LOCUS       sequence-250522-       12145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of EGFR E967A.
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-E967A
FEATURES             Location/Qualifiers
     source          1..12145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             34..78
                     /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    371..488
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        536..1714
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          767..1705
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1662..1682
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1734..1758
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1761..1832,1833..3695,3696..3764,3765..5393)
                     /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
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIAFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(5395..5419)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5472..5513
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    5534..6101
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5551..6103
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5695..5712)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5922..5941
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(6115..6117,6118..6120,6121..6834)
                     /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(6115..6117,6118..6120,6121..6831)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6160..6181)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6421..6440)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6768..6789
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6848..7436
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6901..6921)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7319..7330)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7511..7744
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7863..7881)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7949..8053
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8054..8122,8123..8914)
                     /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      9085..9673
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9574..9593
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9827..9844
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10084..10105
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10120..10125,10126..10143,10144..10150)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10158..10174
                     /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     10163..10185
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10182..10198
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10182..10198
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10248..10267
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             10459..11092
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    11139..11264
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11761..11994
                     /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 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
       61 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
      121 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
      181 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
      241 ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac
      301 agatccattc gattagtgaa cggatctcga cggtatcgcc gaattcacaa atggcagtat
      361 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
      421 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
      481 aaaattttcg ggtttattac agggacagca gagatccagt ttggactagt cgtgaggctc
      541 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
      601 ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
      661 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
      721 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg
      781 tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc
      841 cacctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag
      901 ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg
      961 gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga
     1021 taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga
     1081 tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg
     1141 cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg
     1201 gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct
     1261 cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc
     1321 gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg
     1381 gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc
     1441 agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt
     1501 ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt
     1561 tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc
     1621 cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt
     1681 tcaaagtttt tttcttccat ttcaggtgtc gtgaagcggc cctgcagata tcaacaagtt
     1741 tgtacaaaaa agcaggcacc atgcgaccct ccgggacggc cggggcagcg ctcctggcgc
     1801 tgctggctgc gctctgcccg gcgagtcggg ctctggagga aaagaaagtt tgccaaggca
     1861 cgagtaacaa gctcacgcag ttgggcactt ttgaagatca ttttctcagc ctccagagga
     1921 tgttcaataa ctgtgaggtg gtccttggga atttggaaat tacctatgtg cagaggaatt
     1981 atgatctttc cttcttaaag accatccagg aggtggctgg ttatgtcctc attgccctca
     2041 acacagtgga gcgaattcct ttggaaaacc tgcagatcat cagaggaaat atgtactacg
     2101 aaaattccta tgccttagca gtcttatcta actatgatgc aaataaaacc ggactgaagg
     2161 agctgcccat gagaaattta caggaaatcc tgcatggcgc cgtgcggttc agcaacaacc
     2221 ctgccctgtg caacgtggag agcatccagt ggcgggacat agtcagcagt gactttctca
     2281 gcaacatgtc gatggacttc cagaaccacc tgggcagctg ccaaaagtgt gatccaagct
     2341 gtcccaatgg gagctgctgg ggtgcaggag aggagaactg ccagaaactg accaaaatca
     2401 tctgtgccca gcagtgctcc gggcgctgcc gtggcaagtc ccccagtgac tgctgccaca
     2461 accagtgtgc tgcaggctgc acaggccccc gggagagcga ctgcctggtc tgccgcaaat
     2521 tccgagacga agccacgtgc aaggacacct gccccccact catgctctac aaccccacca
     2581 cgtaccagat ggatgtgaac cccgagggca aatacagctt tggtgccacc tgcgtgaaga
     2641 agtgtccccg taattatgtg gtgacagatc acggctcgtg cgtccgagcc tgtggggccg
     2701 acagctatga gatggaggaa gacggcgtcc gcaagtgtaa gaagtgcgaa gggccttgcc
     2761 gcaaagtgtg taacggaata ggtattggtg aatttaaaga ctcactctcc ataaatgcta
     2821 cgaatattaa acacttcaaa aactgcacct ccatcagtgg cgatctccac atcctgccgg
     2881 tggcatttag gggtgactcc ttcacacata ctcctcctct ggatccacag gaactggata
     2941 ttctgaaaac cgtaaaggaa atcacagggt ttttgctgat tcaggcttgg cctgaaaaca
     3001 ggacggacct ccatgccttt gagaacctag aaatcatacg cggcaggacc aagcaacatg
     3061 gtcagttttc tcttgcagtc gtcagcctga acataacatc cttgggatta cgctccctca
     3121 aggagataag tgatggagat gtgataattt caggaaacaa aaatttgtgc tatgcaaata
     3181 caataaactg gaaaaaactg tttgggacct ccggtcagaa aaccaaaatt ataagcaaca
     3241 gaggtgaaaa cagctgcaag gccacaggcc aggtctgcca tgccttgtgc tcccccgagg
     3301 gctgctgggg cccggagccc agggactgcg tctcttgccg gaatgtcagc cgaggcaggg
     3361 aatgcgtgga caagtgcaac cttctggagg gtgagccaag ggagtttgtg gagaactctg
     3421 agtgcataca gtgccaccca gagtgcctgc ctcaggccat gaacatcacc tgcacaggac
     3481 ggggaccaga caactgtatc cagtgtgccc actacattga cggcccccac tgcgtcaaga
     3541 cctgcccggc aggagtcatg ggagaaaaca acaccctggt ctggaagtac gcagacgccg
     3601 gccatgtgtg ccacctgtgc catccaaact gcacctacgg atgcactggg ccaggtcttg
     3661 aaggctgtcc aacgaatggg cctaagatcc cgtccatcgc cactgggatg gtgggggccc
     3721 tcctcttgct gctggtggtg gccctgggga tcggcctctt catgcgaagg cgccacatcg
     3781 ttcggaagcg cacgctgcgg aggctgctgc aggagaggga gcttgtggag cctcttacac
     3841 ccagtggaga agctcccaac caagctctct tgaggatctt gaaggaaact gaattcaaaa
     3901 agatcaaagt gctgggctcc ggtgcgttcg gcacggtgta taagggactc tggatcccag
     3961 aaggtgagaa agttaaaatt cccgtcgcta tcaaggaatt aagagaagca acatctccga
     4021 aagccaacaa ggaaatcctc gatgaagcct acgtgatggc cagcgtggac aacccccacg
     4081 tgtgccgcct gctgggcatc tgcctcacct ccaccgtgca gctcatcacg cagctcatgc
     4141 ccttcggctg cctcctggac tatgtccggg aacacaaaga caatattggc tcccagtacc
     4201 tgctcaactg gtgtgtgcag atcgcaaagg gcatgaacta cttggaggac cgtcgcttgg
     4261 tgcaccgcga cctggcagcc aggaacgtac tggtgaaaac accgcagcat gtcaagatca
     4321 cagattttgg gctggccaaa ctgctgggtg cggaagagaa agaataccat gcagaaggag
     4381 gcaaagtgcc tatcaagtgg atggcattgg aatcaatttt acacagaatc tatacccacc
     4441 agagtgatgt ctggagctac ggggtgaccg tttgggagtt gatgaccttt ggatccaagc
     4501 catatgacgg aatccctgcc agcgagatct cctccatcct ggagaaagga gaacgcctcc
     4561 ctcagccacc catatgtacc atcgatgtct acatgatcat ggtcaagtgc tggatgatag
     4621 acgcagatag tcgcccaaag ttccgtgagt tgatcatcgc attctccaaa atggcccgag
     4681 acccccagcg ctaccttgtc attcaggggg atgaaagaat gcatttgcca agtcctacag
     4741 actccaactt ctaccgtgcc ctgatggatg aagaagacat ggacgacgtg gtggatgccg
     4801 acgagtacct catcccacag cagggcttct tcagcagccc ctccacgtca cggactcccc
     4861 tcctgagctc tctgagtgca accagcaaca attccaccgt ggcttgcatt gatagaaatg
     4921 ggctgcaaag ctgtcccatc aaggaagaca gcttcttgca gcgatacagc tcagacccca
     4981 caggcgcctt gactgaggac agcatagacg acaccttcct cccagtgcct gaatacataa
     5041 accagtccgt tcccaaaagg cccgctggct ctgtgcagaa tcctgtctat cacaatcagc
     5101 ctctgaaccc cgcgcccagc agagacccac actaccagga cccccacagc actgcagtgg
     5161 gcaaccccga gtatctcaac actgtccagc ccacctgtgt caacagcaca ttcgacagcc
     5221 ctgcccactg ggcccagaaa ggcagccacc aaattagcct ggacaaccct gactaccagc
     5281 aggacttctt tcccaaggaa gccaagccaa atggcatctt taagggctcc acagctgaaa
     5341 atgcagaata cctaagggtc gcgccacaaa gcagtgaatt tattggagca tagaacccag
     5401 ctttcttgta caaagtggtt gatatccagc acagtggcgg ccgctcgagt ctagagggcc
     5461 cgcggttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct acgcgtaccg
     5521 gttagtaatg agatccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     5581 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     5641 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     5701 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     5761 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     5821 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     5881 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     5941 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     6001 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     6061 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac acatatggtg
     6121 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     6181 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     6241 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     6301 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     6361 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     6421 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     6481 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     6541 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     6601 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     6661 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     6721 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     6781 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaatcgat
     6841 agatcctaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     6901 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     6961 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     7021 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     7081 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     7141 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     7201 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     7261 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     7321 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     7381 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctga
     7441 gatcctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     7501 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     7561 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     7621 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     7681 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     7741 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     7801 tcagagagtg agaggcccgg gttaattaag gaaagggcta gatcattctt gaagacgaaa
     7861 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     7921 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     7981 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     8041 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     8101 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     8161 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     8221 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     8281 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     8341 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     8401 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     8461 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     8521 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     8581 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     8641 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     8701 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     8761 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     8821 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     8881 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     8941 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     9001 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     9061 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     9121 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     9181 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     9241 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     9301 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     9361 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     9421 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     9481 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     9541 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     9601 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     9661 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     9721 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     9781 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     9841 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     9901 ttaatgcagc aagctcatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     9961 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
    10021 aaaaagctcc ccgtggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
    10081 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
    10141 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacat
    10201 gattacgaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
    10261 actcatcaat gtatcttatc atgtctggat caactggata actcaagcta accaaaatca
    10321 tcccaaactt cccaccccat accctattac cactgccaat tacctgtggt ttcatttact
    10381 ctaaacctgt gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg
    10441 tactacaaac ttagtagttg gaagggctaa ttcactccca aagaagacaa gatatccttg
    10501 atctgtggat ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc
    10561 caggggtcag atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag
    10621 ataaggtaga agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc
    10681 atgggatgga tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat
    10741 ttcatcacgt ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt
    10801 gctacaaggg actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg
    10861 agtggcgagc cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc
    10921 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
    10981 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
    11041 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
    11101 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
    11161 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
    11221 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
    11281 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
    11341 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
    11401 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
    11461 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
    11521 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
    11581 gtaagaccac cgcacagcaa gcggccggcc gctgatcttc agacctggag gaggagatat
    11641 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
    11701 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
    11761 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
    11821 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
    11881 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
    11941 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
    12001 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
    12061 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
    12121 taacaattac acaagcttaa tacac
//
