LOCUS       sequence-250615-       12122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of EGFR L838V.
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-L838V
FEATURES             Location/Qualifiers
     source          1..12122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    127..244
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        292..1470
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          523..1461
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1418..1438
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1490..1514
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1517..1588,1589..3451,3452..3520,3521..5149)
                     /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
                     SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDVAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(5151..5175)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        5183..5201
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5226..5245)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5227..5245)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(5251..5271)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             5325..5366
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    5387..5954
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5404..5956
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5548..5565)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5775..5794
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5968..6567
                     /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(5968..5987)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6464..6484
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6581..7169
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6634..6654)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7052..7063)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7244..7477
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7596..7614)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7682..7786
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7787..7855,7856..8647)
                     /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      8818..9406
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9307..9326
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9560..9577
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9817..9838
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9853..9858,9859..9876,9877..9883)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9891..9907
                     /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     9896..9918
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9915..9931
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9915..9931
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9981..10000
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             10192..10825
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10872..10997
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11494..11727
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11912..11956
                     /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 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
       61 ccattcgatt agtgaacgga tctcgacggt atcgccgaat tcacaaatgg cagtattcat
      121 ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
      181 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
      241 ttttcgggtt tattacaggg acagcagaga tccagtttgg actagtcgtg aggctccggt
      301 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
      361 ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
      421 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
      481 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt
      541 tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc
      601 tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg
      661 aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc
      721 tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag
      781 tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt
      841 cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc
      901 gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca
      961 ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg
     1021 ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga
     1081 gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc
     1141 tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc
     1201 gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg
     1261 agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc
     1321 cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg
     1381 gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa
     1441 agtttttttc ttccatttca ggtgtcgtga agcggccctg cagatatcaa caagtttgta
     1501 caaaaaagca ggcaccatgc gaccctccgg gacggccggg gcagcgctcc tggcgctgct
     1561 ggctgcgctc tgcccggcga gtcgggctct ggaggaaaag aaagtttgcc aaggcacgag
     1621 taacaagctc acgcagttgg gcacttttga agatcatttt ctcagcctcc agaggatgtt
     1681 caataactgt gaggtggtcc ttgggaattt ggaaattacc tatgtgcaga ggaattatga
     1741 tctttccttc ttaaagacca tccaggaggt ggctggttat gtcctcattg ccctcaacac
     1801 agtggagcga attcctttgg aaaacctgca gatcatcaga ggaaatatgt actacgaaaa
     1861 ttcctatgcc ttagcagtct tatctaacta tgatgcaaat aaaaccggac tgaaggagct
     1921 gcccatgaga aatttacagg aaatcctgca tggcgccgtg cggttcagca acaaccctgc
     1981 cctgtgcaac gtggagagca tccagtggcg ggacatagtc agcagtgact ttctcagcaa
     2041 catgtcgatg gacttccaga accacctggg cagctgccaa aagtgtgatc caagctgtcc
     2101 caatgggagc tgctggggtg caggagagga gaactgccag aaactgacca aaatcatctg
     2161 tgcccagcag tgctccgggc gctgccgtgg caagtccccc agtgactgct gccacaacca
     2221 gtgtgctgca ggctgcacag gcccccggga gagcgactgc ctggtctgcc gcaaattccg
     2281 agacgaagcc acgtgcaagg acacctgccc cccactcatg ctctacaacc ccaccacgta
     2341 ccagatggat gtgaaccccg agggcaaata cagctttggt gccacctgcg tgaagaagtg
     2401 tccccgtaat tatgtggtga cagatcacgg ctcgtgcgtc cgagcctgtg gggccgacag
     2461 ctatgagatg gaggaagacg gcgtccgcaa gtgtaagaag tgcgaagggc cttgccgcaa
     2521 agtgtgtaac ggaataggta ttggtgaatt taaagactca ctctccataa atgctacgaa
     2581 tattaaacac ttcaaaaact gcacctccat cagtggcgat ctccacatcc tgccggtggc
     2641 atttaggggt gactccttca cacatactcc tcctctggat ccacaggaac tggatattct
     2701 gaaaaccgta aaggaaatca cagggttttt gctgattcag gcttggcctg aaaacaggac
     2761 ggacctccat gcctttgaga acctagaaat catacgcggc aggaccaagc aacatggtca
     2821 gttttctctt gcagtcgtca gcctgaacat aacatccttg ggattacgct ccctcaagga
     2881 gataagtgat ggagatgtga taatttcagg aaacaaaaat ttgtgctatg caaatacaat
     2941 aaactggaaa aaactgtttg ggacctccgg tcagaaaacc aaaattataa gcaacagagg
     3001 tgaaaacagc tgcaaggcca caggccaggt ctgccatgcc ttgtgctccc ccgagggctg
     3061 ctggggcccg gagcccaggg actgcgtctc ttgccggaat gtcagccgag gcagggaatg
     3121 cgtggacaag tgcaaccttc tggagggtga gccaagggag tttgtggaga actctgagtg
     3181 catacagtgc cacccagagt gcctgcctca ggccatgaac atcacctgca caggacgggg
     3241 accagacaac tgtatccagt gtgcccacta cattgacggc ccccactgcg tcaagacctg
     3301 cccggcagga gtcatgggag aaaacaacac cctggtctgg aagtacgcag acgccggcca
     3361 tgtgtgccac ctgtgccatc caaactgcac ctacggatgc actgggccag gtcttgaagg
     3421 ctgtccaacg aatgggccta agatcccgtc catcgccact gggatggtgg gggccctcct
     3481 cttgctgctg gtggtggccc tggggatcgg cctcttcatg cgaaggcgcc acatcgttcg
     3541 gaagcgcacg ctgcggaggc tgctgcagga gagggagctt gtggagcctc ttacacccag
     3601 tggagaagct cccaaccaag ctctcttgag gatcttgaag gaaactgaat tcaaaaagat
     3661 caaagtgctg ggctccggtg cgttcggcac ggtgtataag ggactctgga tcccagaagg
     3721 tgagaaagtt aaaattcccg tcgctatcaa ggaattaaga gaagcaacat ctccgaaagc
     3781 caacaaggaa atcctcgatg aagcctacgt gatggccagc gtggacaacc cccacgtgtg
     3841 ccgcctgctg ggcatctgcc tcacctccac cgtgcagctc atcacgcagc tcatgccctt
     3901 cggctgcctc ctggactatg tccgggaaca caaagacaat attggctccc agtacctgct
     3961 caactggtgt gtgcagatcg caaagggcat gaactacttg gaggaccgtc gcttggtgca
     4021 ccgcgacgtg gcagccagga acgtactggt gaaaacaccg cagcatgtca agatcacaga
     4081 ttttgggctg gccaaactgc tgggtgcgga agagaaagaa taccatgcag aaggaggcaa
     4141 agtgcctatc aagtggatgg cattggaatc aattttacac agaatctata cccaccagag
     4201 tgatgtctgg agctacgggg tgaccgtttg ggagttgatg acctttggat ccaagccata
     4261 tgacggaatc cctgccagcg agatctcctc catcctggag aaaggagaac gcctccctca
     4321 gccacccata tgtaccatcg atgtctacat gatcatggtc aagtgctgga tgatagacgc
     4381 agatagtcgc ccaaagttcc gtgagttgat catcgaattc tccaaaatgg cccgagaccc
     4441 ccagcgctac cttgtcattc agggggatga aagaatgcat ttgccaagtc ctacagactc
     4501 caacttctac cgtgccctga tggatgaaga agacatggac gacgtggtgg atgccgacga
     4561 gtacctcatc ccacagcagg gcttcttcag cagcccctcc acgtcacgga ctcccctcct
     4621 gagctctctg agtgcaacca gcaacaattc caccgtggct tgcattgata gaaatgggct
     4681 gcaaagctgt cccatcaagg aagacagctt cttgcagcga tacagctcag accccacagg
     4741 cgccttgact gaggacagca tagacgacac cttcctccca gtgcctgaat acataaacca
     4801 gtccgttccc aaaaggcccg ctggctctgt gcagaatcct gtctatcaca atcagcctct
     4861 gaaccccgcg cccagcagag acccacacta ccaggacccc cacagcactg cagtgggcaa
     4921 ccccgagtat ctcaacactg tccagcccac ctgtgtcaac agcacattcg acagccctgc
     4981 ccactgggcc cagaaaggca gccaccaaat tagcctggac aaccctgact accagcagga
     5041 cttctttccc aaggaagcca agccaaatgg catctttaag ggctccacag ctgaaaatgc
     5101 agaataccta agggtcgcgc cacaaagcag tgaatttatt ggagcataga acccagcttt
     5161 cttgtacaaa gtggtgatat ccaattaacc ctcactaaag gtttagagtt tgattagtat
     5221 gtttgcccta tagtgagtcg tattaccacc caacttttct atacaaagtg gttgatatcc
     5281 agcacagtgg cggccgctcg agtctagagg gcccgcggtt cgaaggtaag cctatcccta
     5341 accctctcct cggtctcgat tctacgcgta ccggttagta atgagatccc tccccccccc
     5401 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     5461 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     5521 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     5581 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc
     5641 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     5701 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     5761 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     5821 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt
     5881 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     5941 aaacacgatg ataatatggc cacacatatg accgagtaca agcccacggt gcgcctcgcc
     6001 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     6061 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     6121 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     6181 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     6241 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     6301 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     6361 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     6421 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     6481 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     6541 tgcatgaccc gcaagcccgg tgcctaaatc gatagatcct aatcaacctc tggattacaa
     6601 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     6661 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     6721 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     6781 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     6841 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     6901 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     6961 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     7021 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     7081 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     7141 tcggatctcc ctttgggccg cctccccgcc tgagatcctt taagaccaat gacttacaag
     7201 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     7261 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     7321 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     7381 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     7441 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     7501 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggcc cgggttaatt
     7561 aaggaaaggg ctagatcatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     7621 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     7681 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     7741 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     7801 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     7861 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     7921 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     7981 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     8041 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     8101 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     8161 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     8221 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     8281 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     8341 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     8401 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     8461 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     8521 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     8581 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     8641 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     8701 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     8761 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     8821 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     8881 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     8941 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     9001 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     9061 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     9121 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     9181 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     9241 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     9301 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     9361 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     9421 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     9481 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     9541 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     9601 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agcaagctca tggctgacta
     9661 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     9721 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tccccgtggc acgacaggtt
     9781 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     9841 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     9901 ataacaattt cacacaggaa acagctatga catgattacg aatttcacaa ataaagcatt
     9961 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
    10021 gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat
    10081 taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt
    10141 ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag ttggaagggc
    10201 taattcactc ccaaagaaga caagatatcc ttgatctgtg gatctaccac acacaaggct
    10261 acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca ctgacctttg
    10321 gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc aataaaggag
    10381 agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag
    10441 tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga gagctgcatc
    10501 cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc gctggggact
    10561 ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga tcctgcatat
    10621 aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg agcctgggag
    10681 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
    10741 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
    10801 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
    10861 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
    10921 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
    10981 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
    11041 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
    11101 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
    11161 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
    11221 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
    11281 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
    11341 gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata
    11401 taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag
    11461 agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg
    11521 agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt
    11581 attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca
    11641 tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga
    11701 aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg
    11761 caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa
    11821 tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct taatacactc
    11881 cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga
    11941 taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt
    12001 attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat
    12061 agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc
    12121 ga
//
