LOCUS       sequence-250644-       12122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of EGFR P589L.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12122)
  AUTHORS   Ng PK, Li J, Jeong KJ, Shao S, Chen H, Tsang YH, Sengupta S, Wang Z,
            Bhavana VH, Tran R, Soewito S, Minussi DC, Moreno D, Kong K, 
            Dogruluk T, Lu H, Gao J, Tokheim C, Zhou DC, Johnson AM, Zeng J, Ip 
            CKM, Ju Z, Wester M, Yu S, Li Y, Vellano CP, Schultz N, Karchin R, 
            Ding L, Lu Y, Cheung LWT, Chen K, Shaw KR, Meric-Bernstam F, Scott 
            KL, Yi S, Sahni N, Liang H, Mills GB
  TITLE     Systematic Functional Annotation of Somatic Mutations in Cancer.
  JOURNAL   Cancer Cell. 2018 Mar 12;33(3):450-462.e10. doi: 
            10.1016/j.ccell.2018.01.021.
  PUBMED    29533785
REFERENCE   2  (bases 1 to 12122)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.ccell.2018.01"; journalName: "Cancer Cell"; date: 
            "2018-03-12- 12"; volume: "33"; issue: "3"; pages: "450-462.e10"
COMMENT     Sequence Label: pHAGE-EGFR-P589L
FEATURES             Location/Qualifiers
     source          1..12122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    192..309
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        357..1535
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          588..1526
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1483..1503
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1555..1579
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1582..1653,1654..3516,3517..3585,3586..5214)
                     /codon_start=1
                     /product="precursor of human epidermal growth factor 
                     receptor"
                     /label=EGFR
                     /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE
                     DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL
                     QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ
                     WRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR
                     CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP
                     EGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI
                     GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK
                     EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD
                     GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG
                     PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP
                     DNCIQCAHYIDGLHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG
                     CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP
                     SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(5216..5240)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        5248..5266
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5291..5310)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5292..5310)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(5316..5336)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             5390..5431
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    5452..6019
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5469..6021
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5613..5630)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5840..5859
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6033..6632
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6033..6052)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6529..6549
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6646..7234
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6699..6719)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7117..7128)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7309..7542
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7661..7679)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7747..7851
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7852..7920,7921..8712)
                     /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      8883..9471
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9372..9391
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9625..9642
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9882..9903
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9918..9923,9924..9941,9942..9948)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9956..9972
                     /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     9961..9983
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9980..9996
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9980..9996
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10046..10065
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             10257..10890
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10937..11062
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11559..11792
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11977..12021
                     /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 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
       61 cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga
      121 cagatccatt cgattagtga acggatctcg acggtatcgc cgaattcaca aatggcagta
      181 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
      241 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
      301 caaaattttc gggtttatta cagggacagc agagatccag tttggactag tcgtgaggct
      361 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
      421 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
      481 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
      541 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
      601 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
      661 ccacctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga
      721 gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg
      781 ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg
      841 ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag
      901 atagtcttgt aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg
      961 gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc
     1021 ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc
     1081 tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg
     1141 cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc
     1201 ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct
     1261 cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt
     1321 tctcgagctt ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt
     1381 ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct
     1441 ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg
     1501 ttcaaagttt ttttcttcca tttcaggtgt cgtgaagcgg ccctgcagat atcaacaagt
     1561 ttgtacaaaa aagcaggcac catgcgaccc tccgggacgg ccggggcagc gctcctggcg
     1621 ctgctggctg cgctctgccc ggcgagtcgg gctctggagg aaaagaaagt ttgccaaggc
     1681 acgagtaaca agctcacgca gttgggcact tttgaagatc attttctcag cctccagagg
     1741 atgttcaata actgtgaggt ggtccttggg aatttggaaa ttacctatgt gcagaggaat
     1801 tatgatcttt ccttcttaaa gaccatccag gaggtggctg gttatgtcct cattgccctc
     1861 aacacagtgg agcgaattcc tttggaaaac ctgcagatca tcagaggaaa tatgtactac
     1921 gaaaattcct atgccttagc agtcttatct aactatgatg caaataaaac cggactgaag
     1981 gagctgccca tgagaaattt acaggaaatc ctgcatggcg ccgtgcggtt cagcaacaac
     2041 cctgccctgt gcaacgtgga gagcatccag tggcgggaca tagtcagcag tgactttctc
     2101 agcaacatgt cgatggactt ccagaaccac ctgggcagct gccaaaagtg tgatccaagc
     2161 tgtcccaatg ggagctgctg gggtgcagga gaggagaact gccagaaact gaccaaaatc
     2221 atctgtgccc agcagtgctc cgggcgctgc cgtggcaagt cccccagtga ctgctgccac
     2281 aaccagtgtg ctgcaggctg cacaggcccc cgggagagcg actgcctggt ctgccgcaaa
     2341 ttccgagacg aagccacgtg caaggacacc tgccccccac tcatgctcta caaccccacc
     2401 acgtaccaga tggatgtgaa ccccgagggc aaatacagct ttggtgccac ctgcgtgaag
     2461 aagtgtcccc gtaattatgt ggtgacagat cacggctcgt gcgtccgagc ctgtggggcc
     2521 gacagctatg agatggagga agacggcgtc cgcaagtgta agaagtgcga agggccttgc
     2581 cgcaaagtgt gtaacggaat aggtattggt gaatttaaag actcactctc cataaatgct
     2641 acgaatatta aacacttcaa aaactgcacc tccatcagtg gcgatctcca catcctgccg
     2701 gtggcattta ggggtgactc cttcacacat actcctcctc tggatccaca ggaactggat
     2761 attctgaaaa ccgtaaagga aatcacaggg tttttgctga ttcaggcttg gcctgaaaac
     2821 aggacggacc tccatgcctt tgagaaccta gaaatcatac gcggcaggac caagcaacat
     2881 ggtcagtttt ctcttgcagt cgtcagcctg aacataacat ccttgggatt acgctccctc
     2941 aaggagataa gtgatggaga tgtgataatt tcaggaaaca aaaatttgtg ctatgcaaat
     3001 acaataaact ggaaaaaact gtttgggacc tccggtcaga aaaccaaaat tataagcaac
     3061 agaggtgaaa acagctgcaa ggccacaggc caggtctgcc atgccttgtg ctcccccgag
     3121 ggctgctggg gcccggagcc cagggactgc gtctcttgcc ggaatgtcag ccgaggcagg
     3181 gaatgcgtgg acaagtgcaa ccttctggag ggtgagccaa gggagtttgt ggagaactct
     3241 gagtgcatac agtgccaccc agagtgcctg cctcaggcca tgaacatcac ctgcacagga
     3301 cggggaccag acaactgtat ccagtgtgcc cactacattg acggcctcca ctgcgtcaag
     3361 acctgcccgg caggagtcat gggagaaaac aacaccctgg tctggaagta cgcagacgcc
     3421 ggccatgtgt gccacctgtg ccatccaaac tgcacctacg gatgcactgg gccaggtctt
     3481 gaaggctgtc caacgaatgg gcctaagatc ccgtccatcg ccactgggat ggtgggggcc
     3541 ctcctcttgc tgctggtggt ggccctgggg atcggcctct tcatgcgaag gcgccacatc
     3601 gttcggaagc gcacgctgcg gaggctgctg caggagaggg agcttgtgga gcctcttaca
     3661 cccagtggag aagctcccaa ccaagctctc ttgaggatct tgaaggaaac tgaattcaaa
     3721 aagatcaaag tgctgggctc cggtgcgttc ggcacggtgt ataagggact ctggatccca
     3781 gaaggtgaga aagttaaaat tcccgtcgct atcaaggaat taagagaagc aacatctccg
     3841 aaagccaaca aggaaatcct cgatgaagcc tacgtgatgg ccagcgtgga caacccccac
     3901 gtgtgccgcc tgctgggcat ctgcctcacc tccaccgtgc agctcatcac gcagctcatg
     3961 cccttcggct gcctcctgga ctatgtccgg gaacacaaag acaatattgg ctcccagtac
     4021 ctgctcaact ggtgtgtgca gatcgcaaag ggcatgaact acttggagga ccgtcgcttg
     4081 gtgcaccgcg acctggcagc caggaacgta ctggtgaaaa caccgcagca tgtcaagatc
     4141 acagattttg ggctggccaa actgctgggt gcggaagaga aagaatacca tgcagaagga
     4201 ggcaaagtgc ctatcaagtg gatggcattg gaatcaattt tacacagaat ctatacccac
     4261 cagagtgatg tctggagcta cggggtgacc gtttgggagt tgatgacctt tggatccaag
     4321 ccatatgacg gaatccctgc cagcgagatc tcctccatcc tggagaaagg agaacgcctc
     4381 cctcagccac ccatatgtac catcgatgtc tacatgatca tggtcaagtg ctggatgata
     4441 gacgcagata gtcgcccaaa gttccgtgag ttgatcatcg aattctccaa aatggcccga
     4501 gacccccagc gctaccttgt cattcagggg gatgaaagaa tgcatttgcc aagtcctaca
     4561 gactccaact tctaccgtgc cctgatggat gaagaagaca tggacgacgt ggtggatgcc
     4621 gacgagtacc tcatcccaca gcagggcttc ttcagcagcc cctccacgtc acggactccc
     4681 ctcctgagct ctctgagtgc aaccagcaac aattccaccg tggcttgcat tgatagaaat
     4741 gggctgcaaa gctgtcccat caaggaagac agcttcttgc agcgatacag ctcagacccc
     4801 acaggcgcct tgactgagga cagcatagac gacaccttcc tcccagtgcc tgaatacata
     4861 aaccagtccg ttcccaaaag gcccgctggc tctgtgcaga atcctgtcta tcacaatcag
     4921 cctctgaacc ccgcgcccag cagagaccca cactaccagg acccccacag cactgcagtg
     4981 ggcaaccccg agtatctcaa cactgtccag cccacctgtg tcaacagcac attcgacagc
     5041 cctgcccact gggcccagaa aggcagccac caaattagcc tggacaaccc tgactaccag
     5101 caggacttct ttcccaagga agccaagcca aatggcatct ttaagggctc cacagctgaa
     5161 aatgcagaat acctaagggt cgcgccacaa agcagtgaat ttattggagc atagaaccca
     5221 gctttcttgt acaaagtggt gatatccaat taaccctcac taaaggttta agtgagttat
     5281 agaagtagta ccctatagtg agtcgtatta ccacccaact tttctataca aagtggttga
     5341 tatccagcac agtggcggcc gctcgagtct agagggcccg cggttcgaag gtaagcctat
     5401 ccctaaccct ctcctcggtc tcgattctac gcgtaccggt tagtaatgag atccctcccc
     5461 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     5521 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     5581 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     5641 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc
     5701 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     5761 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     5821 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     5881 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
     5941 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     6001 ttgaaaaaca cgatgataat atggccacac atatgaccga gtacaagccc acggtgcgcc
     6061 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     6121 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     6181 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     6241 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     6301 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     6361 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     6421 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     6481 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     6541 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     6601 cctggtgcat gacccgcaag cccggtgcct aaatcgatag atcctaatca acctctggat
     6661 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     6721 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     6781 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     6841 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     6901 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     6961 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     7021 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     7081 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     7141 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     7201 acgagtcgga tctccctttg ggccgcctcc ccgcctgaga tcctttaaga ccaatgactt
     7261 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     7321 ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     7381 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     7441 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     7501 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     7561 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggcccgggt
     7621 taattaagga aagggctaga tcattcttga agacgaaagg gcctcgtgat acgcctattt
     7681 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     7741 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     7801 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     7861 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     7921 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     7981 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     8041 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     8101 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     8161 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     8221 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     8281 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     8341 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     8401 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     8461 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     8521 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     8581 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     8641 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     8701 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     8761 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     8821 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     8881 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     8941 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     9001 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     9061 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     9121 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     9181 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     9241 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     9301 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     9361 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     9421 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     9481 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     9541 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     9601 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     9661 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagcaa gctcatggct
     9721 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     9781 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccc gtggcacgac
     9841 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     9901 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     9961 agcggataac aatttcacac aggaaacagc tatgacatga ttacgaattt cacaaataaa
    10021 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
    10081 gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc caccccatac
    10141 cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa
    10201 ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagttgga
    10261 agggctaatt cactcccaaa gaagacaaga tatccttgat ctgtggatct accacacaca
    10321 aggctacttc cctgattagc agaactacac accagggcca ggggtcagat atccactgac
    10381 ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag aggccaataa
    10441 aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg acccggagag
    10501 agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg cccgagagct
    10561 gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac tttccgctgg
    10621 ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatcctg
    10681 catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct
    10741 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
    10801 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
    10861 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
    10921 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
    10981 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
    11041 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
    11101 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
    11161 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
    11221 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
    11281 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
    11341 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
    11401 ggccggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
    11461 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
    11521 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
    11581 ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga
    11641 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
    11701 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
    11761 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
    11821 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
    11881 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
    11941 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
    12001 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
    12061 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
    12121 tc
//
