LOCUS       sequence-250499-       12145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of EGFR E114K.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12145)
  AUTHORS   Ng PK, Li J, Jeong KJ, Shao S, Chen H, Tsang YH, Sengupta S, Wang Z,
            Bhavana VH, Tran R, Soewito S, Minussi DC, Moreno D, Kong K, 
            Dogruluk T, Lu H, Gao J, Tokheim C, Zhou DC, Johnson AM, Zeng J, Ip 
            CKM, Ju Z, Wester M, Yu S, Li Y, Vellano CP, Schultz N, Karchin R, 
            Ding L, Lu Y, Cheung LWT, Chen K, Shaw KR, Meric-Bernstam F, Scott 
            KL, Yi S, Sahni N, Liang H, Mills GB
  TITLE     Systematic Functional Annotation of Somatic Mutations in Cancer.
  JOURNAL   Cancer Cell. 2018 Mar 12;33(3):450-462.e10. doi: 
            10.1016/j.ccell.2018.01.021.
  PUBMED    29533785
REFERENCE   2  (bases 1 to 12145)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.ccell.2018.01"; journalName: "Cancer Cell"; date: 
            "2018-03-12- 12"; volume: "33"; issue: "3"; pages: "450-462.e10"
COMMENT     Sequence Label: pHAGE-EGFR-E114K
FEATURES             Location/Qualifiers
     source          1..12145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             41..85
                     /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"
     misc_feature    378..495
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        543..1721
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          774..1712
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1669..1689
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1741..1765
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1768..1839,1840..3702,3703..3771,3772..5400)
                     /codon_start=1
                     /product="precursor of human epidermal growth factor 
                     receptor"
                     /label=EGFR
                     /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE
                     DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL
                     QIIRGNMYYKNSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ
                     WRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR
                     CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP
                     EGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI
                     GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK
                     EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD
                     GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG
                     PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP
                     DNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG
                     CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP
                     SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(5402..5426)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5479..5520
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    5541..6108
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5558..6110
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5702..5719)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5929..5948
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(6122..6124,6125..6127,6128..6841)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6122..6124,6125..6127,6128..6838)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6167..6188)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6428..6447)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6775..6796
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6855..7443
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6908..6928)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7326..7337)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7518..7751
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7870..7888)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7956..8060
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8061..8129,8130..8921)
                     /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      9092..9680
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9581..9600
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9834..9851
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10091..10112
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10127..10132,10133..10150,10151..10157)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10165..10181
                     /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     10170..10192
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10189..10205
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10189..10205
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10255..10274
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             10466..11099
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    11146..11271
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11768..12001
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
       61 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
      121 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
      181 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
      241 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
      301 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
      361 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
      421 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
      481 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcgt
      541 gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
      601 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
      661 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
      721 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
      781 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
      841 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
      901 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
      961 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
     1021 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
     1081 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
     1141 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
     1201 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
     1261 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     1321 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     1381 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     1441 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     1501 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     1561 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     1621 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     1681 cagtggttca aagttttttt cttccatttc aggtgtcgtg aagcggccct gcagatatca
     1741 acaagtttgt acaaaaaagc aggcaccatg cgaccctccg ggacggccgg ggcagcgctc
     1801 ctggcgctgc tggctgcgct ctgcccggcg agtcgggctc tggaggaaaa gaaagtttgc
     1861 caaggcacga gtaacaagct cacgcagttg ggcacttttg aagatcattt tctcagcctc
     1921 cagaggatgt tcaataactg tgaggtggtc cttgggaatt tggaaattac ctatgtgcag
     1981 aggaattatg atctttcctt cttaaagacc atccaggagg tggctggtta tgtcctcatt
     2041 gccctcaaca cagtggagcg aattcctttg gaaaacctgc agatcatcag aggaaatatg
     2101 tactacaaaa attcctatgc cttagcagtc ttatctaact atgatgcaaa taaaaccgga
     2161 ctgaaggagc tgcccatgag aaatttacag gaaatcctgc atggcgccgt gcggttcagc
     2221 aacaaccctg ccctgtgcaa cgtggagagc atccagtggc gggacatagt cagcagtgac
     2281 tttctcagca acatgtcgat ggacttccag aaccacctgg gcagctgcca aaagtgtgat
     2341 ccaagctgtc ccaatgggag ctgctggggt gcaggagagg agaactgcca gaaactgacc
     2401 aaaatcatct gtgcccagca gtgctccggg cgctgccgtg gcaagtcccc cagtgactgc
     2461 tgccacaacc agtgtgctgc aggctgcaca ggcccccggg agagcgactg cctggtctgc
     2521 cgcaaattcc gagacgaagc cacgtgcaag gacacctgcc ccccactcat gctctacaac
     2581 cccaccacgt accagatgga tgtgaacccc gagggcaaat acagctttgg tgccacctgc
     2641 gtgaagaagt gtccccgtaa ttatgtggtg acagatcacg gctcgtgcgt ccgagcctgt
     2701 ggggccgaca gctatgagat ggaggaagac ggcgtccgca agtgtaagaa gtgcgaaggg
     2761 ccttgccgca aagtgtgtaa cggaataggt attggtgaat ttaaagactc actctccata
     2821 aatgctacga atattaaaca cttcaaaaac tgcacctcca tcagtggcga tctccacatc
     2881 ctgccggtgg catttagggg tgactccttc acacatactc ctcctctgga tccacaggaa
     2941 ctggatattc tgaaaaccgt aaaggaaatc acagggtttt tgctgattca ggcttggcct
     3001 gaaaacagga cggacctcca tgcctttgag aacctagaaa tcatacgcgg caggaccaag
     3061 caacatggtc agttttctct tgcagtcgtc agcctgaaca taacatcctt gggattacgc
     3121 tccctcaagg agataagtga tggagatgtg ataatttcag gaaacaaaaa tttgtgctat
     3181 gcaaatacaa taaactggaa aaaactgttt gggacctccg gtcagaaaac caaaattata
     3241 agcaacagag gtgaaaacag ctgcaaggcc acaggccagg tctgccatgc cttgtgctcc
     3301 cccgagggct gctggggccc ggagcccagg gactgcgtct cttgccggaa tgtcagccga
     3361 ggcagggaat gcgtggacaa gtgcaacctt ctggagggtg agccaaggga gtttgtggag
     3421 aactctgagt gcatacagtg ccacccagag tgcctgcctc aggccatgaa catcacctgc
     3481 acaggacggg gaccagacaa ctgtatccag tgtgcccact acattgacgg cccccactgc
     3541 gtcaagacct gcccggcagg agtcatggga gaaaacaaca ccctggtctg gaagtacgca
     3601 gacgccggcc atgtgtgcca cctgtgccat ccaaactgca cctacggatg cactgggcca
     3661 ggtcttgaag gctgtccaac gaatgggcct aagatcccgt ccatcgccac tgggatggtg
     3721 ggggccctcc tcttgctgct ggtggtggcc ctggggatcg gcctcttcat gcgaaggcgc
     3781 cacatcgttc ggaagcgcac gctgcggagg ctgctgcagg agagggagct tgtggagcct
     3841 cttacaccca gtggagaagc tcccaaccaa gctctcttga ggatcttgaa ggaaactgaa
     3901 ttcaaaaaga tcaaagtgct gggctccggt gcgttcggca cggtgtataa gggactctgg
     3961 atcccagaag gtgagaaagt taaaattccc gtcgctatca aggaattaag agaagcaaca
     4021 tctccgaaag ccaacaagga aatcctcgat gaagcctacg tgatggccag cgtggacaac
     4081 ccccacgtgt gccgcctgct gggcatctgc ctcacctcca ccgtgcagct catcacgcag
     4141 ctcatgccct tcggctgcct cctggactat gtccgggaac acaaagacaa tattggctcc
     4201 cagtacctgc tcaactggtg tgtgcagatc gcaaagggca tgaactactt ggaggaccgt
     4261 cgcttggtgc accgcgacct ggcagccagg aacgtactgg tgaaaacacc gcagcatgtc
     4321 aagatcacag attttgggct ggccaaactg ctgggtgcgg aagagaaaga ataccatgca
     4381 gaaggaggca aagtgcctat caagtggatg gcattggaat caattttaca cagaatctat
     4441 acccaccaga gtgatgtctg gagctacggg gtgaccgttt gggagttgat gacctttgga
     4501 tccaagccat atgacggaat ccctgccagc gagatctcct ccatcctgga gaaaggagaa
     4561 cgcctccctc agccacccat atgtaccatc gatgtctaca tgatcatggt caagtgctgg
     4621 atgatagacg cagatagtcg cccaaagttc cgtgagttga tcatcgaatt ctccaaaatg
     4681 gcccgagacc cccagcgcta ccttgtcatt cagggggatg aaagaatgca tttgccaagt
     4741 cctacagact ccaacttcta ccgtgccctg atggatgaag aagacatgga cgacgtggtg
     4801 gatgccgacg agtacctcat cccacagcag ggcttcttca gcagcccctc cacgtcacgg
     4861 actcccctcc tgagctctct gagtgcaacc agcaacaatt ccaccgtggc ttgcattgat
     4921 agaaatgggc tgcaaagctg tcccatcaag gaagacagct tcttgcagcg atacagctca
     4981 gaccccacag gcgccttgac tgaggacagc atagacgaca ccttcctccc agtgcctgaa
     5041 tacataaacc agtccgttcc caaaaggccc gctggctctg tgcagaatcc tgtctatcac
     5101 aatcagcctc tgaaccccgc gcccagcaga gacccacact accaggaccc ccacagcact
     5161 gcagtgggca accccgagta tctcaacact gtccagccca cctgtgtcaa cagcacattc
     5221 gacagccctg cccactgggc ccagaaaggc agccaccaaa ttagcctgga caaccctgac
     5281 taccagcagg acttctttcc caaggaagcc aagccaaatg gcatctttaa gggctccaca
     5341 gctgaaaatg cagaatacct aagggtcgcg ccacaaagca gtgaatttat tggagcatag
     5401 aacccagctt tcttgtacaa agtggttgat atccagcaca gtggcggccg ctcgagtcta
     5461 gagggcccgc ggttcgaagg taagcctatc cctaaccctc tcctcggtct cgattctacg
     5521 cgtaccggtt agtaatgaga tccctccccc ccccctaacg ttactggccg aagccgcttg
     5581 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc
     5641 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc
     5701 cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa
     5761 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct
     5821 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca
     5881 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca
     5941 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat
     6001 ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc
     6061 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaca
     6121 tatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     6181 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     6241 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     6301 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     6361 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     6421 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     6481 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     6541 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     6601 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     6661 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     6721 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     6781 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     6841 aatcgataga tcctaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     6901 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     6961 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     7021 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     7081 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     7141 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     7201 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     7261 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     7321 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     7381 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     7441 cgcctgagat cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     7501 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt
     7561 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     7621 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     7681 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     7741 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     7801 atgaatatca gagagtgaga ggcccgggtt aattaaggaa agggctagat cattcttgaa
     7861 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     7921 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     7981 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     8041 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     8101 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     8161 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     8221 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     8281 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     8341 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     8401 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     8461 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     8521 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     8581 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     8641 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     8701 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     8761 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     8821 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     8881 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     8941 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     9001 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     9061 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     9121 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     9181 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     9241 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     9301 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     9361 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     9421 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     9481 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     9541 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     9601 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     9661 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     9721 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     9781 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     9841 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     9901 cgattcatta atgcagcaag ctcatggctg actaattttt tttatttatg cagaggccga
     9961 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
    10021 cttttgcaaa aagctccccg tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
    10081 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
    10141 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
    10201 atgacatgat tacgaatttc acaaataaag catttttttc actgcattct agttgtggtt
    10261 tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc
    10321 aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac ctgtggtttc
    10381 atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat tgtatttgtt
    10441 aaatatgtac tacaaactta gtagttggaa gggctaattc actcccaaag aagacaagat
    10501 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca
    10561 ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt
    10621 gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg
    10681 agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc
    10741 ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat
    10801 cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg
    10861 actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg
    10921 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
    10981 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
    11041 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
    11101 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
    11161 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
    11221 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
    11281 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
    11341 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
    11401 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
    11461 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
    11521 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
    11581 aacaaaagta agaccaccgc acagcaagcg gccggccgct gatcttcaga cctggaggag
    11641 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
    11701 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
    11761 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
    11821 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
    11881 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
    11941 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
    12001 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
    12061 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
    12121 gagaaattaa caattacaca agctt
//
