LOCUS       Exported               11815 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Lentiviral expression of PIK3CA E542V.
ACCESSION   .
VERSION     .
KEYWORDS    pHAGE-PIK3CA-E542V
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11815)
  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 11815)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11815
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     CDS             6829..6870
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    6891..7458
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    6908..7460
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7052..7069)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7279..7298
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7472..8191
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             7472..8188
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7517..7538)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7778..7797)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8125..8146
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    8205..8793
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8258..8278)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8676..8687)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8868..9101
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9220..9238)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9306..9410
                     /gene="bla"
                     /label=AmpR promoter
     CDS             9411..10271
                     /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(9629..9648)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10442..11030
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10931..10950
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11184..11201
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     11293..11312
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     protein_bind    11441..11462
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        11477..11507
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11515..11531
                     /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     11520..11542
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11539..11555
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11539..11555
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11605..11624
                     /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 gtaatcactg 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 aaattattcg 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 taaaggatga aagttagagt
     6721 ttatgataag ccctatagtg atcgtattac acccaacttt tctaatacaa agtggttgat
     6781 atccagcaca gtggcggccg ctcgagtcta gagggcccgc ggttcgaagg taagcctatc
     6841 cctaaccctc tcctcggtct cgattctacg cgtaccggtt agtaatgaga tccctccccc
     6901 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     6961 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     7021 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
     7081 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     7141 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     7201 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     7261 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     7321 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt
     7381 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     7441 tgaaaaacac gatgataata tggccacaca tatggtgagc aagggcgagg agctgttcac
     7501 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     7561 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     7621 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     7681 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     7741 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     7801 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     7861 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     7921 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     7981 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     8041 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     8101 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     8161 cactctcggc atggacgagc tgtacaagta aatcgataga tcctaatcaa cctctggatt
     8221 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     8281 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     8341 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     8401 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     8461 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     8521 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     8581 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     8641 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     8701 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     8761 cgagtcggat ctccctttgg gccgcctccc cgcctgagat cctttaagac caatgactta
     8821 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     8881 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca
     8941 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     9001 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     9061 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
     9121 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggcccgggtt
     9181 aattaaggaa agggctagat cattcttgaa gacgaaaggg cctcgtgata cgcctatttt
     9241 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     9301 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     9361 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     9421 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     9481 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     9541 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     9601 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     9661 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     9721 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     9781 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     9841 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     9901 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     9961 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
    10021 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
    10081 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
    10141 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
    10201 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
    10261 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
    10321 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
    10381 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
    10441 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
    10501 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
    10561 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
    10621 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
    10681 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
    10741 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
    10801 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
    10861 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
    10921 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
    10981 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
    11041 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
    11101 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
    11161 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
    11221 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagcaag ctcatggctg
    11281 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
    11341 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctccccg tggcacgaca
    11401 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
    11461 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
    11521 gcggataaca atttcacaca ggaaacagct atgacatgat tacgaatttc acaaataaag
    11581 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
    11641 tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc
    11701 ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat tcctctgaat
    11761 tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt
//
