LOCUS       sequence-250593-       12145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of EGFR L62R.
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-L62R
FEATURES             Location/Qualifiers
     source          1..12145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    101..218
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        266..1444
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          497..1435
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1392..1412
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1464..1488
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1491..1562,1563..3425,3426..3494,3495..5123)
                     /codon_start=1
                     /product="precursor of human epidermal growth factor 
                     receptor"
                     /label=EGFR
                     /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE
                     DHFLSLQRMFNNCEVVRGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL
                     QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ
                     WRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR
                     CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP
                     EGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI
                     GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK
                     EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD
                     GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG
                     PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP
                     DNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG
                     CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP
                     SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(5125..5149)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5202..5243
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    5264..5831
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5281..5833
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5425..5442)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5652..5671
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5845..5847,5848..5850,5851..6564)
                     /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(5845..5847,5848..5850,5851..6561)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5890..5911)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6151..6170)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6498..6519
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6578..7166
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6631..6651)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7049..7060)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7241..7474
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7593..7611)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7679..7783
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7784..7852,7853..8644)
                     /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      8815..9403
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9304..9323
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9557..9574
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9814..9835
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9850..9855,9856..9873,9874..9880)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9888..9904
                     /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     9893..9915
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9912..9928
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9912..9928
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9978..9997
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             10189..10822
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10869..10994
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11491..11724
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11909..11953
                     /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 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
       61 cggtatcgcc gaattcacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
      121 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
      181 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
      241 gagatccagt ttggactagt cgtgaggctc cggtgcccgt cagtgggcag agcgcacatc
      301 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
      361 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
      421 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
      481 tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg
      541 ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga ttcttgatcc
      601 cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc
      661 gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg
      721 gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg
      781 acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca
      841 cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac
      901 atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca
      961 agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc
     1021 ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc
     1081 tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc
     1141 cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta
     1201 ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg
     1261 ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt
     1321 taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc
     1381 ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc
     1441 gtgaagcggc cctgcagata tcaacaagtt tgtacaaaaa agcaggcacc atgcgaccct
     1501 ccgggacggc cggggcagcg ctcctggcgc tgctggctgc gctctgcccg gcgagtcggg
     1561 ctctggagga aaagaaagtt tgccaaggca cgagtaacaa gctcacgcag ttgggcactt
     1621 ttgaagatca ttttctcagc ctccagagga tgttcaataa ctgtgaggtg gtccgtggga
     1681 atttggaaat tacctatgtg cagaggaatt atgatctttc cttcttaaag accatccagg
     1741 aggtggctgg ttatgtcctc attgccctca acacagtgga gcgaattcct ttggaaaacc
     1801 tgcagatcat cagaggaaat atgtactacg aaaattccta tgccttagca gtcttatcta
     1861 actatgatgc aaataaaacc ggactgaagg agctgcccat gagaaattta caggaaatcc
     1921 tgcatggcgc cgtgcggttc agcaacaacc ctgccctgtg caacgtggag agcatccagt
     1981 ggcgggacat agtcagcagt gactttctca gcaacatgtc gatggacttc cagaaccacc
     2041 tgggcagctg ccaaaagtgt gatccaagct gtcccaatgg gagctgctgg ggtgcaggag
     2101 aggagaactg ccagaaactg accaaaatca tctgtgccca gcagtgctcc gggcgctgcc
     2161 gtggcaagtc ccccagtgac tgctgccaca accagtgtgc tgcaggctgc acaggccccc
     2221 gggagagcga ctgcctggtc tgccgcaaat tccgagacga agccacgtgc aaggacacct
     2281 gccccccact catgctctac aaccccacca cgtaccagat ggatgtgaac cccgagggca
     2341 aatacagctt tggtgccacc tgcgtgaaga agtgtccccg taattatgtg gtgacagatc
     2401 acggctcgtg cgtccgagcc tgtggggccg acagctatga gatggaggaa gacggcgtcc
     2461 gcaagtgtaa gaagtgcgaa gggccttgcc gcaaagtgtg taacggaata ggtattggtg
     2521 aatttaaaga ctcactctcc ataaatgcta cgaatattaa acacttcaaa aactgcacct
     2581 ccatcagtgg cgatctccac atcctgccgg tggcatttag gggtgactcc ttcacacata
     2641 ctcctcctct ggatccacag gaactggata ttctgaaaac cgtaaaggaa atcacagggt
     2701 ttttgctgat tcaggcttgg cctgaaaaca ggacggacct ccatgccttt gagaacctag
     2761 aaatcatacg cggcaggacc aagcaacatg gtcagttttc tcttgcagtc gtcagcctga
     2821 acataacatc cttgggatta cgctccctca aggagataag tgatggagat gtgataattt
     2881 caggaaacaa aaatttgtgc tatgcaaata caataaactg gaaaaaactg tttgggacct
     2941 ccggtcagaa aaccaaaatt ataagcaaca gaggtgaaaa cagctgcaag gccacaggcc
     3001 aggtctgcca tgccttgtgc tcccccgagg gctgctgggg cccggagccc agggactgcg
     3061 tctcttgccg gaatgtcagc cgaggcaggg aatgcgtgga caagtgcaac cttctggagg
     3121 gtgagccaag ggagtttgtg gagaactctg agtgcataca gtgccaccca gagtgcctgc
     3181 ctcaggccat gaacatcacc tgcacaggac ggggaccaga caactgtatc cagtgtgccc
     3241 actacattga cggcccccac tgcgtcaaga cctgcccggc aggagtcatg ggagaaaaca
     3301 acaccctggt ctggaagtac gcagacgccg gccatgtgtg ccacctgtgc catccaaact
     3361 gcacctacgg atgcactggg ccaggtcttg aaggctgtcc aacgaatggg cctaagatcc
     3421 cgtccatcgc cactgggatg gtgggggccc tcctcttgct gctggtggtg gccctgggga
     3481 tcggcctctt catgcgaagg cgccacatcg ttcggaagcg cacgctgcgg aggctgctgc
     3541 aggagaggga gcttgtggag cctcttacac ccagtggaga agctcccaac caagctctct
     3601 tgaggatctt gaaggaaact gaattcaaaa agatcaaagt gctgggctcc ggtgcgttcg
     3661 gcacggtgta taagggactc tggatcccag aaggtgagaa agttaaaatt cccgtcgcta
     3721 tcaaggaatt aagagaagca acatctccga aagccaacaa ggaaatcctc gatgaagcct
     3781 acgtgatggc cagcgtggac aacccccacg tgtgccgcct gctgggcatc tgcctcacct
     3841 ccaccgtgca gctcatcacg cagctcatgc ccttcggctg cctcctggac tatgtccggg
     3901 aacacaaaga caatattggc tcccagtacc tgctcaactg gtgtgtgcag atcgcaaagg
     3961 gcatgaacta cttggaggac cgtcgcttgg tgcaccgcga cctggcagcc aggaacgtac
     4021 tggtgaaaac accgcagcat gtcaagatca cagattttgg gctggccaaa ctgctgggtg
     4081 cggaagagaa agaataccat gcagaaggag gcaaagtgcc tatcaagtgg atggcattgg
     4141 aatcaatttt acacagaatc tatacccacc agagtgatgt ctggagctac ggggtgaccg
     4201 tttgggagtt gatgaccttt ggatccaagc catatgacgg aatccctgcc agcgagatct
     4261 cctccatcct ggagaaagga gaacgcctcc ctcagccacc catatgtacc atcgatgtct
     4321 acatgatcat ggtcaagtgc tggatgatag acgcagatag tcgcccaaag ttccgtgagt
     4381 tgatcatcga attctccaaa atggcccgag acccccagcg ctaccttgtc attcaggggg
     4441 atgaaagaat gcatttgcca agtcctacag actccaactt ctaccgtgcc ctgatggatg
     4501 aagaagacat ggacgacgtg gtggatgccg acgagtacct catcccacag cagggcttct
     4561 tcagcagccc ctccacgtca cggactcccc tcctgagctc tctgagtgca accagcaaca
     4621 attccaccgt ggcttgcatt gatagaaatg ggctgcaaag ctgtcccatc aaggaagaca
     4681 gcttcttgca gcgatacagc tcagacccca caggcgcctt gactgaggac agcatagacg
     4741 acaccttcct cccagtgcct gaatacataa accagtccgt tcccaaaagg cccgctggct
     4801 ctgtgcagaa tcctgtctat cacaatcagc ctctgaaccc cgcgcccagc agagacccac
     4861 actaccagga cccccacagc actgcagtgg gcaaccccga gtatctcaac actgtccagc
     4921 ccacctgtgt caacagcaca ttcgacagcc ctgcccactg ggcccagaaa ggcagccacc
     4981 aaattagcct ggacaaccct gactaccagc aggacttctt tcccaaggaa gccaagccaa
     5041 atggcatctt taagggctcc acagctgaaa atgcagaata cctaagggtc gcgccacaaa
     5101 gcagtgaatt tattggagca tagaacccag ctttcttgta caaagtggtt gatatccagc
     5161 acagtggcgg ccgctcgagt ctagagggcc cgcggttcga aggtaagcct atccctaacc
     5221 ctctcctcgg tctcgattct acgcgtaccg gttagtaatg agatccctcc ccccccccta
     5281 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     5341 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     5401 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     5461 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     5521 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     5581 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     5641 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     5701 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     5761 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     5821 cacgatgata atatggccac acatatggtg agcaagggcg aggagctgtt caccggggtg
     5881 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     5941 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     6001 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     6061 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     6121 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     6181 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     6241 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     6301 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     6361 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     6421 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     6481 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     6541 ggcatggacg agctgtacaa gtaaatcgat agatcctaat caacctctgg attacaaaat
     6601 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     6661 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     6721 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     6781 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     6841 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     6901 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     6961 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     7021 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     7081 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     7141 gatctccctt tgggccgcct ccccgcctga gatcctttaa gaccaatgac ttacaaggca
     7201 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     7261 caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     7321 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     7381 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     7441 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
     7501 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggcccgg gttaattaag
     7561 gaaagggcta gatcattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     7621 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     7681 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     7741 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     7801 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     7861 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     7921 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     7981 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     8041 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     8101 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     8161 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     8221 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     8281 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     8341 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     8401 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     8461 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     8521 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     8581 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     8641 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     8701 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     8761 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     8821 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     8881 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     8941 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     9001 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     9061 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     9121 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     9181 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     9241 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     9301 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     9361 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     9421 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     9481 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     9541 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     9601 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc aagctcatgg ctgactaatt
     9661 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     9721 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc ccgtggcacg acaggtttcc
     9781 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     9841 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     9901 acaatttcac acaggaaaca gctatgacat gattacgaat ttcacaaata aagcattttt
     9961 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
    10021 caactggata actcaagcta accaaaatca tcccaaactt cccaccccat accctattac
    10081 cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg aattattttc
    10141 attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagttg gaagggctaa
    10201 ttcactccca aagaagacaa gatatccttg atctgtggat ctaccacaca caaggctact
    10261 tccctgatta gcagaactac acaccagggc caggggtcag atatccactg acctttggat
    10321 ggtgctacaa gctagtacca gttgagccag ataaggtaga agaggccaat aaaggagaga
    10381 acaccagctt gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtgt
    10441 tagagtggag gtttgacagc cgcctagcat ttcatcacgt ggcccgagag ctgcatccgg
    10501 agtacttcaa gaactgctga tatcgagctt gctacaaggg actttccgct ggggactttc
    10561 cagggaggcg tggcctgggc gggactgggg agtggcgagc cctcagatcc tgcatataag
    10621 cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
    10681 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
    10741 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
    10801 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
    10861 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
    10921 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
    10981 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
    11041 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
    11101 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
    11161 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
    11221 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
    11281 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccggcc
    11341 gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa
    11401 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
    11461 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
    11521 agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt
    11581 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
    11641 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag
    11701 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
    11761 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca
    11821 cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt
    11881 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa
    11941 atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt
    12001 cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt
    12061 gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag
    12121 gggacccgac aggcccgaag gaata
//
