LentiGuide-zeo

LOCUS       LentiGuide-zeo               9964 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses S. pyogenes CRISPR chimeric RNA element with customizable 
            sgRNA from U6 promoter and zeocin resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    LentiGuide-zeo
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9964)
  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 9964)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..9964
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..241
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     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..4089
                     /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"
     misc_feature    4105..4693
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4158..4178)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4576..4587)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4765..4998
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5076..5197
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5113..5132)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5167..5186
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5237..5372
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5299..5318
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(5392..5411)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5393..5411)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5421..5438)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5421..5437)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5430..5452)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5579..6034
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5666..5685)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5876..5897
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6060..6164
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6165..7025
                     /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(6383..6402)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7196..7784
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7685..7704
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7938..7955
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8072..8093
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8108..8138
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8146..8162
                     /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     8151..8173
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8170..8186
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8170..8186
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8205..8225
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8207..8225
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8253..8479
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8480..8660
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8707..8832
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9325..9558
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9743..9787
                     /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 ggtcccgtgc 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 cgaaatggcc
     3721 aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc
     3781 tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc
     3841 cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc
     3901 ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg
     3961 tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg
     4021 gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag
     4081 caggactgaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     4141 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     4201 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     4261 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     4321 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     4381 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     4441 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     4501 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     4561 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     4621 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     4681 gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     4741 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     4801 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     4861 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     4921 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     4981 tgtggaaaat ctctagcagt acgtatagta gttcatgtca tcttattatt cagtatttat
     5041 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     5101 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5161 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     5221 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5281 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     5341 ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt
     5401 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     5461 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     5521 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     5581 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5641 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5701 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5761 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5821 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5881 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5941 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     6001 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     6061 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     6121 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     6181 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     6241 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     6301 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     6361 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     6421 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     6481 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     6541 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     6601 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     6661 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     6721 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     6781 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     6841 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6901 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6961 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     7021 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     7081 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7141 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7201 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7261 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7321 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     7381 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7441 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     7501 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     7561 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     7621 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     7681 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     7741 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7801 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     7861 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     7921 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     7981 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     8041 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     8101 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     8161 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     8221 aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc
     8281 ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc
     8341 atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg
     8401 tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg
     8461 cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc
     8521 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     8581 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     8641 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
     8701 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
     8761 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
     8821 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
     8881 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     8941 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     9001 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     9061 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
     9121 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat
     9181 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     9241 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     9301 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     9361 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     9421 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     9481 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     9541 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     9601 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     9661 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     9721 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     9781 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     9841 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     9901 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     9961 caga
//