LOCUS       sequence-250624-       12145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of EGFR M178I.
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-M178I
FEATURES             Location/Qualifiers
     source          1..12145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    194..311
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        359..1537
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          590..1528
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1485..1505
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1557..1581
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1584..1655,1656..3518,3519..3587,3588..5216)
                     /codon_start=1
                     /product="precursor of human epidermal growth factor 
                     receptor"
                     /label=EGFR
                     /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE
                     DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL
                     QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ
                     WRDIVSSDFLSNMSIDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR
                     CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP
                     EGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI
                     GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK
                     EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD
                     GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG
                     PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP
                     DNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG
                     CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP
                     SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(5218..5242)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5295..5336
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    5357..5924
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5374..5926
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5518..5535)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5745..5764
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5938..5940,5941..5943,5944..6657)
                     /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(5938..5940,5941..5943,5944..6654)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5983..6004)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6244..6263)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6591..6612
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6671..7259
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6724..6744)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7142..7153)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7334..7567
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7686..7704)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7772..7876
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7877..7945,7946..8737)
                     /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      8908..9496
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9397..9416
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9650..9667
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9907..9928
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9943..9948,9949..9966,9967..9973)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9981..9997
                     /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     9986..10008
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10005..10021
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10005..10021
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10071..10090
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             10282..10915
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10962..11087
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11584..11817
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12002..12046
                     /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 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
       61 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
      121 gacagatcca ttcgattagt gaacggatct cgacggtatc gccgaattca caaatggcag
      181 tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
      241 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa
      301 ttcaaaattt tcgggtttat tacagggaca gcagagatcc agtttggact agtcgtgagg
      361 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
      421 aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
      481 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
      541 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt
      601 gtgtggttcc cgcgggcctg gcctctttac gggttatggc ccttgcgtgc cttgaattac
      661 ttccacctgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa gtgggtggga
      721 gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc
      781 tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt
      841 cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt ttttctggca
      901 agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt ttggggccgc
      961 gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc
     1021 gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg
     1081 cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt
     1141 tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa aatggaggac
     1201 gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg cctttccgtc
     1261 ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta
     1321 gttctcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga
     1381 gtttccccac actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt
     1441 ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt
     1501 ggttcaaagt ttttttcttc catttcaggt gtcgtgaagc ggccctgcag atatcaacaa
     1561 gtttgtacaa aaaagcaggc accatgcgac cctccgggac ggccggggca gcgctcctgg
     1621 cgctgctggc tgcgctctgc ccggcgagtc gggctctgga ggaaaagaaa gtttgccaag
     1681 gcacgagtaa caagctcacg cagttgggca cttttgaaga tcattttctc agcctccaga
     1741 ggatgttcaa taactgtgag gtggtccttg ggaatttgga aattacctat gtgcagagga
     1801 attatgatct ttccttctta aagaccatcc aggaggtggc tggttatgtc ctcattgccc
     1861 tcaacacagt ggagcgaatt cctttggaaa acctgcagat catcagagga aatatgtact
     1921 acgaaaattc ctatgcctta gcagtcttat ctaactatga tgcaaataaa accggactga
     1981 aggagctgcc catgagaaat ttacaggaaa tcctgcatgg cgccgtgcgg ttcagcaaca
     2041 accctgccct gtgcaacgtg gagagcatcc agtggcggga catagtcagc agtgactttc
     2101 tcagcaacat gtcgattgac ttccagaacc acctgggcag ctgccaaaag tgtgatccaa
     2161 gctgtcccaa tgggagctgc tggggtgcag gagaggagaa ctgccagaaa ctgaccaaaa
     2221 tcatctgtgc ccagcagtgc tccgggcgct gccgtggcaa gtcccccagt gactgctgcc
     2281 acaaccagtg tgctgcaggc tgcacaggcc cccgggagag cgactgcctg gtctgccgca
     2341 aattccgaga cgaagccacg tgcaaggaca cctgcccccc actcatgctc tacaacccca
     2401 ccacgtacca gatggatgtg aaccccgagg gcaaatacag ctttggtgcc acctgcgtga
     2461 agaagtgtcc ccgtaattat gtggtgacag atcacggctc gtgcgtccga gcctgtgggg
     2521 ccgacagcta tgagatggag gaagacggcg tccgcaagtg taagaagtgc gaagggcctt
     2581 gccgcaaagt gtgtaacgga ataggtattg gtgaatttaa agactcactc tccataaatg
     2641 ctacgaatat taaacacttc aaaaactgca cctccatcag tggcgatctc cacatcctgc
     2701 cggtggcatt taggggtgac tccttcacac atactcctcc tctggatcca caggaactgg
     2761 atattctgaa aaccgtaaag gaaatcacag ggtttttgct gattcaggct tggcctgaaa
     2821 acaggacgga cctccatgcc tttgagaacc tagaaatcat acgcggcagg accaagcaac
     2881 atggtcagtt ttctcttgca gtcgtcagcc tgaacataac atccttggga ttacgctccc
     2941 tcaaggagat aagtgatgga gatgtgataa tttcaggaaa caaaaatttg tgctatgcaa
     3001 atacaataaa ctggaaaaaa ctgtttggga cctccggtca gaaaaccaaa attataagca
     3061 acagaggtga aaacagctgc aaggccacag gccaggtctg ccatgccttg tgctcccccg
     3121 agggctgctg gggcccggag cccagggact gcgtctcttg ccggaatgtc agccgaggca
     3181 gggaatgcgt ggacaagtgc aaccttctgg agggtgagcc aagggagttt gtggagaact
     3241 ctgagtgcat acagtgccac ccagagtgcc tgcctcaggc catgaacatc acctgcacag
     3301 gacggggacc agacaactgt atccagtgtg cccactacat tgacggcccc cactgcgtca
     3361 agacctgccc ggcaggagtc atgggagaaa acaacaccct ggtctggaag tacgcagacg
     3421 ccggccatgt gtgccacctg tgccatccaa actgcaccta cggatgcact gggccaggtc
     3481 ttgaaggctg tccaacgaat gggcctaaga tcccgtccat cgccactggg atggtggggg
     3541 ccctcctctt gctgctggtg gtggccctgg ggatcggcct cttcatgcga aggcgccaca
     3601 tcgttcggaa gcgcacgctg cggaggctgc tgcaggagag ggagcttgtg gagcctctta
     3661 cacccagtgg agaagctccc aaccaagctc tcttgaggat cttgaaggaa actgaattca
     3721 aaaagatcaa agtgctgggc tccggtgcgt tcggcacggt gtataaggga ctctggatcc
     3781 cagaaggtga gaaagttaaa attcccgtcg ctatcaagga attaagagaa gcaacatctc
     3841 cgaaagccaa caaggaaatc ctcgatgaag cctacgtgat ggccagcgtg gacaaccccc
     3901 acgtgtgccg cctgctgggc atctgcctca cctccaccgt gcagctcatc acgcagctca
     3961 tgcccttcgg ctgcctcctg gactatgtcc gggaacacaa agacaatatt ggctcccagt
     4021 acctgctcaa ctggtgtgtg cagatcgcaa agggcatgaa ctacttggag gaccgtcgct
     4081 tggtgcaccg cgacctggca gccaggaacg tactggtgaa aacaccgcag catgtcaaga
     4141 tcacagattt tgggctggcc aaactgctgg gtgcggaaga gaaagaatac catgcagaag
     4201 gaggcaaagt gcctatcaag tggatggcat tggaatcaat tttacacaga atctataccc
     4261 accagagtga tgtctggagc tacggggtga ccgtttggga gttgatgacc tttggatcca
     4321 agccatatga cggaatccct gccagcgaga tctcctccat cctggagaaa ggagaacgcc
     4381 tccctcagcc acccatatgt accatcgatg tctacatgat catggtcaag tgctggatga
     4441 tagacgcaga tagtcgccca aagttccgtg agttgatcat cgaattctcc aaaatggccc
     4501 gagaccccca gcgctacctt gtcattcagg gggatgaaag aatgcatttg ccaagtccta
     4561 cagactccaa cttctaccgt gccctgatgg atgaagaaga catggacgac gtggtggatg
     4621 ccgacgagta cctcatccca cagcagggct tcttcagcag cccctccacg tcacggactc
     4681 ccctcctgag ctctctgagt gcaaccagca acaattccac cgtggcttgc attgatagaa
     4741 atgggctgca aagctgtccc atcaaggaag acagcttctt gcagcgatac agctcagacc
     4801 ccacaggcgc cttgactgag gacagcatag acgacacctt cctcccagtg cctgaataca
     4861 taaaccagtc cgttcccaaa aggcccgctg gctctgtgca gaatcctgtc tatcacaatc
     4921 agcctctgaa ccccgcgccc agcagagacc cacactacca ggacccccac agcactgcag
     4981 tgggcaaccc cgagtatctc aacactgtcc agcccacctg tgtcaacagc acattcgaca
     5041 gccctgccca ctgggcccag aaaggcagcc accaaattag cctggacaac cctgactacc
     5101 agcaggactt ctttcccaag gaagccaagc caaatggcat ctttaagggc tccacagctg
     5161 aaaatgcaga atacctaagg gtcgcgccac aaagcagtga atttattgga gcatagaacc
     5221 cagctttctt gtacaaagtg gttgatatcc agcacagtgg cggccgctcg agtctagagg
     5281 gcccgcggtt cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta
     5341 ccggttagta atgagatccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
     5401 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     5461 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     5521 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     5581 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     5641 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     5701 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     5761 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     5821 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     5881 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacacatatg
     5941 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     6001 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     6061 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     6121 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     6181 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     6241 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     6301 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     6361 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     6421 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     6481 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     6541 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     6601 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaatc
     6661 gatagatcct aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     6721 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     6781 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     6841 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     6901 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     6961 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     7021 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     7081 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     7141 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     7201 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     7261 tgagatcctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     7321 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
     7381 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     7441 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     7501 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     7561 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     7621 atatcagaga gtgagaggcc cgggttaatt aaggaaaggg ctagatcatt cttgaagacg
     7681 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     7741 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     7801 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     7861 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     7921 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     7981 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     8041 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     8101 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     8161 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     8221 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     8281 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     8341 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     8401 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     8461 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     8521 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     8581 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     8641 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     8701 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     8761 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     8821 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     8881 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     8941 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     9001 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     9061 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     9121 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     9181 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     9241 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     9301 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     9361 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     9421 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     9481 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     9541 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     9601 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     9661 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     9721 tcattaatgc agcaagctca tggctgacta atttttttta tttatgcaga ggccgaggcc
     9781 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     9841 tgcaaaaagc tccccgtggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     9901 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     9961 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
    10021 catgattacg aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
    10081 caaactcatc aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa
    10141 tcatcccaaa cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt
    10201 actctaaacc tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat
    10261 atgtactaca aacttagtag ttggaagggc taattcactc ccaaagaaga caagatatcc
    10321 ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag
    10381 ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc
    10441 cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc
    10501 tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag
    10561 catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag
    10621 cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg
    10681 gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct
    10741 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
    10801 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
    10861 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
    10921 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
    10981 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
    11041 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
    11101 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
    11161 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
    11221 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
    11281 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
    11341 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
    11401 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
    11461 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
    11521 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
    11581 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
    11641 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
    11701 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
    11761 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
    11821 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
    11881 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
    11941 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
    12001 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
    12061 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
    12121 tttaagaata gtttttgctg tactt
//
