgcgggaa
     1081 ggaaggtccg ctggattgag ggccgaaggg acgtagcaga aggacgtccc gcgcagaatc
     1141 caggtggcaa cacaggcgag cagccaagga aaggacgatg atttccccga caacaccacg
     1201 gaattgtcag tgcccaacag ccgagcccct gtccagcagc gggcaaggca ggcggcgatg
     1261 agttccgccg tggcaatagg gagggggaaa gcgaaagtcc cggaaaggag ctgacaggtg
     1321 gtggcaatgc cccaaccagt gggggttgcg tcagcaaaca cagtgcacac cacgccacgt
     1381 tgcctgacaa cgggccacaa ctcctcataa agagacagca accaggattt atacaaggag
     1441 gagaaaatga aagccatacg ggaagcaata gcatgataca aaggcattaa agcagcgtat
     1501 ccacatagcg taaaaggagc aacatagtta agaataccag tcaatctttc acaaattttg
     1561 taatccagag gttgattatc gataagcttg atatcgaatt gtagatcgcg gccgctttac
     1621 ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc
     1681 atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actgggtgct caggtagtgg
     1741 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg
     1801 agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc
     1861 ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc cagcttgtgc
     1921 cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg
     1981 gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag
     2041 atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc
     2101 atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt cacgagggtg
     2161 ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag
     2221 gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc
     2281 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct caccattggg
     2341 ccaggattct cctcgacgtc accgcatgtt agcagacttc ctctgccctc accggaccca
     2401 gagccagact cacatttcag ctgcttttcc aggccacaga tgatcagttc caggccaaaa
     2461 aggaatgcag gctcagcccc ttggtggtca aacaactcaa tagcttgccg cagcagtggt
     2521 ggcatggagt cagtggtggg ggtctccctc tcctccttgg caacttggtg ctcctggtcc
     2581 tccagcacac agcccaaggt gaagtggccc acagcagaga gggcatacaa ggcattttcc
     2641 agggagaagc cttgttggca caggaatgcc agctggttct ccagggtctc atactgcttc
     2701 tctgtaggcc tggtgcccaa gtgaactttg gcaccatccc ggtgggagag caaggcacat
     2761 ctgaagctct tggcattgtt cctcaggaag tcctgccagg actccccttc cagagggcag
     2821 aagtgggtgt ggtgcctgtc caacatctca atggccaggg catcaagcag agccctcttg
     2881 ttcttcacat gccagtacag ggtaggctgc tccacaccca gcttctgggc cagcttcctg
     2941 gtggtcaagc cctcaatgcc aacttcattc aggagctcca aggcagagtt gatgactttg
     3001 ctcttgtcca gtctggacat ggtggatcct ctagagtcga cctgcagccc aagcttcgtc
     3061 taacaaaaaa gccaaaaacg gccagaattt agcggacaat ttactagtct aacactgaaa
     3121 attacatatt gacccaaatg attacatttc aaaaggtgcc taaaaaactt cacaaaacac
     3181 actcgccaac cccgagcgca tagttcaaaa ccggagcttc agctacttaa gaagataggt
     3241 acataaaacc gaccaaagaa actgacgcct cacttatccc tcccctcacc agaggtccgg
     3301 cgcctgtcga ttcaggagag cctaccctag gcccgaaccc tgcgtcctgc gacggagaaa
     3361 agcctaccgc acacctaccg gcaggtggcc ccaccctgca ttataagcca acagaacggg
     3421 tgacgtcacg acacgacgag ggcgcgcgct cccaaaggta cgggtgcact gcccaacggc
     3481 accgccataa ctgccgcccc cgcaacagac gacaaaccga gttctccagt cagtgacaaa
     3541 cttcacgtca gggtccccag atggtgcccc agcccatctc acccgaataa gagctttccc
     3601 gcattagcga aggcctcaag accttgggtt cttgccgccc accatgcccc ccaccttgtt
     3661 tcaacgacct cacagcccgc ctcacaagcg tcttccattc aagactcggg aacagccgcc
     3721 attttgctgc gctcccccca acccccagtt cagggcaacc ttgctcgcgg acccagacta
     3781 cagcccttgg cggtctctcc acacgcttcc gtcccaccga gcggcccggc ggccacgaaa
     3841 gccccggcca gcccagcagc ccgctactca ccaagtgacg atcacagcga tccacaaaca
     3901 agaaccgcga cccaaatccc ggctgcgacg gaactagctg tgccacaccc ggcgcgtcct
     3961 tatataatca tcggcgttca ccgccccacg gagatccctc cgcagaatcg ccgagaaggg
     4021 actacttttc ctcgcctgtt ccgctctctg gaaagaaaac cagtgcccta gagtcaccca
     4081 agtcccgtcc taaaatgtcc ttctgctgat actggggttc taaggccgag tcttatgagc
     4141 agcgggccgc tgtcctgagc gtccgggcgg aaggatcagg acgctcgctg cgcccttcgt
     4201 ctgacgtggc agcgctcgcc gtgaggaggg gggcgcccgc gggaggcgcc aaaacccggc
     4261 gcggaggcca gatctggagc cgacacgggt taattaaaaa aaaagcaccg actcggtgcc
     4321 actttttcaa gttgataacg gactagcctt attttaactt gctatttcta gctctaaaac
     4381 tcaccatcgg agcagcgctc gggaatctct atcactgata gggacttata agattcccaa
     4441 atccaaagac atttcacgtt tatggtgatt tcccagaaca catagcgaca tgcaaatatt
     4501 gcagggcgcc actcccctgt ccctcacagc catcttcctg ccagggcgca cgcgcgctgg
     4561 gtgttcccgc ctagtgacac tgggcccgcg attccttgga gcgggttgat gacgtcagcg
     4621 ttcgaattcc atggttaatt aaccaaactg gatctctgct gtccctgtaa taaacccgaa
     4681 aattttgaat ttttgtaatt tgtttttgta attctttagt ttgtatgtct gttgctatta
     4741 tgtctactat tctttcccct gcactgtacc ccccaatccc cccttttctt ttaaaattgt
     4801 ggatgaatac tgccatttgt ctgcagaatt ggcgcacgca gtgccgatcc gttcactaat
     4861 cgaatggatc tgtctctgtc tctctctcca ccttcttctt ctattccttc gggcctgtcg
     4921 ggtcccctcg gggttgggag gtgggtctga aacgataatg gtgaatatcc ctgcctaact
     4981 ctattcacta tagaaagtac agcaaaaact attcttaaac ctaccaagcc tcctactatc
     5041 attatgaata attttatata ccacagccaa tttgttatgt taaaccaatt ccacaaactt
     5101 gcccatttat ctaattccaa taattcttgt tcattctttt cttgctggtt ttgcgattct
     5161 tcaattaagg agtgtattaa gcttgtgtaa ttgttaattt ctctgtccca ctccatccag
     5221 gtcgtgtgat tccaaatctg ttccagagat ttattactcc aactagcatt ccaaggcaca
     5281 gcagtggtgc aaatgagttt tccagagcaa ccccaaatcc ccaggagctg ttgatccttt
     5341 aggtatcttt ccacagccag gattcttgcc tggagctgct tgatgcccca gactgtgagt
     5401 tgcaacagat gctgttgcgc ctcaatagcc ctcagcaaat tgttctgctg ctgcactata
     5461 ccagacaata attgtctggc ctgtaccgtc agcgtcattg acgctgcgcc catagtgctt
     5521 cctgctgctc ccaagaaccc aaggaacaaa gctcctattc ccactgctct tttttctctc
     5581 tgcaccactc ttctctttgc cttggtgggt gctactccta atggttcaat ttttactact
     5641 ttatatttat ataattcact tctccaattg tccctcatat ctcctcctcc aggtctgaag
     5701 atcagcggcc gcttgctgtg cggtggtctt acttttgttt tgctcttcct ctatcttgtc
     5761 taaagcttcc ttggtgtctt ttatctctat cctttgatgc acacaataga gggttgctac
     5821 tgtattatat aatgatctaa gttcttctga tcctgtctga agggatggtt gtagctgtcc
     5881 cagtatttgt ctacagcctt ctgatgtttc taacaggcca ggattaactg cgaatcgttc
     5941 tagctccctg cttgcccata ctatatgttt taatttatat tttttctttc cccctggcct
     6001 taaccgaatt ttttcccatc gcgatctaat tctcccccgc ttaatactga cgctctcgca
     6061 cccatctctc tccttctagc ctccgctagt caaaattttt ggcgtactca ccagtcgccg
     6121 cccctcgcct cttgccgtgc gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc
     6181 tggtttccct ttcgctttca agtccctgtt cgggcgccac tgctagagat tttccacact
     6241 gactaaaagg gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact
     6301 acttgaagca ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca
     6361 gagagctccc aggctcagat ctggtctaac cagagagacc cagtacaggc aaaacgcgct
     6421 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
     6481 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
     6541 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
     6601 ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt
     6661 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     6721 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     6781 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     6841 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     6901 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
     6961 aattgattac tattaataac tagtcaataa tcaatgtcaa cgcgtatatc tggcccgtac
     7021 atcgcgaagc agcgcaaaac gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa
     7081 gccttgcctt gttgtagctt aaattttgct cgcgcactac tcagcgacct ccaacacaca
     7141 agcagggagc agatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     7201 ccatagggga tcgggagatc tcccgatccg tcgacgtcag gtggcacttt tcggggaaat
     7261 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     7321 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     7381 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     7441 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     7501 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     7561 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     7621 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     7681 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     7741 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     7801 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     7861 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     7921 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     7981 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     8041 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     8101 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     8161 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     8221 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     8281 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     8341 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     8401 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     8461 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     8521 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     8581 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     8641 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     8701 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     8761 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     8821 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     8881 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     8941 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     9001 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     9061 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     9121 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     9181 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     9241 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     9301 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     9361 taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcta gctagaggtc
     9421 gacggtatac agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     9481 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     9541 taagctgcaa taaacaagtt ggggtgggcg aagaactcca gcatgagatc cccgcgctgg
     9601 aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg
     9661 gcggtggaat cgaaatctcg tagcacgtgt cagtcctgct cctcggccac gaagtgcacg
     9721 cagttgccgg ccgggtcgcg cagggcgaac tcccgccccc acggctgctc gccgatctcg
     9781 gtcatggccg gcccggaggc gtcccggaag ttcgtggaca cgacctccga ccactcggcg
     9841 tacagctcgt ccaggccgcg cacccacacc caggccaggg tgttgtccgg caccacctgg
     9901 tcctggaccg cgctgatgaa cagggtcacg tcgtcccgga ccacaccggc gaagtcgtcc
     9961 tccacgaagt cccgggagaa cccgagccgg tcggtccaga actcgaccgc tccggcgacg
    10021 tcgcgcgcgg tgagcaccgg aacggcactg gtcaacttgg ccatggttta gttcctcacc
    10081 ttgtcgtatt atactatgcc gatatactat gccgatgatt aattgtcaac acgtgctgat
    10141 cagatccgaa aatggatata caagctcccg ggagcttttt gcaaaagcct aggcctccaa
    10201 aaaagcctcc tcactacttc tggaatagct cagaggcaga ggcggcctcg gcctctgcat
    10261 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
    10321 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt
    10381 gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga
    10441 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact
    10501 gacacacatt ccacagaatt aattcgcgtt aaatttttgt taaatcagct cattttttaa
    10561 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
    10621 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
    10681 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
    10741 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
    10801 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
    10861 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
    10921 cgcgcttaat gcgccgctac agggcgcgtg gg
//
