LOCUS       sequence-250529-       12145 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Lentiviral expression of EGFR G588S.
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 Apr 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-G588S
FEATURES             Location/Qualifiers
     source          1..12145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    136..253
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        301..1479
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          532..1470
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1427..1447
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1499..1523
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1526..1597,1598..3460,3461..3529,3530..5158)
                     /codon_start=1
                     /product="precursor of human epidermal growth factor 
                     receptor"
                     /label=EGFR
                     /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE
                     DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL
                     QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ
                     WRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR
                     CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP
                     EGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI
                     GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK
                     EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD
                     GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG
                     PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP
                     DNCIQCAHYIDSPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG
                     CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP
                     SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(5160..5184)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5237..5278
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    5299..5866
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5316..5868
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5460..5477)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5687..5706
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5880..5882,5883..5885,5886..6599)
                     /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(5880..5882,5883..5885,5886..6596)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5925..5946)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6186..6205)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6533..6554
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6613..7201
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6666..6686)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7084..7095)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7276..7509
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7628..7646)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7714..7818
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7819..7887,7888..8679)
                     /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      8850..9438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9339..9358
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9592..9609
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9849..9870
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9885..9890,9891..9908,9909..9915)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9923..9939
                     /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     9928..9950
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9947..9963
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9947..9963
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10013..10032
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             10224..10857
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10904..11029
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11526..11759
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11944..11988
                     /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 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
       61 gagacagatc cattcgatta gtgaacggat ctcgacggta tcgccgaatt cacaaatggc
      121 agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag
      181 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa
      241 aattcaaaat tttcgggttt attacaggga cagcagagat ccagtttgga ctagtcgtga
      301 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
      361 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      421 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      481 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      541 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      601 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      661 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      721 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      781 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      841 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      901 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      961 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
     1021 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1081 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1141 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1201 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1261 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1321 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1381 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1441 gtggttcaaa gtttttttct tccatttcag gtgtcgtgaa gcggccctgc agatatcaac
     1501 aagtttgtac aaaaaagcag gcaccatgcg accctccggg acggccgggg cagcgctcct
     1561 ggcgctgctg gctgcgctct gcccggcgag tcgggctctg gaggaaaaga aagtttgcca
     1621 aggcacgagt aacaagctca cgcagttggg cacttttgaa gatcattttc tcagcctcca
     1681 gaggatgttc aataactgtg aggtggtcct tgggaatttg gaaattacct atgtgcagag
     1741 gaattatgat ctttccttct taaagaccat ccaggaggtg gctggttatg tcctcattgc
     1801 cctcaacaca gtggagcgaa ttcctttgga aaacctgcag atcatcagag gaaatatgta
     1861 ctacgaaaat tcctatgcct tagcagtctt atctaactat gatgcaaata aaaccggact
     1921 gaaggagctg cccatgagaa atttacagga aatcctgcat ggcgccgtgc ggttcagcaa
     1981 caaccctgcc ctgtgcaacg tggagagcat ccagtggcgg gacatagtca gcagtgactt
     2041 tctcagcaac atgtcgatgg acttccagaa ccacctgggc agctgccaaa agtgtgatcc
     2101 aagctgtccc aatgggagct gctggggtgc aggagaggag aactgccaga aactgaccaa
     2161 aatcatctgt gcccagcagt gctccgggcg ctgccgtggc aagtccccca gtgactgctg
     2221 ccacaaccag tgtgctgcag gctgcacagg cccccgggag agcgactgcc tggtctgccg
     2281 caaattccga gacgaagcca cgtgcaagga cacctgcccc ccactcatgc tctacaaccc
     2341 caccacgtac cagatggatg tgaaccccga gggcaaatac agctttggtg ccacctgcgt
     2401 gaagaagtgt ccccgtaatt atgtggtgac agatcacggc tcgtgcgtcc gagcctgtgg
     2461 ggccgacagc tatgagatgg aggaagacgg cgtccgcaag tgtaagaagt gcgaagggcc
     2521 ttgccgcaaa gtgtgtaacg gaataggtat tggtgaattt aaagactcac tctccataaa
     2581 tgctacgaat attaaacact tcaaaaactg cacctccatc agtggcgatc tccacatcct
     2641 gccggtggca tttaggggtg actccttcac acatactcct cctctggatc cacaggaact
     2701 ggatattctg aaaaccgtaa aggaaatcac agggtttttg ctgattcagg cttggcctga
     2761 aaacaggacg gacctccatg cctttgagaa cctagaaatc atacgcggca ggaccaagca
     2821 acatggtcag ttttctcttg cagtcgtcag cctgaacata acatccttgg gattacgctc
     2881 cctcaaggag ataagtgatg gagatgtgat aatttcagga aacaaaaatt tgtgctatgc
     2941 aaatacaata aactggaaaa aactgtttgg gacctccggt cagaaaacca aaattataag
     3001 caacagaggt gaaaacagct gcaaggccac aggccaggtc tgccatgcct tgtgctcccc
     3061 cgagggctgc tggggcccgg agcccaggga ctgcgtctct tgccggaatg tcagccgagg
     3121 cagggaatgc gtggacaagt gcaaccttct ggagggtgag ccaagggagt ttgtggagaa
     3181 ctctgagtgc atacagtgcc acccagagtg cctgcctcag gccatgaaca tcacctgcac
     3241 aggacgggga ccagacaact gtatccagtg tgcccactac attgacagcc cccactgcgt
     3301 caagacctgc ccggcaggag tcatgggaga aaacaacacc ctggtctgga agtacgcaga
     3361 cgccggccat gtgtgccacc tgtgccatcc aaactgcacc tacggatgca ctgggccagg
     3421 tcttgaaggc tgtccaacga atgggcctaa gatcccgtcc atcgccactg ggatggtggg
     3481 ggccctcctc ttgctgctgg tggtggccct ggggatcggc ctcttcatgc gaaggcgcca
     3541 catcgttcgg aagcgcacgc tgcggaggct gctgcaggag agggagcttg tggagcctct
     3601 tacacccagt ggagaagctc ccaaccaagc tctcttgagg atcttgaagg aaactgaatt
     3661 caaaaagatc aaagtgctgg gctccggtgc gttcggcacg gtgtataagg gactctggat
     3721 cccagaaggt gagaaagtta aaattcccgt cgctatcaag gaattaagag aagcaacatc
     3781 tccgaaagcc aacaaggaaa tcctcgatga agcctacgtg atggccagcg tggacaaccc
     3841 ccacgtgtgc cgcctgctgg gcatctgcct cacctccacc gtgcagctca tcacgcagct
     3901 catgcccttc ggctgcctcc tggactatgt ccgggaacac aaagacaata ttggctccca
     3961 gtacctgctc aactggtgtg tgcagatcgc aaagggcatg aactacttgg aggaccgtcg
     4021 cttggtgcac cgcgacctgg cagccaggaa cgtactggtg aaaacaccgc agcatgtcaa
     4081 gatcacagat tttgggctgg ccaaactgct gggtgcggaa gagaaagaat accatgcaga
     4141 aggaggcaaa gtgcctatca agtggatggc attggaatca attttacaca gaatctatac
     4201 ccaccagagt gatgtctgga gctacggggt gaccgtttgg gagttgatga cctttggatc
     4261 caagccatat gacggaatcc ctgccagcga gatctcctcc atcctggaga aaggagaacg
     4321 cctccctcag ccacccatat gtaccatcga tgtctacatg atcatggtca agtgctggat
     4381 gatagacgca gatagtcgcc caaagttccg tgagttgatc atcgaattct ccaaaatggc
     4441 ccgagacccc cagcgctacc ttgtcattca gggggatgaa agaatgcatt tgccaagtcc
     4501 tacagactcc aacttctacc gtgccctgat ggatgaagaa gacatggacg acgtggtgga
     4561 tgccgacgag tacctcatcc cacagcaggg cttcttcagc agcccctcca cgtcacggac
     4621 tcccctcctg agctctctga gtgcaaccag caacaattcc accgtggctt gcattgatag
     4681 aaatgggctg caaagctgtc ccatcaagga agacagcttc ttgcagcgat acagctcaga
     4741 ccccacaggc gccttgactg aggacagcat agacgacacc ttcctcccag tgcctgaata
     4801 cataaaccag tccgttccca aaaggcccgc tggctctgtg cagaatcctg tctatcacaa
     4861 tcagcctctg aaccccgcgc ccagcagaga cccacactac caggaccccc acagcactgc
     4921 agtgggcaac cccgagtatc tcaacactgt ccagcccacc tgtgtcaaca gcacattcga
     4981 cagccctgcc cactgggccc agaaaggcag ccaccaaatt agcctggaca accctgacta
     5041 ccagcaggac ttctttccca aggaagccaa gccaaatggc atctttaagg gctccacagc
     5101 tgaaaatgca gaatacctaa gggtcgcgcc acaaagcagt gaatttattg gagcatagaa
     5161 cccagctttc ttgtacaaag tggttgatat ccagcacagt ggcggccgct cgagtctaga
     5221 gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg
     5281 taccggttag taatgagatc cctccccccc ccctaacgtt actggccgaa gccgcttgga
     5341 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     5401 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     5461 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     5521 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     5581 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     5641 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     5701 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     5761 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     5821 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacacata
     5881 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     5941 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     6001 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     6061 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     6121 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     6181 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     6241 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     6301 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     6361 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     6421 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     6481 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     6541 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa
     6601 tcgatagatc ctaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     6661 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     6721 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     6781 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     6841 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     6901 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     6961 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     7021 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     7081 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     7141 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     7201 cctgagatcc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     7261 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg
     7321 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     7381 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     7441 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     7501 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     7561 gaatatcaga gagtgagagg cccgggttaa ttaaggaaag ggctagatca ttcttgaaga
     7621 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     7681 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     7741 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     7801 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     7861 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     7921 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     7981 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     8041 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     8101 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     8161 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     8221 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     8281 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     8341 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     8401 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     8461 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     8521 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     8581 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     8641 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     8701 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     8761 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     8821 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     8881 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     8941 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     9001 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     9061 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     9121 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     9181 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     9241 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     9301 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     9361 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     9421 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     9481 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     9541 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     9601 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     9661 attcattaat gcagcaagct catggctgac taattttttt tatttatgca gaggccgagg
     9721 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     9781 tttgcaaaaa gctccccgtg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     9841 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     9901 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     9961 gacatgatta cgaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
    10021 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa
    10081 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat
    10141 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa
    10201 atatgtacta caaacttagt agttggaagg gctaattcac tcccaaagaa gacaagatat
    10261 ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga actacacacc
    10321 agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga
    10381 gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag
    10441 cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct
    10501 agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg
    10561 agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac
    10621 tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt
    10681 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
    10741 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
    10801 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
    10861 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
    10921 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
    10981 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
    11041 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
    11101 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
    11161 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
    11221 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
    11281 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
    11341 caaaagtaag accaccgcac agcaagcggc cggccgctga tcttcagacc tggaggagga
    11401 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
    11461 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
    11521 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
    11581 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
    11641 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
    11701 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
    11761 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
    11821 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
    11881 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
    11941 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
    12001 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
    12061 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
    12121 ccattatcgt ttcagaccca cctcc
//
