LOCUS       sequence-314272-       10615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses S. pyogenes CRISPR chimeric RNA element with customizable 
            sgRNA from U6 promoter and hygromycin resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10615)
  AUTHORS   Gstalder C, Liu D, Miao D, Lutterbach B, DeVine AL, Lin C, Shettigar
            M, Pancholi P, Buchbinder EI, Carter SL, Manos MP, Rojas-Rudilla V, 
            Brennick R, Gjini E, Chen PH, Lako A, Rodig S, Yoon CH, Freeman GJ, 
            Barbie DA, Hodi FS, Miles W, Van Allen EM, Haq R
  TITLE     Inactivation of Fbxw7 impairs dsRNA sensing and confers resistance 
            to PD-1 blockade.
  JOURNAL   Cancer Discov. 2020 May 5. pii: 2159-8290.CD-19-1416. doi: 
            10.1158/2159-8290.CD-19-1416.
  PUBMED    32371478
REFERENCE   2  (bases 1 to 10615)
  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: "Cancer 
            Discov. 2020 May 5. pii: 2159-8290.CD-19-1416. doi: 
            10.1158/2159-8290.CD-19-1416."
COMMENT     Sequence Label: LentiGuide-hygro
FEATURES             Location/Qualifiers
     source          1..10615
                     /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        2131..2206
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2262..2379
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2518..3696
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2749..3687
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3644..3664
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3715..4740
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3715..4740
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     misc_feature    4756..5344
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4809..4829)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5227..5238)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5416..5649
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5727..5848
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5764..5783)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5818..5837
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5888..6023
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6043..6062)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6044..6062)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6072..6089)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6072..6088)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6081..6103)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6230..6685
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6317..6336)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6527..6548
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6711..6815
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6816..6884,6885..7676)
                     /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(7034..7053)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7847..8435
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8336..8355
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8589..8606
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8723..8744
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8759..8764,8765..8782,8783..8789)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8797..8813
                     /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     8802..8824
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8821..8837
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8821..8837
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8856..8876
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8858..8876
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8904..9130
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             9131..9311
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9358..9483
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9976..10209
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10394..10438
                     /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 gagacggttg taaatgagca cacaaaatac acatgctaaa atattatatt
      301 ctatgacctt tataaaatca accaaaatct tctttttaat aactttagta tcaataatta
      361 gaatttttat gttccttttt gcaaactttt aataaaaatg agcaaaataa aaaaacgcta
      421 gttttagtaa ctcgcgttgt tttcttcacc tttaataata gctactccac cacttgttcc
      481 taagcggtca gctcctgctt caatcatttt ttgagcatct tcaaatgttc taactccacc
      541 agctgcttta actaaagcat tgtctttaac aactgacttc attagtttaa catcttcaaa
      601 tgttgcacct gattttgaaa atcctgttga tgttttaaca aattctaatc cagcttcaac
      661 agctatttca caagctttca tgatttcttc ttttgttaat aaacaatttt ccataataca
      721 tttaacaaca tgtgatccag ctgctttttt tacagctttc atgtcttcta aaactaattc
      781 ataatttttg tcttttaatg caccaatatt taataccata tcaatttctg ttgcaccatc
      841 tttaattgct tcagaaactt cgaatgcttt tgtagctgtt gtgcatgcac ctagaggaaa
      901 acctacaaca tttgttattc ctacatttgt gccttttaat aattctttac aatagcttgt
      961 tcaatatgaa ttaacacaaa ctgttgcaaa atcaaattca attgcttcat cacataattg
     1021 tttaatttca gctttcgtag catcttgttt taataatgtg tgatctatat atttgtttag
     1081 tttcattttt tctcctatat attcattttt aattttaatt ctttaataat ttcgtctact
     1141 ttaactttag cgttttgaac agattcacca acacctataa aataaatttt tagtttaggt
     1201 tcagttccac ttgggcgaac agcaaatcat gacttatctt ctaaataaaa ttttagtaag
     1261 tcttgtcctg gcatattata cattccatcg atgtagtctt caacattaac aactttaagt
     1321 ccagcaattt gagttaaggg tgttgctctc aatgatttca ttaatggttc aatttttaat
     1381 ttcttttctt ctggtttaaa attcaagttt aaagtgaaag tgtaatatgc acccatttct
     1441 ttaaataaat cttctaaata gtctactaat gttttatttt gttttttata aaatcaagca
     1501 gcctctgcta ttaatataga agcttgtatt ccatctttat ctctagctga gtcatcaatt
     1561 acatatccat aactttcttc ataagcaaaa acaaaattta atccgttatc ttcttcttta
     1621 gcaatttctc tacccattca tttaaatcca gttaaagttt ttacaatatt aactccatat
     1681 ttttcatgag cgattctatc acccaaatca cttgttacaa aacttgaata tagagccgga
     1741 ttttttggaa tgctatttaa gcgttttaga tttgataatt ttcaatcaat taaaattggt
     1801 cctgtttgat ttccatctaa tcttacaaaa tgaccatcat gttttattgc cattccaaat
     1861 ctgtcagcat ctgggtcatt cataataata atatctgcat catgtttaat accatattca
     1921 agcggtattt ttcatgcagg atcaaattct ggatttggat ttacaacatt tttaaatgtt
     1981 tcatcttcaa atgcatgctc ttcaacctca ataacgttat atcctgattc acgtaatatt
     2041 tttggggtaa atttagttcc tgttccatta actgcgctaa aaataatttt taaatctttt
     2101 ttagcttctt gctctttttt gtacgtctct gttttagagc tagaaatagc aagttaaaat
     2161 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttaagcttgg
     2221 cgtaactaga tcttgagaca aatggcagta ttcatccaca attttaaaag aaaagggggg
     2281 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
     2341 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
     2401 agagatccac tttggcgccg gctcgagggg gcccgggtgc aaagatggat aaagttttaa
     2461 acagagagga atctttgcag ctaatggacc ttctaggtct tgaaaggagt gggaattggc
     2521 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
     2581 ggggtcggca attgatccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
     2641 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     2701 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg
     2761 tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact
     2821 tccacctggc tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag
     2881 agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct
     2941 gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc
     3001 gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa
     3061 gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg
     3121 ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg
     3181 cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc
     3241 ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt
     3301 gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg
     3361 cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc
     3421 tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag
     3481 ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag
     3541 tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc
     3601 tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg
     3661 gttcaaagtt tttttcttcc atttcaggtg tcgtgacgta cggccacctt cgaaatgaaa
     3721 aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgta
     3781 tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga
     3841 gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat
     3901 gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa
     3961 tttagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac
     4021 ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc
     4081 gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt
     4141 caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg
     4201 caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg
     4261 ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac
     4321 aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc
     4381 ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg
     4441 gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc
     4501 cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat
     4561 ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg
     4621 actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta
     4681 gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag
     4741 acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     4801 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     4861 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     4921 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     4981 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     5041 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     5101 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     5161 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     5221 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     5281 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     5341 ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     5401 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg
     5461 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     5521 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     5581 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     5641 tctctagcag tacgtatagt agttcatgtc atcttattat tcagtattta taacttgcaa
     5701 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat
     5761 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5821 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac
     5881 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     5941 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     6001 gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat
     6061 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     6121 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     6181 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     6241 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     6301 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     6361 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     6421 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     6481 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     6541 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     6601 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     6661 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     6721 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     6781 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     6841 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     6901 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     6961 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     7021 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     7081 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     7141 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     7201 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     7261 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     7321 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     7381 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     7441 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     7501 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     7561 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     7621 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     7681 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     7741 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     7801 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     7861 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     7921 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     7981 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     8041 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     8101 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     8161 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     8221 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     8281 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     8341 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     8401 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     8461 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     8521 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     8581 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     8641 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     8701 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     8761 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     8821 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
     8881 aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat
     8941 ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt
     9001 ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg
     9061 gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg
     9121 atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     9181 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     9241 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     9301 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc
     9361 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg
     9421 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc
     9481 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
     9541 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
     9601 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
     9661 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
     9721 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
     9781 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc
     9841 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
     9901 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
     9961 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
    10021 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
    10081 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
    10141 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
    10201 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg
    10261 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga
    10321 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
    10381 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg
    10441 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg
    10501 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca
    10561 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccaga
//
