LOCUS       sequence-250647-       12145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of EGFR P596L.
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-P596L
FEATURES             Location/Qualifiers
     source          1..12145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    217..334
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        382..1560
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          613..1551
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1508..1528
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1580..1604
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1607..1678,1679..3541,3542..3610,3611..5239)
                     /codon_start=1
                     /product="precursor of human epidermal growth factor 
                     receptor"
                     /label=EGFR
                     /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE
                     DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL
                     QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ
                     WRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR
                     CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP
                     EGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI
                     GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK
                     EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD
                     GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG
                     PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP
                     DNCIQCAHYIDGPHCVKTCLAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG
                     CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP
                     SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(5241..5265)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5318..5359
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    5380..5947
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5397..5949
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5541..5558)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5768..5787
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5961..5963,5964..5966,5967..6680)
                     /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(5961..5963,5964..5966,5967..6677)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6006..6027)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6267..6286)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6614..6635
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6694..7282
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6747..6767)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7165..7176)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7357..7590
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7709..7727)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7795..7899
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7900..7968,7969..8760)
                     /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      8931..9519
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9420..9439
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9673..9690
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9930..9951
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9966..9971,9972..9989,9990..9996)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10004..10020
                     /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     10009..10031
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10028..10044
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10028..10044
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10094..10113
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             10305..10938
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10985..11110
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11607..11840
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12025..12069
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
ORIGIN
        1 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
       61 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag
      121 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tctcgacggt
      181 atcgccgaat tcacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
      241 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
      301 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga
      361 tccagtttgg actagtcgtg aggctccggt gcccgtcagt gggcagagcg cacatcgccc
      421 acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg
      481 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
      541 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
      601 gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat
      661 ggcccttgcg tgccttgaat tacttccacc tggctgcagt acgtgattct tgatcccgag
      721 cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc cccttcgcct
      781 cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac
      841 cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt ttgatgacct
      901 gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga tctgcacact
      961 ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt
     1021 tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta gtctcaagct
     1081 ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca
     1141 aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct
     1201 gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca
     1261 caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg
     1321 gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc tttaggttgg
     1381 ggggaggggt tttatgcgat ggagtttccc cacactgagt gggtggagac tgaagttagg
     1441 ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg
     1501 ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttca ggtgtcgtga
     1561 agcggccctg cagatatcaa caagtttgta caaaaaagca ggcaccatgc gaccctccgg
     1621 gacggccggg gcagcgctcc tggcgctgct ggctgcgctc tgcccggcga gtcgggctct
     1681 ggaggaaaag aaagtttgcc aaggcacgag taacaagctc acgcagttgg gcacttttga
     1741 agatcatttt ctcagcctcc agaggatgtt caataactgt gaggtggtcc ttgggaattt
     1801 ggaaattacc tatgtgcaga ggaattatga tctttccttc ttaaagacca tccaggaggt
     1861 ggctggttat gtcctcattg ccctcaacac agtggagcga attcctttgg aaaacctgca
     1921 gatcatcaga ggaaatatgt actacgaaaa ttcctatgcc ttagcagtct tatctaacta
     1981 tgatgcaaat aaaaccggac tgaaggagct gcccatgaga aatttacagg aaatcctgca
     2041 tggcgccgtg cggttcagca acaaccctgc cctgtgcaac gtggagagca tccagtggcg
     2101 ggacatagtc agcagtgact ttctcagcaa catgtcgatg gacttccaga accacctggg
     2161 cagctgccaa aagtgtgatc caagctgtcc caatgggagc tgctggggtg caggagagga
     2221 gaactgccag aaactgacca aaatcatctg tgcccagcag tgctccgggc gctgccgtgg
     2281 caagtccccc agtgactgct gccacaacca gtgtgctgca ggctgcacag gcccccggga
     2341 gagcgactgc ctggtctgcc gcaaattccg agacgaagcc acgtgcaagg acacctgccc
     2401 cccactcatg ctctacaacc ccaccacgta ccagatggat gtgaaccccg agggcaaata
     2461 cagctttggt gccacctgcg tgaagaagtg tccccgtaat tatgtggtga cagatcacgg
     2521 ctcgtgcgtc cgagcctgtg gggccgacag ctatgagatg gaggaagacg gcgtccgcaa
     2581 gtgtaagaag tgcgaagggc cttgccgcaa agtgtgtaac ggaataggta ttggtgaatt
     2641 taaagactca ctctccataa atgctacgaa tattaaacac ttcaaaaact gcacctccat
     2701 cagtggcgat ctccacatcc tgccggtggc atttaggggt gactccttca cacatactcc
     2761 tcctctggat ccacaggaac tggatattct gaaaaccgta aaggaaatca cagggttttt
     2821 gctgattcag gcttggcctg aaaacaggac ggacctccat gcctttgaga acctagaaat
     2881 catacgcggc aggaccaagc aacatggtca gttttctctt gcagtcgtca gcctgaacat
     2941 aacatccttg ggattacgct ccctcaagga gataagtgat ggagatgtga taatttcagg
     3001 aaacaaaaat ttgtgctatg caaatacaat aaactggaaa aaactgtttg ggacctccgg
     3061 tcagaaaacc aaaattataa gcaacagagg tgaaaacagc tgcaaggcca caggccaggt
     3121 ctgccatgcc ttgtgctccc ccgagggctg ctggggcccg gagcccaggg actgcgtctc
     3181 ttgccggaat gtcagccgag gcagggaatg cgtggacaag tgcaaccttc tggagggtga
     3241 gccaagggag tttgtggaga actctgagtg catacagtgc cacccagagt gcctgcctca
     3301 ggccatgaac atcacctgca caggacgggg accagacaac tgtatccagt gtgcccacta
     3361 cattgacggc ccccactgcg tcaagacctg cctggcagga gtcatgggag aaaacaacac
     3421 cctggtctgg aagtacgcag acgccggcca tgtgtgccac ctgtgccatc caaactgcac
     3481 ctacggatgc actgggccag gtcttgaagg ctgtccaacg aatgggccta agatcccgtc
     3541 catcgccact gggatggtgg gggccctcct cttgctgctg gtggtggccc tggggatcgg
     3601 cctcttcatg cgaaggcgcc acatcgttcg gaagcgcacg ctgcggaggc tgctgcagga
     3661 gagggagctt gtggagcctc ttacacccag tggagaagct cccaaccaag ctctcttgag
     3721 gatcttgaag gaaactgaat tcaaaaagat caaagtgctg ggctccggtg cgttcggcac
     3781 ggtgtataag ggactctgga tcccagaagg tgagaaagtt aaaattcccg tcgctatcaa
     3841 ggaattaaga gaagcaacat ctccgaaagc caacaaggaa atcctcgatg aagcctacgt
     3901 gatggccagc gtggacaacc cccacgtgtg ccgcctgctg ggcatctgcc tcacctccac
     3961 cgtgcagctc atcacgcagc tcatgccctt cggctgcctc ctggactatg tccgggaaca
     4021 caaagacaat attggctccc agtacctgct caactggtgt gtgcagatcg caaagggcat
     4081 gaactacttg gaggaccgtc gcttggtgca ccgcgacctg gcagccagga acgtactggt
     4141 gaaaacaccg cagcatgtca agatcacaga ttttgggctg gccaaactgc tgggtgcgga
     4201 agagaaagaa taccatgcag aaggaggcaa agtgcctatc aagtggatgg cattggaatc
     4261 aattttacac agaatctata cccaccagag tgatgtctgg agctacgggg tgaccgtttg
     4321 ggagttgatg acctttggat ccaagccata tgacggaatc cctgccagcg agatctcctc
     4381 catcctggag aaaggagaac gcctccctca gccacccata tgtaccatcg atgtctacat
     4441 gatcatggtc aagtgctgga tgatagacgc agatagtcgc ccaaagttcc gtgagttgat
     4501 catcgaattc tccaaaatgg cccgagaccc ccagcgctac cttgtcattc agggggatga
     4561 aagaatgcat ttgccaagtc ctacagactc caacttctac cgtgccctga tggatgaaga
     4621 agacatggac gacgtggtgg atgccgacga gtacctcatc ccacagcagg gcttcttcag
     4681 cagcccctcc acgtcacgga ctcccctcct gagctctctg agtgcaacca gcaacaattc
     4741 caccgtggct tgcattgata gaaatgggct gcaaagctgt cccatcaagg aagacagctt
     4801 cttgcagcga tacagctcag accccacagg cgccttgact gaggacagca tagacgacac
     4861 cttcctccca gtgcctgaat acataaacca gtccgttccc aaaaggcccg ctggctctgt
     4921 gcagaatcct gtctatcaca atcagcctct gaaccccgcg cccagcagag acccacacta
     4981 ccaggacccc cacagcactg cagtgggcaa ccccgagtat ctcaacactg tccagcccac
     5041 ctgtgtcaac agcacattcg acagccctgc ccactgggcc cagaaaggca gccaccaaat
     5101 tagcctggac aaccctgact accagcagga cttctttccc aaggaagcca agccaaatgg
     5161 catctttaag ggctccacag ctgaaaatgc agaataccta agggtcgcgc cacaaagcag
     5221 tgaatttatt ggagcataga acccagcttt cttgtacaaa gtggttgata tccagcacag
     5281 tggcggccgc tcgagtctag agggcccgcg gttcgaaggt aagcctatcc ctaaccctct
     5341 cctcggtctc gattctacgc gtaccggtta gtaatgagat ccctcccccc cccctaacgt
     5401 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
     5461 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
     5521 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
     5581 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
     5641 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     5701 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     5761 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
     5821 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag
     5881 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     5941 atgataatat ggccacacat atggtgagca agggcgagga gctgttcacc ggggtggtgc
     6001 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     6061 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     6121 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     6181 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     6241 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     6301 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     6361 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     6421 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     6481 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     6541 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     6601 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     6661 tggacgagct gtacaagtaa atcgatagat cctaatcaac ctctggatta caaaatttgt
     6721 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     6781 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     6841 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     6901 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     6961 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     7021 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     7081 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     7141 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     7201 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     7261 tccctttggg ccgcctcccc gcctgagatc ctttaagacc aatgacttac aaggcagctg
     7321 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     7381 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     7441 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     7501 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     7561 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     7621 atttataact tgcaaagaaa tgaatatcag agagtgagag gcccgggtta attaaggaaa
     7681 gggctagatc attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     7741 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     7801 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     7861 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     7921 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     7981 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     8041 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     8101 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     8161 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     8221 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     8281 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     8341 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     8401 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     8461 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     8521 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     8581 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     8641 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     8701 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     8761 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     8821 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     8881 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     8941 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     9001 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     9061 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     9121 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     9181 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     9241 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     9301 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     9361 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     9421 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     9481 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     9541 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     9601 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     9661 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     9721 cctctccccg cgcgttggcc gattcattaa tgcagcaagc tcatggctga ctaatttttt
     9781 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     9841 gcttttttgg aggcctaggc ttttgcaaaa agctccccgt ggcacgacag gtttcccgac
     9901 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     9961 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
    10021 tttcacacag gaaacagcta tgacatgatt acgaatttca caaataaagc atttttttca
    10081 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaac
    10141 tggataactc aagctaacca aaatcatccc aaacttccca ccccataccc tattaccact
    10201 gccaattacc tgtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt
    10261 taaagaaatt gtatttgtta aatatgtact acaaacttag tagttggaag ggctaattca
    10321 ctcccaaaga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
    10381 tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct ttggatggtg
    10441 ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag gagagaacac
    10501 cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtgttaga
    10561 gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc atccggagta
    10621 cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg actttccagg
    10681 gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc
    10741 tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
    10801 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
    10861 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
    10921 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
    10981 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
    11041 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
    11101 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
    11161 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
    11221 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
    11281 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
    11341 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
    11401 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg
    11461 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
    11521 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
    11581 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
    11641 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
    11701 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
    11761 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
    11821 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
    11881 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
    11941 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
    12001 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
    12061 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcata
    12121 atgatagtag gaggcttggt aggtt
//
