LOCUS       sequence-443800-       10638 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This 3rd generation lentiviral plasmid expresses a S. pyogenes 
            CRISPR chimeric RNA element with customizable sgRNA from a U6 
            promoter and puromycin resistance from an EF-1a promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10638)
  AUTHORS   Stringer BW, Day BW, D'Souza RCJ, Jamieson PR, Ensbey KS, Bruce ZC, 
            Lim YC, Goasdoue K, Offenhauser C, Akgul S, Allan S, Robertson T, 
            Lucas P, Tollesson G, Campbell S, Winter C, Do H, Dobrovic A, Inglis
            PL, Jeffree RL, Johns TG, Boyd AW
  TITLE     A reference collection of patient-derived cell line and xenograft 
            models of proneural, classical and mesenchymal glioblastoma.
  JOURNAL   Sci Rep. 2019 Mar 20;9(1):4902. doi: 10.1038/s41598-019-41277-z.
  PUBMED    30894629
REFERENCE   2  (bases 1 to 10638)
  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: "Sci Rep."; 
            date: "2019-03-20"; volume: "9(1):4902. doi"; pages: " 
            10.1038/s41598-019-41277-z"
COMMENT     Sequence Label: lenti-sgRNA puro
FEATURES             Location/Qualifiers
     source          1..10638
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             51..284
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(310..327)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    316..540
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      586..1014
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(673..692)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     883..904
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1023..1043)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1028..1357
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1208..1343
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1270..1289
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1405..1452
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1471..1845
                     /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"
     polyA_signal    1975..2096
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2012..2031)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2066..2085
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2145..2161)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2145..2161)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2158..2180)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2169..2185
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2193..2199,2200..2217,2218..2223))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2238..2259
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2376..2393)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2547..3135)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2627..2646)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3306..4097,4098..4166))
                     /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     3929..3948
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4167..4271)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4346..4365)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4541..4920
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4922..5120
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5075..5095
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5138..5318
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5365..5490
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5983..6216
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6401..6445
                     /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    6743..6860
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6911..7159
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6911..6931
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7082..7101
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9041..9116
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        9178..9389
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             9414..10013
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPRVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     9910..9930
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10029..10617
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10082..10102)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10500..10511)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
       61 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
      121 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
      181 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
      241 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagggccc gtttaaaccc
      301 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
      361 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
      421 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
      481 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
      541 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
      601 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
      661 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
      721 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
      781 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
      841 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
      901 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
      961 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     1021 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     1081 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     1141 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     1201 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     1261 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     1321 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     1381 cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata
     1441 gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg
     1501 ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc
     1561 cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc
     1621 agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc
     1681 ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc
     1741 gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac
     1801 ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat
     1861 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     1921 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     1981 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2041 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     2101 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     2161 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     2221 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     2281 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     2341 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2401 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2461 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2521 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2581 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2641 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2701 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2761 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2821 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2881 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2941 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3001 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3061 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3121 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3181 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3241 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3301 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3361 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3421 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3481 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3541 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3601 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3661 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3721 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3781 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3841 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3901 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3961 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4021 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4081 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4141 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4201 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4261 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     4321 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     4381 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     4441 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
     4501 cgctgcttcg cgatgtacgg gccagatata cgcgcgcgtt gacattgatt attgactagt
     4561 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     4621 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     4681 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     4741 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     4801 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     4861 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     4921 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     4981 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     5041 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     5101 tgggaggtct atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat
     5161 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     5221 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     5281 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
     5341 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
     5401 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
     5461 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
     5521 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
     5581 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
     5641 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
     5701 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
     5761 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
     5821 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     5881 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     5941 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     6001 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     6061 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     6121 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     6181 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     6241 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     6301 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     6361 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     6421 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     6481 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     6541 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     6601 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     6661 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
     6721 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     6781 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     6841 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttaa
     6901 ttaaggtacc gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc
     6961 tgttagagag ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac
     7021 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
     7081 ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt
     7141 gtggaaagga cgaaacaccg gagacggttg taaatgagca cacaaaatac acatgctaaa
     7201 atattatatt ctatgacctt tataaaatca accaaaatct tctttttaat aactttagta
     7261 tcaataatta gaatttttat gttccttttt gcaaactttt aataaaaatg agcaaaataa
     7321 aaaaacgcta gttttagtaa ctcgcgttgt tttcttcacc tttaataata gctactccac
     7381 cacttgttcc taagcggtca gctcctgctt caatcatttt ttgagcatct tcaaatgttc
     7441 taactccacc agctgcttta actaaagcat tgtctttaac aactgacttc attagtttaa
     7501 catcttcaaa tgttgcacct gattttgaaa atcctgttga tgttttaaca aattctaatc
     7561 cagcttcaac agctatttca caagctttca tgatttcttc ttttgttaat aaacaatttt
     7621 ccataataca tttaacaaca tgtgatccag ctgctttttt tacagctttc atgtcttcta
     7681 aaactaattc ataatttttg tcttttaatg caccaatatt taataccata tcaatttctg
     7741 ttgcaccatc tttaattgct tcagaaactt cgaatgcttt tgtagctgtt gtgcatgcac
     7801 ctagaggaaa acctacaaca tttgttattc ctacatttgt gccttttaat aattctttac
     7861 aatagcttgt tcaatatgaa ttaacacaaa ctgttgcaaa atcaaattca attgcttcat
     7921 cacataattg tttaatttca gctttcgtag catcttgttt taataatgtg tgatctatat
     7981 atttgtttag tttcattttt tctcctatat attcattttt aattttaatt ctttaataat
     8041 ttcgtctact ttaactttag cgttttgaac agattcacca acacctataa aataaatttt
     8101 tagtttaggt tcagttccac ttgggcgaac agcaaatcat gacttatctt ctaaataaaa
     8161 ttttagtaag tcttgtcctg gcatattata cattccatcg atgtagtctt caacattaac
     8221 aactttaagt ccagcaattt gagttaaggg tgttgctctc aatgatttca ttaatggttc
     8281 aatttttaat ttcttttctt ctggtttaaa attcaagttt aaagtgaaag tgtaatatgc
     8341 acccatttct ttaaataaat cttctaaata gtctactaat gttttatttt gttttttata
     8401 aaatcaagca gcctctgcta ttaatataga agcttgtatt ccatctttat ctctagctga
     8461 gtcatcaatt acatatccat aactttcttc ataagcaaaa acaaaattta atccgttatc
     8521 ttcttcttta gcaatttctc tacccattca tttaaatcca gttaaagttt ttacaatatt
     8581 aactccatat ttttcatgag cgattctatc acccaaatca cttgttacaa aacttgaata
     8641 tagagccgga ttttttggaa tgctatttaa gcgttttaga tttgataatt ttcaatcaat
     8701 taaaattggt cctgtttgat ttccatctaa tcttacaaaa tgaccatcat gttttattgc
     8761 cattccaaat ctgtcagcat ctgggtcatt cataataata atatctgcat catgtttaat
     8821 accatattca agcggtattt ttcatgcagg atcaaattct ggatttggat ttacaacatt
     8881 tttaaatgtt tcatcttcaa atgcatgctc ttcaacctca ataacgttat atcctgattc
     8941 acgtaatatt tttggggtaa atttagttcc tgttccatta actgcgctaa aaataatttt
     9001 taaatctttt ttagcttctt gctctttttt gtacgtctct gttttagagc tagaaatagc
     9061 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     9121 ttgaattcgc tagctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg
     9181 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg
     9241 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     9301 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     9361 ttcgcaacgg gtttgccgcc agaacacagg accggttcta gagcgctgcc accatgaccg
     9421 agtacaagcc cagggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     9481 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     9541 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     9601 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     9661 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     9721 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     9781 ggttcctggc caccgtcgga gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     9841 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     9901 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     9961 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgaacgcgtt
    10021 aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
    10081 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
    10141 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
    10201 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
    10261 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
    10321 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
    10381 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
    10441 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
    10501 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
    10561 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcgtc
    10621 gactttaaga ccaatgac
//
