LOCUS       sequence-250629-       12122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of EGFR N444S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12122)
  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 12122)
  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-N444S
FEATURES             Location/Qualifiers
     source          1..12122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    189..306
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        354..1532
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          585..1523
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1480..1500
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1552..1576
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1579..1650,1651..3513,3514..3582,3583..5211)
                     /codon_start=1
                     /product="precursor of human epidermal growth factor 
                     receptor"
                     /label=EGFR
                     /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE
                     DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL
                     QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ
                     WRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR
                     CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP
                     EGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI
                     GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK
                     EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLSITSLGLRSLKEISD
                     GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG
                     PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP
                     DNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG
                     CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP
                     SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(5213..5237)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        5245..5263
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5288..5307)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5289..5307)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(5313..5333)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             5387..5428
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    5449..6016
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5466..6018
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5610..5627)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5837..5856
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6030..6629
                     /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(6030..6049)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6526..6546
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6643..7231
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6696..6716)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7114..7125)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7306..7539
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7658..7676)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7744..7848
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7849..7917,7918..8709)
                     /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      8880..9468
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9369..9388
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9622..9639
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9879..9900
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9915..9920,9921..9938,9939..9945)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9953..9969
                     /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     9958..9980
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9977..9993
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9977..9993
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10043..10062
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             10254..10887
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10934..11059
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11556..11789
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11974..12018
                     /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 agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc
       61 gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag
      121 atccattcga ttagtgaacg gatctcgacg gtatcgccga attcacaaat ggcagtattc
      181 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
      241 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
      301 aattttcggg tttattacag ggacagcaga gatccagttt ggactagtcg tgaggctccg
      361 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
      421 tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
      481 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
      541 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg
      601 gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca
      661 cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt
      721 cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc
      781 gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata
      841 agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata
      901 gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg
      961 gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc
     1021 caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg
     1081 cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt
     1141 gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc
     1201 gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag
     1261 ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct
     1321 cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc
     1381 cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct
     1441 tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc
     1501 aaagtttttt tcttccattt caggtgtcgt gaagcggccc tgcagatatc aacaagtttg
     1561 tacaaaaaag caggcaccat gcgaccctcc gggacggccg gggcagcgct cctggcgctg
     1621 ctggctgcgc tctgcccggc gagtcgggct ctggaggaaa agaaagtttg ccaaggcacg
     1681 agtaacaagc tcacgcagtt gggcactttt gaagatcatt ttctcagcct ccagaggatg
     1741 ttcaataact gtgaggtggt ccttgggaat ttggaaatta cctatgtgca gaggaattat
     1801 gatctttcct tcttaaagac catccaggag gtggctggtt atgtcctcat tgccctcaac
     1861 acagtggagc gaattccttt ggaaaacctg cagatcatca gaggaaatat gtactacgaa
     1921 aattcctatg ccttagcagt cttatctaac tatgatgcaa ataaaaccgg actgaaggag
     1981 ctgcccatga gaaatttaca ggaaatcctg catggcgccg tgcggttcag caacaaccct
     2041 gccctgtgca acgtggagag catccagtgg cgggacatag tcagcagtga ctttctcagc
     2101 aacatgtcga tggacttcca gaaccacctg ggcagctgcc aaaagtgtga tccaagctgt
     2161 cccaatggga gctgctgggg tgcaggagag gagaactgcc agaaactgac caaaatcatc
     2221 tgtgcccagc agtgctccgg gcgctgccgt ggcaagtccc ccagtgactg ctgccacaac
     2281 cagtgtgctg caggctgcac aggcccccgg gagagcgact gcctggtctg ccgcaaattc
     2341 cgagacgaag ccacgtgcaa ggacacctgc cccccactca tgctctacaa ccccaccacg
     2401 taccagatgg atgtgaaccc cgagggcaaa tacagctttg gtgccacctg cgtgaagaag
     2461 tgtccccgta attatgtggt gacagatcac ggctcgtgcg tccgagcctg tggggccgac
     2521 agctatgaga tggaggaaga cggcgtccgc aagtgtaaga agtgcgaagg gccttgccgc
     2581 aaagtgtgta acggaatagg tattggtgaa tttaaagact cactctccat aaatgctacg
     2641 aatattaaac acttcaaaaa ctgcacctcc atcagtggcg atctccacat cctgccggtg
     2701 gcatttaggg gtgactcctt cacacatact cctcctctgg atccacagga actggatatt
     2761 ctgaaaaccg taaaggaaat cacagggttt ttgctgattc aggcttggcc tgaaaacagg
     2821 acggacctcc atgcctttga gaacctagaa atcatacgcg gcaggaccaa gcaacatggt
     2881 cagttttctc ttgcagtcgt cagcctgagc ataacatcct tgggattacg ctccctcaag
     2941 gagataagtg atggagatgt gataatttca ggaaacaaaa atttgtgcta tgcaaataca
     3001 ataaactgga aaaaactgtt tgggacctcc ggtcagaaaa ccaaaattat aagcaacaga
     3061 ggtgaaaaca gctgcaaggc cacaggccag gtctgccatg ccttgtgctc ccccgagggc
     3121 tgctggggcc cggagcccag ggactgcgtc tcttgccgga atgtcagccg aggcagggaa
     3181 tgcgtggaca agtgcaacct tctggagggt gagccaaggg agtttgtgga gaactctgag
     3241 tgcatacagt gccacccaga gtgcctgcct caggccatga acatcacctg cacaggacgg
     3301 ggaccagaca actgtatcca gtgtgcccac tacattgacg gcccccactg cgtcaagacc
     3361 tgcccggcag gagtcatggg agaaaacaac accctggtct ggaagtacgc agacgccggc
     3421 catgtgtgcc acctgtgcca tccaaactgc acctacggat gcactgggcc aggtcttgaa
     3481 ggctgtccaa cgaatgggcc taagatcccg tccatcgcca ctgggatggt gggggccctc
     3541 ctcttgctgc tggtggtggc cctggggatc ggcctcttca tgcgaaggcg ccacatcgtt
     3601 cggaagcgca cgctgcggag gctgctgcag gagagggagc ttgtggagcc tcttacaccc
     3661 agtggagaag ctcccaacca agctctcttg aggatcttga aggaaactga attcaaaaag
     3721 atcaaagtgc tgggctccgg tgcgttcggc acggtgtata agggactctg gatcccagaa
     3781 ggtgagaaag ttaaaattcc cgtcgctatc aaggaattaa gagaagcaac atctccgaaa
     3841 gccaacaagg aaatcctcga tgaagcctac gtgatggcca gcgtggacaa cccccacgtg
     3901 tgccgcctgc tgggcatctg cctcacctcc accgtgcagc tcatcacgca gctaatgccc
     3961 ttcggctgcc tcctggacta tgtccgggaa cacaaagaca atattggctc ccagtacctg
     4021 ctcaactggt gtgtgcagat cgcaaagggc atgaactact tggaggaccg tcgcttggtg
     4081 caccgcgacc tggcagccag gaacgtactg gtgaaaacac cgcagcatgt caagatcaca
     4141 gattttgggc tggccaaact gctgggtgcg gaagagaaag aataccatgc agaaggaggc
     4201 aaagtgccta tcaagtggat ggcattggaa tcaattttac acagaatcta tacccaccag
     4261 agtgatgtct ggagctacgg ggtgaccgtt tgggagttga tgacctttgg atccaagcca
     4321 tatgacggaa tccctgccag cgagatctcc tccatcctgg agaaaggaga acgcctccct
     4381 cagccaccca tatgtaccat cgatgtctac atgatcatgg tcaagtgctg gatgatagac
     4441 gcagatagtc gcccaaagtt ccgtgagttg atcatcgaat tctccaaaat ggcccgagac
     4501 ccccagcgct accttgtcat tcagggggat gaaagaatgc atttgccaag tcctacagac
     4561 tccaacttct accgtgccct gatggatgaa gaagacatgg acgacgtggt ggatgccgac
     4621 gagtacctca tcccacagca gggcttcttc agcagcccct ccacgtcacg gactcccctc
     4681 ctgagctctc tgagtgcaac cagcaacaat tccaccgtgg cttgcattga tagaaatggg
     4741 ctgcaaagct gtcccatcaa ggaagacagc ttcttgcagc gatacagctc agaccccaca
     4801 ggcgccttga ctgaggacag catagacgac accttcctcc cagtgcctga atacataaac
     4861 cagtccgttc ccaaaaggcc cgctggctct gtgcagaatc ctgtctatca caatcagcct
     4921 ctgaaccccg cgcccagcag agacccacac taccaggacc cccacagcac tgcagtgggc
     4981 aaccccgagt atctcaacac tgtccagccc acctgtgtca acagcacatt cgacagccct
     5041 gcccactggg cccagaaagg cagccaccaa attagcctgg acaaccctga ctaccagcag
     5101 gacttctttc ccaaggaagc caagccaaat ggcatcttta agggctccac agctgaaaat
     5161 gcagaatacc taagggtcgc gccacaaagc agtgaattta ttggagcata gaacccagct
     5221 ttcttgtaca aagtggtgat atccaattaa ccctcactaa aggtagttaa agtatagttg
     5281 tttgagtccc tatagtgagt cgtattacca cccaactttt ctatacaaag tggttgatat
     5341 ccagcacagt ggcggccgct cgagtctaga gggcccgcgg ttcgaaggta agcctatccc
     5401 taaccctctc ctcggtctcg attctacgcg taccggttag taatgagatc cctccccccc
     5461 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     5521 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     5581 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     5641 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     5701 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     5761 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     5821 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     5881 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     5941 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     6001 aaaaacacga tgataatatg gccacacata tgaccgagta caagcccacg gtgcgcctcg
     6061 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     6121 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     6181 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     6241 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     6301 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     6361 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     6421 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     6481 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     6541 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     6601 ggtgcatgac ccgcaagccc ggtgcctaaa tcgatagatc ctaatcaacc tctggattac
     6661 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     6721 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     6781 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     6841 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     6901 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     6961 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     7021 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     7081 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     7141 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     7201 agtcggatct ccctttgggc cgcctccccg cctgagatcc tttaagacca atgacttaca
     7261 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     7321 actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga
     7381 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     7441 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     7501 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta
     7561 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg cccgggttaa
     7621 ttaaggaaag ggctagatca ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     7681 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     7741 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     7801 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     7861 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     7921 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     7981 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     8041 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     8101 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     8161 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     8221 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     8281 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     8341 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     8401 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     8461 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     8521 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     8581 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     8641 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     8701 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     8761 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     8821 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     8881 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     8941 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     9001 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     9061 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     9121 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     9181 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     9241 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     9301 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     9361 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     9421 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     9481 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     9541 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     9601 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     9661 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagcaagct catggctgac
     9721 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     9781 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctccccgtg gcacgacagg
     9841 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     9901 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     9961 ggataacaat ttcacacagg aaacagctat gacatgatta cgaatttcac aaataaagca
    10021 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
    10081 tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac cccataccct
    10141 attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta
    10201 ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt agttggaagg
    10261 gctaattcac tcccaaagaa gacaagatat ccttgatctg tggatctacc acacacaagg
    10321 ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt
    10381 tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg
    10441 agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
    10501 agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca
    10561 tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga
    10621 ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat
    10681 ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg
    10741 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
    10801 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
    10861 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
    10921 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
    10981 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
    11041 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
    11101 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
    11161 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
    11221 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
    11281 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
    11341 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
    11401 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
    11461 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
    11521 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
    11581 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
    11641 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
    11701 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
    11761 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
    11821 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
    11881 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
    11941 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
    12001 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
    12061 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
    12121 at
//
