LOCUS       sequence-242895-       11696 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Lentiviral expression of PIK3CA R818C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11696)
  AUTHORS   Ng PK, Li J, Jeong KJ, Shao S, Chen H, Tsang YH, Sengupta S, Wang Z,
            Bhavana VH, Tran R, Soewito S, Minussi DC, Moreno D, Kong K, 
            Dogruluk T, Lu H, Gao J, Tokheim C, Zhou DC, Johnson AM, Zeng J, Ip 
            CKM, Ju Z, Wester M, Yu S, Li Y, Vellano CP, Schultz N, Karchin R, 
            Ding L, Lu Y, Cheung LWT, Chen K, Shaw KR, Meric-Bernstam F, Scott 
            KL, Yi S, Sahni N, Liang H, Mills GB
  TITLE     Systematic Functional Annotation of Somatic Mutations in Cancer.
  JOURNAL   Cancer Cell. 2018 Mar 12;33(3):450-462.e10. doi: 
            10.1016/j.ccell.2018.01.021.
  PUBMED    29533785
REFERENCE   2  (bases 1 to 11696)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.ccell.2018.01"; journalName: "Cancer Cell"; date: 
            "2018-03-12- 12"; volume: "33"; issue: "3"; pages: "450-462.e10"
COMMENT     Sequence Label: pHAGE-PIK3CA-R818C
FEATURES             Location/Qualifiers
     source          1..11696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2223..3401
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2454..3392
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3349..3369
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    3421..3445
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6656..6680)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        6688..6706
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6731..6750)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6732..6750)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(6756..6776)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             6830..6871
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    6892..7459
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    6909..7461
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7053..7070)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7280..7299
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7473..8072
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7473..7492)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7969..7989
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8086..8674
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8139..8159)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8557..8568)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8749..8982
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9101..9119)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9187..9291
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9292..9360,9361..10152)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(9510..9529)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10323..10911
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10812..10831
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11065..11082
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11322..11343
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11358..11363,11364..11381,11382..11388)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11396..11412
                     /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     11401..11423
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11420..11436
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11420..11436
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11486..11505
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcgt
     2221 gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     2281 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     2341 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     2401 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
     2461 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     2521 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
     2581 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
     2641 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
     2701 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
     2761 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
     2821 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
     2881 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
     2941 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     3001 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     3061 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     3121 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     3181 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     3241 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     3301 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     3361 cagtggttca aagttttttt cttccatttc aggtgtcgtg aagcggccct gcagatatca
     3421 acaagtttgt acaaaaaagc aggcaccatg cctccaagac catcatcagg tgaactgtgg
     3481 ggcatccact tgatgccccc aagaatccta gtagaatgtt tactaccaaa tggaatgata
     3541 gtgactttag aatgcctccg tgaggctaca ttaataacca taaagcatga actatttaaa
     3601 gaagcaagaa aataccccct ccatcaactt cttcaagatg aatcttctta cattttcgta
     3661 agtgttactc aagaagcaga aagggaagaa ttttttgatg aaacaagacg actttgtgac
     3721 cttcggcttt ttcaaccctt tttaaaagta attgaaccag taggcaaccg tgaagaaaag
     3781 atcctcaatc gagaaattgg ttttgctatc ggcatgccag tgtgtgaatt tgatatggtt
     3841 aaagatccag aagtacagga cttccgaaga aatattctga acgtttgtaa agaagctgtg
     3901 gatcttaggg acctcaattc acctcatagt agagcaatgt atgtctatcc tccaaatgta
     3961 gaatcttcac cagaattgcc aaagcacata tataataaat tagataaagg gcaaataata
     4021 gtggtgatct gggtaatagt ttctccaaat aatgacaagc agaagtatac tctgaaaatc
     4081 aaccatgact gtgtaccaga acaagtaatt gctgaagcaa tcaggaaaaa aactcgaagt
     4141 atgttgctat cctctgaaca actaaaactc tgtgttttag aatatcaggg caagtatatt
     4201 ttaaaagtgt gtggatgtga tgaatacttc ctagaaaaat atcctctgag tcagtataag
     4261 tatataagaa gctgtataat gcttgggagg atgcccaatt tgatgttgat ggctaaagaa
     4321 agcctttatt ctcaactgcc aatggactgt tttacaatgc catcttattc cagacgcatt
     4381 tccacagcta caccatatat gaatggagaa acatctacaa aatccctttg ggttataaat
     4441 agtgcactca gaataaaaat tctttgtgca acctacgtga atgtaaatat tcgagacatt
     4501 gataagatct atgttcgaac aggtatctac catggaggag aacccttatg tgacaatgtg
     4561 aacactcaaa gagtaccttg ttccaatccc aggtggaatg aatggctgaa ttatgatata
     4621 tacattcctg atcttcctcg tgctgctcga ctttgccttt ccatttgctc tgttaaaggc
     4681 cgaaagggtg ctaaagagga acactgtcca ttggcatggg gaaatataaa cttgtttgat
     4741 tacacagaca ctctagtatc tggaaaaatg gctttgaatc tttggccagt acctcatgga
     4801 ttagaagatt tgctgaaccc tattggtgtt actggatcaa atccaaataa agaaactcca
     4861 tgcttagagt tggagtttga ctggttcagc agtgtggtaa agttcccaga tatgtcagtg
     4921 attgaagagc atgccaattg gtctgtatcc cgagaagcag gatttagcta ttcccacgca
     4981 ggactgagta acagactagc tagagacaat gaattaaggg aaaatgacaa agaacagctc
     5041 aaagcaattt ctacacgaga tcctctctct gaaatcactg agcaggagaa agattttcta
     5101 tggagtcaca gacactattg tgtaactatc cccgaaattc tacccaaatt gcttctgtct
     5161 gttaaatgga attctagaga tgaagtagcc cagatgtatt gcttggtaaa agattggcct
     5221 ccaatcaaac ctgaacaggc tatggaactt ctggactgta attacccaga tcctatggtt
     5281 cgaggttttg ctgttcggtg cttggaaaaa tatttaacag atgacaaact ttctcagtat
     5341 ttaattcagc tagtacaggt cctaaaatat gaacaatatt tggataactt gcttgtgaga
     5401 tttttactga agaaagcatt gactaatcaa aggattgggc actttttctt ttggcattta
     5461 aaatctgaga tgcacaataa aacagttagc cagaggtttg gcctgctttt ggagtcctat
     5521 tgtcgtgcat gtgggatgta tttgaagcac ctgaataggc aagtcgaggc aatggaaaag
     5581 ctcattaact taactgacat tctcaaacag gagaagaagg atgaaacaca aaaggtacag
     5641 atgaagtttt tagttgagca aatgaggcga ccagatttca tggatgctct acagggcttt
     5701 ctgtctcctc taaaccctgc tcatcaacta ggaaacctca ggcttgaaga gtgtcgaatt
     5761 atgtcctctg caaaaaggcc actgtggttg aattgggaga acccagacat catgtcagag
     5821 ttactgtttc agaacaatga gatcatcttt aaaaatgggg atgatttacg gcaagatatg
     5881 ctaacacttc aaattatttg tattatggaa aatatctggc aaaatcaagg tcttgatctt
     5941 cgaatgttac cttatggttg tctgtcaatc ggtgactgtg tgggacttat tgaggtggtg
     6001 cgaaattctc acactattat gcaaattcag tgcaaaggcg gcttgaaagg tgcactgcag
     6061 ttcaacagcc acacactaca tcagtggctc aaagacaaga acaaaggaga aatatatgat
     6121 gcagccattg acctgtttac acgttcatgt gctggatact gtgtagctac cttcattttg
     6181 ggaattggag atcgtcacaa tagtaacatc atggtgaaag acgatggaca actgtttcat
     6241 atagattttg gacacttttt ggatcacaag aagaaaaaat ttggttataa acgagaacgt
     6301 gtgccatttg ttttgacaca ggatttctta atagtgatta gtaaaggagc ccaagaatgc
     6361 acaaagacaa gagaatttga gaggtttcag gagatgtgtt acaaggctta tctagctatt
     6421 cgacagcatg ccaatctctt cataaatctt ttctcaatga tgcttggctc tggaatgcca
     6481 gaactacaat cttttgatga cattgcatac attcgaaaga ccctagcctt agataaaact
     6541 gagcaagagg ctttggagta tttcatgaaa caaatgaatg atgcacatca tggtggctgg
     6601 acaacaaaaa tggattggat cttccacaca attaaacagc atgcattgaa ctagaaccca
     6661 gctttcttgt acaaagtggt gatatccaat taaccctcac taaaggtttg ttgttaagta
     6721 tgtgatttag ccctatagtg agtcgtatta ccacccaact tttctataca aagtggttga
     6781 tatccagcac agtggcggcc gctcgagtct agagggcccg cggttcgaag gtaagcctat
     6841 ccctaaccct ctcctcggtc tcgattctac gcgtaccggt tagtaatgag atccctcccc
     6901 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     6961 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     7021 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     7081 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc
     7141 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     7201 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     7261 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     7321 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
     7381 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     7441 ttgaaaaaca cgatgataat atggccacac atatgaccga gtacaagccc acggtgcgcc
     7501 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     7561 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     7621 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     7681 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     7741 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     7801 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     7861 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     7921 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     7981 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     8041 cctggtgcat gacccgcaag cccggtgcct aaatcgatag atcctaatca acctctggat
     8101 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     8161 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     8221 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     8281 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     8341 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     8401 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     8461 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     8521 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     8581 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     8641 acgagtcgga tctccctttg ggccgcctcc ccgcctgaga tcctttaaga ccaatgactt
     8701 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     8761 ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     8821 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     8881 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     8941 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     9001 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggcccgggt
     9061 taattaagga aagggctaga tcattcttga agacgaaagg gcctcgtgat acgcctattt
     9121 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     9181 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     9241 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     9301 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     9361 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     9421 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     9481 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     9541 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     9601 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     9661 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     9721 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     9781 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     9841 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     9901 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     9961 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
    10021 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
    10081 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
    10141 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
    10201 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
    10261 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
    10321 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
    10381 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
    10441 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
    10501 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
    10561 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
    10621 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
    10681 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
    10741 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
    10801 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
    10861 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
    10921 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
    10981 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
    11041 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
    11101 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagcaa gctcatggct
    11161 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
    11221 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccc gtggcacgac
    11281 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
    11341 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
    11401 agcggataac aatttcacac aggaaacagc tatgacatga ttacgaattt cacaaataaa
    11461 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
    11521 gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc caccccatac
    11581 cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa
    11641 ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagt
//
