LOCUS       sequence-250547-       12122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of EGFR G719C.
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-G719C
FEATURES             Location/Qualifiers
     source          1..12122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    239..356
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        404..1582
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          635..1573
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1530..1550
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1602..1626
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1629..1700,1701..3563,3564..3632,3633..5261)
                     /codon_start=1
                     /product="precursor of human epidermal growth factor 
                     receptor"
                     /label=EGFR
                     /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE
                     DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL
                     QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ
                     WRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR
                     CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP
                     EGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI
                     GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK
                     EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD
                     GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG
                     PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP
                     DNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG
                     CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP
                     SGEAPNQALLRILKETEFKKIKVLCSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(5263..5287)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        5295..5313
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5338..5357)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5339..5357)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(5363..5383)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             5437..5478
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    5499..6066
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5516..6068
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5660..5677)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5887..5906
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6080..6679
                     /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(6080..6099)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6576..6596
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6693..7281
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6746..6766)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7164..7175)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7356..7589
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7708..7726)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7794..7898
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7899..7967,7968..8759)
                     /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      8930..9518
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9419..9438
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9672..9689
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9929..9950
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9965..9970,9971..9988,9989..9995)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10003..10019
                     /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     10008..10030
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10027..10043
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10027..10043
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10093..10112
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             10304..10937
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10984..11109
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11606..11839
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12024..12068
                     /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 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga
       61 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc
      121 gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag atccattcga
      181 ttagtgaacg gatctcgacg gtatcgccga attcacaaat ggcagtattc atccacaatt
      241 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
      301 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg
      361 tttattacag ggacagcaga gatccagttt ggactagtcg tgaggctccg gtgcccgtca
      421 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
      481 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
      541 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
      601 tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg
      661 gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca cctggctgca
      721 gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg
      781 cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg
      841 ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc
      901 catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa
      961 tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc
     1021 ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat
     1081 cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg
     1141 tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag
     1201 atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga
     1261 gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc
     1321 atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg
     1381 gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga
     1441 gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc
     1501 cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt
     1561 tcttccattt caggtgtcgt gaagcggccc tgcagatatc aacaagtttg tacaaaaaag
     1621 caggcaccat gcgaccctcc gggacggccg gggcagcgct cctggcgctg ctggctgcgc
     1681 tctgcccggc gagtcgggct ctggaggaaa agaaagtttg ccaaggcacg agtaacaagc
     1741 tcacgcagtt gggcactttt gaagatcatt ttctcagcct ccagaggatg ttcaataact
     1801 gtgaggtggt ccttgggaat ttggaaatta cctatgtgca gaggaattat gatctttcct
     1861 tcttaaagac catccaggag gtggctggtt atgtcctcat tgccctcaac acagtggagc
     1921 gaattccttt ggaaaacctg cagatcatca gaggaaatat gtactacgaa aattcctatg
     1981 ccttagcagt cttatctaac tatgatgcaa ataaaaccgg actgaaggag ctgcccatga
     2041 gaaatttaca ggaaatcctg catggcgccg tgcggttcag caacaaccct gccctgtgca
     2101 acgtggagag catccagtgg cgggacatag tcagcagtga ctttctcagc aacatgtcga
     2161 tggacttcca gaaccacctg ggcagctgcc aaaagtgtga tccaagctgt cccaatggga
     2221 gctgctgggg tgcaggagag gagaactgcc agaaactgac caaaatcatc tgtgcccagc
     2281 agtgctccgg gcgctgccgt ggcaagtccc ccagtgactg ctgccacaac cagtgtgctg
     2341 caggctgcac aggcccccgg gagagcgact gcctggtctg ccgcaaattc cgagacgaag
     2401 ccacgtgcaa ggacacctgc cccccactca tgctctacaa ccccaccacg taccagatgg
     2461 atgtgaaccc cgagggcaaa tacagctttg gtgccacctg cgtgaagaag tgtccccgta
     2521 attatgtggt gacagatcac ggctcgtgcg tccgagcctg tggggccgac agctatgaga
     2581 tggaggaaga cggcgtccgc aagtgtaaga agtgcgaagg gccttgccgc aaagtgtgta
     2641 acggaatagg tattggtgaa tttaaagact cactctccat aaatgctacg aatattaaac
     2701 acttcaaaaa ctgcacctcc atcagtggcg atctccacat cctgccggtg gcatttaggg
     2761 gtgactcctt cacacatact cctcctctgg atccacagga actggatatt ctgaaaaccg
     2821 taaaggaaat cacagggttt ttgctgattc aggcttggcc tgaaaacagg acggacctcc
     2881 atgcctttga gaacctagaa atcatacgcg gcaggaccaa gcaacatggt cagttttctc
     2941 ttgcagtcgt cagcctgaac ataacatcct tgggattacg ctccctcaag gagataagtg
     3001 atggagatgt gataatttca ggaaacaaaa atttgtgcta tgcaaataca ataaactgga
     3061 aaaaactgtt tgggacctcc ggtcagaaaa ccaaaattat aagcaacaga ggtgaaaaca
     3121 gctgcaaggc cacaggccag gtctgccatg ccttgtgctc ccccgagggc tgctggggcc
     3181 cggagcccag ggactgcgtc tcttgccgga atgtcagccg aggcagggaa tgcgtggaca
     3241 agtgcaacct tctggagggt gagccaaggg agtttgtgga gaactctgag tgcatacagt
     3301 gccacccaga gtgcctgcct caggccatga acatcacctg cacaggacgg ggaccagaca
     3361 actgtatcca gtgtgcccac tacattgacg gcccccactg cgtcaagacc tgcccggcag
     3421 gagtcatggg agaaaacaac accctggtct ggaagtacgc agacgccggc catgtgtgcc
     3481 acctgtgcca tccaaactgc acctacggat gcactgggcc aggtcttgaa ggctgtccaa
     3541 cgaatgggcc taagatcccg tccatcgcca ctgggatggt gggggccctc ctcttgctgc
     3601 tggtggtggc cctggggatc ggcctcttca tgcgaaggcg ccacatcgtt cggaagcgca
     3661 cgctgcggag gctgctgcag gagagggagc ttgtggagcc tcttacaccc agtggagaag
     3721 ctcccaacca agctctcttg aggatcttga aggaaactga attcaaaaag atcaaagtgc
     3781 tgtgctccgg tgcgttcggc acggtgtata agggactctg gatcccagaa ggtgagaaag
     3841 ttaaaattcc cgtcgctatc aaggaattaa gagaagcaac atctccgaaa gccaacaagg
     3901 aaatcctcga tgaagcctac gtgatggcca gcgtggacaa cccccacgtg tgccgcctgc
     3961 tgggcatctg cctcacctcc accgtgcagc tcatcacgca gctcatgccc ttcggctgcc
     4021 tcctggacta tgtccgggaa cacaaagaca atattggctc ccagtacctg ctcaactggt
     4081 gtgtgcagat cgcaaagggc atgaactact tggaggaccg tcgcttggtg caccgcgacc
     4141 tggcagccag gaacgtactg gtgaaaacac cgcagcatgt caagatcaca gattttgggc
     4201 tggccaaact gctgggtgcg gaagagaaag aataccatgc agaaggaggc aaagtgccta
     4261 tcaagtggat ggcattggaa tcaattttac acagaatcta tacccaccag agtgatgtct
     4321 ggagctacgg ggtgaccgtt tgggagttga tgacctttgg atccaagcca tatgacggaa
     4381 tccctgccag cgagatctcc tccatcctgg agaaaggaga acgcctccct cagccaccca
     4441 tatgtaccat cgatgtctac atgatcatgg tcaagtgctg gatgatagac gcagatagtc
     4501 gcccaaagtt ccgtgagttg atcatcgaat tctccaaaat ggcccgagac ccccagcgct
     4561 accttgtcat tcagggggat gaaagaatgc atttgccaag tcctacagac tccaacttct
     4621 accgtgccct gatggatgaa gaagacatgg acgacgtggt ggatgccgac gagtacctca
     4681 tcccacagca gggcttcttc agcagcccct ccacgtcacg gactcccctc ctgagctctc
     4741 tgagtgcaac cagcaacaat tccaccgtgg cttgcattga tagaaatggg ctgcaaagct
     4801 gtcccatcaa ggaagacagc ttcttgcagc gatacagctc agaccccaca ggcgccttga
     4861 ctgaggacag catagacgac accttcctcc cagtgcctga atacataaac cagtccgttc
     4921 ccaaaaggcc cgctggctct gtgcagaatc ctgtctatca caatcagcct ctgaaccccg
     4981 cgcccagcag agacccacac taccaggacc cccacagcac tgcagtgggc aaccccgagt
     5041 atctcaacac tgtccagccc acctgtgtca acagcacatt cgacagccct gcccactggg
     5101 cccagaaagg cagccaccaa attagcctgg acaaccctga ctaccagcag gacttctttc
     5161 ccaaggaagc caagccaaat ggcatcttta agggctccac agctgaaaat gcagaatacc
     5221 taagggtcgc gccacaaagc agtgaattta ttggagcata gaacccagct ttcttgtaca
     5281 aagtggtgat atccaattaa ccctcactaa aggtgaaatg tgtatttgta tgtttagccc
     5341 tatagtgagt cgtattacca cccaactttt ctatacaaag tggttgatat ccagcacagt
     5401 ggcggccgct cgagtctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc
     5461 ctcggtctcg attctacgcg taccggttag taatgagatc cctccccccc ccctaacgtt
     5521 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     5581 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     5641 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     5701 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     5761 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     5821 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     5881 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     5941 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg
     6001 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     6061 tgataatatg gccacacata tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     6121 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     6181 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     6241 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     6301 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     6361 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     6421 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     6481 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     6541 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     6601 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     6661 ccgcaagccc ggtgcctaaa tcgatagatc ctaatcaacc tctggattac aaaatttgtg
     6721 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     6781 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     6841 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     6901 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     6961 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     7021 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     7081 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     7141 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     7201 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     7261 ccctttgggc cgcctccccg cctgagatcc tttaagacca atgacttaca aggcagctgt
     7321 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     7381 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     7441 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     7501 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     7561 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     7621 tttataactt gcaaagaaat gaatatcaga gagtgagagg cccgggttaa ttaaggaaag
     7681 ggctagatca ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     7741 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     7801 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     7861 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     7921 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     7981 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     8041 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     8101 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     8161 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     8221 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     8281 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     8341 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     8401 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     8461 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     8521 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     8581 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     8641 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     8701 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     8761 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     8821 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     8881 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     8941 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     9001 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     9061 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     9121 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     9181 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     9241 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     9301 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     9361 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     9421 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     9481 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     9541 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     9601 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     9661 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     9721 ctctccccgc gcgttggccg attcattaat gcagcaagct catggctgac taattttttt
     9781 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     9841 cttttttgga ggcctaggct tttgcaaaaa gctccccgtg gcacgacagg tttcccgact
     9901 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     9961 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
    10021 ttcacacagg aaacagctat gacatgatta cgaatttcac aaataaagca tttttttcac
    10081 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaact
    10141 ggataactca agctaaccaa aatcatccca aacttcccac cccataccct attaccactg
    10201 ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta ttttcatttt
    10261 aaagaaattg tatttgttaa atatgtacta caaacttagt agttggaagg gctaattcac
    10321 tcccaaagaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct
    10381 gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc
    10441 tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc
    10501 agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag
    10561 tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac
    10621 ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg
    10681 aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct
    10741 gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
    10801 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
    10861 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
    10921 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag
    10981 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
    11041 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
    11101 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg
    11161 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa
    11221 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga
    11281 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta
    11341 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac
    11401 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cggccgctga
    11461 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
    11521 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
    11581 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
    11641 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
    11701 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
    11761 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
    11821 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
    11881 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
    11941 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
    12001 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
    12061 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
    12121 tg
//
