LOCUS       sequence-250501-       12145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of EGFR V292M.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12145)
  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 12145)
  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"; 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-EGFR-V292M
FEATURES             Location/Qualifiers
     source          1..12145
                     /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"
     CDS             join(3448..3519,3520..5382,5383..5451,5452..7080)
                     /codon_start=1
                     /product="precursor of human epidermal growth factor 
                     receptor"
                     /label=EGFR
                     /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE
                     DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL
                     QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ
                     WRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR
                     CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP
                     EGKYSFGATCMKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI
                     GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK
                     EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD
                     GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG
                     PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP
                     DNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG
                     CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP
                     SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(7082..7106)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             7159..7200
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    7221..7788
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    7238..7790
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7382..7399)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7609..7628
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(7802..7804,7805..7807,7808..8521)
                     /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             join(7802..7804,7805..7807,7808..8518)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7847..7868)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8108..8127)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8455..8476
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    8535..9123
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8588..8608)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9006..9017)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9198..9431
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9550..9568)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9636..9740
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9741..9809,9810..10601)
                     /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"
     rep_origin      10772..11360
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11261..11280
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11514..11531
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11771..11792
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11807..11812,11813..11830,11831..11837)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11845..11861
                     /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     11850..11872
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11869..11885
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11869..11885
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11935..11954
                     /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 cgaccctccg ggacggccgg ggcagcgctc
     3481 ctggcgctgc tggctgcgct ctgcccggcg agtcgggctc tggaggaaaa gaaagtttgc
     3541 caaggcacga gtaacaagct cacgcagttg ggcacttttg aagatcattt tctcagcctc
     3601 cagaggatgt tcaataactg tgaggtggtc cttgggaatt tggaaattac ctatgtgcag
     3661 aggaattatg atctttcctt cttaaagacc atccaggagg tggctggtta tgtcctcatt
     3721 gccctcaaca cagtggagcg aattcctttg gaaaacctgc agatcatcag aggaaatatg
     3781 tactacgaaa attcctatgc cttagcagtc ttatctaact atgatgcaaa taaaaccgga
     3841 ctgaaggagc tgcccatgag aaatttacag gaaatcctgc atggcgccgt gcggttcagc
     3901 aacaaccctg ccctgtgcaa cgtggagagc atccagtggc gggacatagt cagcagtgac
     3961 tttctcagca acatgtcgat ggacttccag aaccacctgg gcagctgcca aaagtgtgat
     4021 ccaagctgtc ccaatgggag ctgctggggt gcaggagagg agaactgcca gaaactgacc
     4081 aaaatcatct gtgcccagca gtgctccggg cgctgccgtg gcaagtcccc cagtgactgc
     4141 tgccacaacc agtgtgctgc aggctgcaca ggcccccggg agagcgactg cctggtctgc
     4201 cgcaaattcc gagacgaagc cacgtgcaag gacacctgcc ccccactcat gctctacaac
     4261 cccaccacgt accagatgga tgtgaacccc gagggcaaat acagctttgg tgccacctgc
     4321 atgaagaagt gtccccgtaa ttatgtggtg acagatcacg gctcgtgcgt ccgagcctgt
     4381 ggggccgaca gctatgagat ggaggaagac ggcgtccgca agtgtaagaa gtgcgaaggg
     4441 ccttgccgca aagtgtgtaa cggaataggt attggtgaat ttaaagactc actctccata
     4501 aatgctacga atattaaaca cttcaaaaac tgcacctcca tcagtggcga tctccacatc
     4561 ctgccggtgg catttagggg tgactccttc acacatactc ctcctctgga tccacaggaa
     4621 ctggatattc tgaaaaccgt aaaggaaatc acagggtttt tgctgattca ggcttggcct
     4681 gaaaacagga cggacctcca tgcctttgag aacctagaaa tcatacgcgg caggaccaag
     4741 caacatggtc agttttctct tgcagtcgtc agcctgaaca taacatcctt gggattacgc
     4801 tccctcaagg agataagtga tggagatgtg ataatttcag gaaacaaaaa tttgtgctat
     4861 gcaaatacaa taaactggaa aaaactgttt gggacctccg gtcagaaaac caaaattata
     4921 agcaacagag gtgaaaacag ctgcaaggcc acaggccagg tctgccatgc cttgtgctcc
     4981 cccgagggct gctggggccc ggagcccagg gactgcgtct cttgccggaa tgtcagccga
     5041 ggcagggaat gcgtggacaa gtgcaacctt ctggagggtg agccaaggga gtttgtggag
     5101 aactctgagt gcatacagtg ccacccagag tgcctgcctc aggccatgaa catcacctgc
     5161 acaggacggg gaccagacaa ctgtatccag tgtgcccact acattgacgg cccccactgc
     5221 gtcaagacct gcccggcagg agtcatggga gaaaacaaca ccctggtctg gaagtacgca
     5281 gacgccggcc atgtgtgcca cctgtgccat ccaaactgca cctacggatg cactgggcca
     5341 ggtcttgaag gctgtccaac gaatgggcct aagatcccgt ccatcgccac tgggatggtg
     5401 ggggccctcc tcttgctgct ggtggtggcc ctggggatcg gcctcttcat gcgaaggcgc
     5461 cacatcgttc ggaagcgcac gctgcggagg ctgctgcagg agagggagct tgtggagcct
     5521 cttacaccca gtggagaagc tcccaaccaa gctctcttga ggatcttgaa ggaaactgaa
     5581 ttcaaaaaga tcaaagtgct gggctccggt gcgttcggca cggtgtataa gggactctgg
     5641 atcccagaag gtgagaaagt taaaattccc gtcgctatca aggaattaag agaagcaaca
     5701 tctccgaaag ccaacaagga aatcctcgat gaagcctacg tgatggccag cgtggacaac
     5761 ccccacgtgt gccgcctgct gggcatctgc ctcacctcca ccgtgcagct catcacgcag
     5821 ctcatgccct tcggctgcct cctggactat gtccgggaac acaaagacaa tattggctcc
     5881 cagtacctgc tcaactggtg tgtgcagatc gcaaagggca tgaactactt ggaggaccgt
     5941 cgcttggtgc accgcgacct ggcagccagg aacgtactgg tgaaaacacc gcagcatgtc
     6001 aagatcacag attttgggct ggccaaactg ctgggtgcgg aagagaaaga ataccatgca
     6061 gaaggaggca aagtgcctat caagtggatg gcattggaat caattttaca cagaatctat
     6121 acccaccaga gtgatgtctg gagctacggg gtgaccgttt gggagttgat gacctttgga
     6181 tccaagccat atgacggaat ccctgccagc gagatctcct ccatcctgga gaaaggagaa
     6241 cgcctccctc agccacccat atgtaccatc gatgtctaca tgatcatggt caagtgctgg
     6301 atgatagacg cagatagtcg cccaaagttc cgtgagttga tcatcgaatt ctccaaaatg
     6361 gcccgagacc cccagcgcta ccttgtcatt cagggggatg aaagaatgca tttgccaagt
     6421 cctacagact ccaacttcta ccgtgccctg atggatgaag aagacatgga cgacgtggtg
     6481 gatgccgacg agtacctcat cccacagcag ggcttcttca gcagcccctc cacgtcacgg
     6541 actcccctcc tgagctctct gagtgcaacc agcaacaatt ccaccgtggc ttgcattgat
     6601 agaaatgggc tgcaaagctg tcccatcaag gaagacagct tcttgcagcg atacagctca
     6661 gaccccacag gcgccttgac tgaggacagc atagacgaca ccttcctccc agtgcctgaa
     6721 tacataaacc agtccgttcc caaaaggccc gctggctctg tgcagaatcc tgtctatcac
     6781 aatcagcctc tgaaccccgc gcccagcaga gacccacact accaggaccc ccacagcact
     6841 gcagtgggca accccgagta tctcaacact gtccagccca cctgtgtcaa cagcacattc
     6901 gacagccctg cccactgggc ccagaaaggc agccaccaaa ttagcctgga caaccctgac
     6961 taccagcagg acttctttcc caaggaagcc aagccaaatg gcatctttaa gggctccaca
     7021 gctgaaaatg cagaatacct aagggtcgcg ccacaaagca gtgaatttat tggagcatag
     7081 aacccagctt tcttgtacaa agtggttgat atccagcaca gtggcggccg ctcgagtcta
     7141 gagggcccgc ggttcgaagg taagcctatc cctaaccctc tcctcggtct cgattctacg
     7201 cgtaccggtt agtaatgaga tccctccccc ccccctaacg ttactggccg aagccgcttg
     7261 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc
     7321 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc
     7381 cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa
     7441 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct
     7501 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca
     7561 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca
     7621 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat
     7681 ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc
     7741 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaca
     7801 tatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     7861 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     7921 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     7981 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     8041 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     8101 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     8161 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     8221 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     8281 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     8341 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     8401 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     8461 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     8521 aatcgataga tcctaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     8581 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     8641 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     8701 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     8761 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     8821 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     8881 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     8941 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     9001 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     9061 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     9121 cgcctgagat cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     9181 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt
     9241 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     9301 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     9361 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     9421 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     9481 atgaatatca gagagtgaga ggcccgggtt aattaaggaa agggctagat cattcttgaa
     9541 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     9601 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     9661 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     9721 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     9781 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     9841 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     9901 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     9961 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
    10021 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
    10081 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
    10141 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
    10201 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
    10261 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
    10321 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
    10381 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
    10441 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
    10501 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
    10561 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
    10621 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
    10681 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
    10741 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
    10801 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
    10861 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
    10921 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
    10981 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
    11041 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
    11101 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
    11161 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
    11221 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
    11281 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
    11341 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
    11401 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
    11461 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
    11521 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
    11581 cgattcatta atgcagcaag ctcatggctg actaattttt tttatttatg cagaggccga
    11641 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
    11701 cttttgcaaa aagctccccg tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
    11761 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
    11821 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
    11881 atgacatgat tacgaatttc acaaataaag catttttttc actgcattct agttgtggtt
    11941 tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc
    12001 aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac ctgtggtttc
    12061 atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat tgtatttgtt
    12121 aaatatgtac tacaaactta gtagt
//
