LOCUS       sequence-250690-       12122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of EGFR R222C.
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-R222C
FEATURES             Location/Qualifiers
     source          1..12122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    152..269
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        317..1495
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          548..1486
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1443..1463
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1515..1539
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1542..1613,1614..3476,3477..3545,3546..5174)
                     /codon_start=1
                     /product="precursor of human epidermal growth factor 
                     receptor"
                     /label=EGFR
                     /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE
                     DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL
                     QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ
                     WRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGC
                     CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP
                     EGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI
                     GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK
                     EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD
                     GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG
                     PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP
                     DNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG
                     CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP
                     SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(5176..5200)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        5208..5226
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5251..5270)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5252..5270)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(5276..5296)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             5350..5391
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    5412..5979
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5429..5981
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5573..5590)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5800..5819
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5993..6592
                     /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(5993..6012)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6489..6509
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6606..7194
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6659..6679)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7077..7088)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7269..7502
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7621..7639)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7707..7811
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7812..7880,7881..8672)
                     /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      8843..9431
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9332..9351
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9585..9602
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9842..9863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9878..9883,9884..9901,9902..9908)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9916..9932
                     /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     9921..9943
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9940..9956
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9940..9956
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10006..10025
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             10217..10850
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10897..11022
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11519..11752
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11937..11981
                     /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 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
       61 ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg acggtatcgc
      121 cgaattcaca aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt
      181 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
      241 aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag
      301 tttggactag tcgtgaggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
      361 ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg
      421 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
      481 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
      541 aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc
      601 ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc ccgagcttcg
      661 ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc
      721 ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg
      781 cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc
      841 gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat
      901 ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc
      961 gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg
     1021 gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct
     1081 ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg
     1141 gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag
     1201 gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc
     1261 gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga
     1321 ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc
     1381 ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat
     1441 tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaagcgg
     1501 ccctgcagat atcaacaagt ttgtacaaaa aagcaggcac catgcgaccc tccgggacgg
     1561 ccggggcagc gctcctggcg ctgctggctg cgctctgccc ggcgagtcgg gctctggagg
     1621 aaaagaaagt ttgccaaggc acgagtaaca agctcacgca gttgggcact tttgaagatc
     1681 attttctcag cctccagagg atgttcaata actgtgaggt ggtccttggg aatttggaaa
     1741 ttacctatgt gcagaggaat tatgatcttt ccttcttaaa gaccatccag gaggtggctg
     1801 gttatgtcct cattgccctc aacacagtgg agcgaattcc tttggaaaac ctgcagatca
     1861 tcagaggaaa tatgtactac gaaaattcct atgccttagc agtcttatct aactatgatg
     1921 caaataaaac cggactgaag gagctgccca tgagaaattt acaggaaatc ctgcatggcg
     1981 ccgtgcggtt cagcaacaac cctgccctgt gcaacgtgga gagcatccag tggcgggaca
     2041 tagtcagcag tgactttctc agcaacatgt cgatggactt ccagaaccac ctgggcagct
     2101 gccaaaagtg tgatccaagc tgtcccaatg ggagctgctg gggtgcagga gaggagaact
     2161 gccagaaact gaccaaaatc atctgtgccc agcagtgctc cgggtgctgc cgtggcaagt
     2221 cccccagtga ctgctgccac aaccagtgtg ctgcaggctg cacaggcccc cgggagagcg
     2281 actgcctggt ctgccgcaaa ttccgagacg aagccacgtg caaggacacc tgccccccac
     2341 tcatgctcta caaccccacc acgtaccaga tggatgtgaa ccccgagggc aaatacagct
     2401 ttggtgccac ctgcgtgaag aagtgtcccc gtaattatgt ggtgacagat cacggctcgt
     2461 gcgtccgagc ctgtggggcc gacagctatg agatggagga agacggcgtc cgcaagtgta
     2521 agaagtgcga agggccttgc cgcaaagtgt gtaacggaat aggtattggt gaatttaaag
     2581 actcactctc cataaatgct acgaatatta aacacttcaa aaactgcacc tccatcagtg
     2641 gcgatctcca catcctgccg gtggcattta ggggtgactc cttcacacat actcctcctc
     2701 tggatccaca ggaactggat attctgaaaa ccgtaaagga aatcacaggg tttttgctga
     2761 ttcaggcttg gcctgaaaac aggacggacc tccatgcctt tgagaaccta gaaatcatac
     2821 gcggcaggac caagcaacat ggtcagtttt ctcttgcagt cgtcagcctg aacataacat
     2881 ccttgggatt acgctccctc aaggagataa gtgatggaga tgtgataatt tcaggaaaca
     2941 aaaatttgtg ctatgcaaat acaataaact ggaaaaaact gtttgggacc tccggtcaga
     3001 aaaccaaaat tataagcaac agaggtgaaa acagctgcaa ggccacaggc caggtctgcc
     3061 atgccttgtg ctcccccgag ggctgctggg gcccggagcc cagggactgc gtctcttgcc
     3121 ggaatgtcag ccgaggcagg gaatgcgtgg acaagtgcaa ccttctggag ggtgagccaa
     3181 gggagtttgt ggagaactct gagtgcatac agtgccaccc agagtgcctg cctcaggcca
     3241 tgaacatcac ctgcacagga cggggaccag acaactgtat ccagtgtgcc cactacattg
     3301 acggccccca ctgcgtcaag acctgcccgg caggagtcat gggagaaaac aacaccctgg
     3361 tctggaagta cgcagacgcc ggccatgtgt gccacctgtg ccatccaaac tgcacctacg
     3421 gatgcactgg gccaggtctt gaaggctgtc caacgaatgg gcctaagatc ccgtccatcg
     3481 ccactgggat ggtgggggcc ctcctcttgc tgctggtggt ggccctgggg atcggcctct
     3541 tcatgcgaag gcgccacatc gttcggaagc gcacgctgcg gaggctgctg caggagaggg
     3601 agcttgtgga gcctcttaca cccagtggag aagctcccaa ccaagctctc ttgaggatct
     3661 tgaaggaaac tgaattcaaa aagatcaaag tgctgggctc cggtgcgttc ggcacggtgt
     3721 ataagggact ctggatccca gaaggtgaga aagttaaaat tcccgtcgct atcaaggaat
     3781 taagagaagc aacatctccg aaagccaaca aggaaatcct cgatgaagcc tacgtgatgg
     3841 ccagcgtgga caacccccac gtgtgccgcc tgctgggcat ctgcctcacc tccaccgtgc
     3901 agctcatcac gcagctcatg cccttcggct gcctcctgga ctatgtccgg gaacacaaag
     3961 acaatattgg ctcccagtac ctgctcaact ggtgtgtgca gatcgcaaag ggcatgaact
     4021 acttggagga ccgtcgcttg gtgcaccgcg acctggcagc caggaacgta ctggtgaaaa
     4081 caccgcagca tgtcaagatc acagattttg ggctggccaa actgctgggt gcggaagaga
     4141 aagaatacca tgcagaagga ggcaaagtgc ctatcaagtg gatggcattg gaatcaattt
     4201 tacacagaat ctatacccac cagagtgatg tctggagcta cggggtgacc gtttgggagt
     4261 tgatgacctt tggatccaag ccatatgacg gaatccctgc cagcgagatc tcctccatcc
     4321 tggagaaagg agaacgcctc cctcagccac ccatatgtac catcgatgtc tacatgatca
     4381 tggtcaagtg ctggatgata gacgcagata gtcgcccaaa gttccgtgag ttgatcatcg
     4441 aattctccaa aatggcccga gacccccagc gctaccttgt cattcagggg gatgaaagaa
     4501 tgcatttgcc aagtcctaca gactccaact tctaccgtgc cctgatggat gaagaagaca
     4561 tggacgacgt ggtggatgcc gacgagtacc tcatcccaca gcagggcttc ttcagcagcc
     4621 cctccacgtc acggactccc ctcctgagct ctctgagtgc aaccagcaac aattccaccg
     4681 tggcttgcat tgatagaaat gggctgcaaa gctgtcccat caaggaagac agcttcttgc
     4741 agcgatacag ctcagacccc acaggcgcct tgactgagga cagcatagac gacaccttcc
     4801 tcccagtgcc tgaatacata aaccagtccg ttcccaaaag gcccgctggc tctgtgcaga
     4861 atcctgtcta tcacaatcag cctctgaacc ccgcgcccag cagagaccca cactaccagg
     4921 acccccacag cactgcagtg ggcaaccccg agtatctcaa cactgtccag cccacctgtg
     4981 tcaacagcac attcgacagc cctgcccact gggcccagaa aggcagccac caaattagcc
     5041 tggacaaccc tgactaccag caggacttct ttcccaagga agccaagcca aatggcatct
     5101 ttaagggctc cacagctgaa aatgcagaat acctaagggt cgcgccacaa agcagtgaat
     5161 ttattggagc atagaaccca gctttcttgt acaaagtggt gatatccaat taaccctcac
     5221 taaagggaga atgttgaata agaagtaatt ccctatagtg agtcgtatta ccacccaact
     5281 tttctataca aagtggttga tatccagcac agtggcggcc gctcgagtct agagggcccg
     5341 cggttcgaag gtaagcctat ccctaaccct ctcctcggtc tcgattctac gcgtaccggt
     5401 tagtaatgag atccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     5461 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     5521 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     5581 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     5641 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     5701 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     5761 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     5821 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     5881 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     5941 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacac atatgaccga
     6001 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     6061 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     6121 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     6181 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     6241 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     6301 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     6361 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     6421 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     6481 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     6541 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct aaatcgatag
     6601 atcctaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     6661 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     6721 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     6781 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     6841 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     6901 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     6961 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     7021 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     7081 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     7141 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctgaga
     7201 tcctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     7261 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
     7321 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     7381 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     7441 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     7501 cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc
     7561 agagagtgag aggcccgggt taattaagga aagggctaga tcattcttga agacgaaagg
     7621 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     7681 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     7741 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     7801 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     7861 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     7921 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     7981 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     8041 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     8101 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     8161 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     8221 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     8281 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     8341 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     8401 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     8461 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     8521 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     8581 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     8641 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     8701 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     8761 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     8821 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     8881 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     8941 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     9001 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     9061 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     9121 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     9181 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     9241 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     9301 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     9361 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     9421 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     9481 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     9541 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     9601 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     9661 aatgcagcaa gctcatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     9721 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     9781 aaagctcccc gtggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     9841 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     9901 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga
     9961 ttacgaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
    10021 tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc
    10081 ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt catttactct
    10141 aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta
    10201 ctacaaactt agtagttgga agggctaatt cactcccaaa gaagacaaga tatccttgat
    10261 ctgtggatct accacacaca aggctacttc cctgattagc agaactacac accagggcca
    10321 ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat
    10381 aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat
    10441 gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt
    10501 catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc
    10561 tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag
    10621 tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg
    10681 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
    10741 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
    10801 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
    10861 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
    10921 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
    10981 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
    11041 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
    11101 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
    11161 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
    11221 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
    11281 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
    11341 aagaccaccg cacagcaagc ggccggccgc tgatcttcag acctggagga ggagatatga
    11401 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag
    11461 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag
    11521 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga
    11581 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc
    11641 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc
    11701 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt
    11761 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta
    11821 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta
    11881 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga
    11941 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa
    12001 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa
    12061 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat
    12121 cg
//
