LOCUS       sequence-250454-       12094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of EGFR R521K.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12094)
  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 12094)
  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-R521K
FEATURES             Location/Qualifiers
     source          1..12094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2223..3401
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2454..3392
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3349..3369
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    3421..3445
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(7054..7078)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        7086..7104
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7129..7148)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7130..7148)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(7154..7174)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             7228..7269
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    7290..7857
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    7307..7859
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7451..7468)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7678..7697
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7871..8470
                     /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(7871..7890)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8367..8387
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8484..9072
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8537..8557)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8955..8966)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9147..9380
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9499..9517)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9585..9689
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9690..9758,9759..10550)
                     /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"
     primer_bind     complement(9908..9927)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10721..11309
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11210..11229
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11463..11480
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11720..11741
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11756..11761,11762..11779,11780..11786)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11794..11810
                     /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     11799..11821
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11818..11834
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11818..11834
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11884..11903
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcgt
     2221 gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     2281 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     2341 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     2401 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
     2461 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     2521 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
     2581 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
     2641 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
     2701 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
     2761 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
     2821 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
     2881 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
     2941 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     3001 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     3061 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     3121 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     3181 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     3241 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     3301 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     3361 cagtggttca aagttttttt cttccatttc aggtgtcgtg aagcggccct gcagatatca
     3421 acaagtttgt acaaaaaagc aggcaccatg cgaccctccg ggacggccgg ggcagcgctc
     3481 ctggcgctgc tggctgcgct ctgcccggcg agtcgggctc tggaggaaaa gaaagtttgc
     3541 caaggcacga gtaacaagct cacgcagttg ggcacttttg aagatcattt tctcagcctc
     3601 cagaggatgt tcaataactg tgaggtggtc cttgggaatt tggaaattac ctatgtgcag
     3661 aggaattatg atctttcctt cttaaagacc atccaggagg tggctggtta tgtcctcatt
     3721 gccctcaaca cagtggagcg aattcctttg gaaaacctgc agatcatcag aggaaatatg
     3781 tactacgaaa attcctatgc cttagcagtc ttatctaact atgatgcaaa taaaaccgga
     3841 ctgaaggagc tgcccatgag aaatttacag gaaatcctgc atggcgccgt gcggttcagc
     3901 aacaaccctg ccctgtgcaa cgtggagagc atccagtggc gggacatagt cagcagtgac
     3961 tttctcagca acatgtcgat ggacttccag aaccacctgg gcagctgcca aaagtgtgat
     4021 ccaagctgtc ccaatgggag ctgctggggt gcaggagagg agaactgcca gaaactgacc
     4081 aaaatcatct gtgcccagca gtgctccggg cgctgccgtg gcaagtcccc cagtgactgc
     4141 tgccacaacc agtgtgctgc aggctgcaca ggcccccggg agagcgactg cctggtctgc
     4201 cgcaaattcc gagacgaagc cacgtgcaag gacacctgcc ccccactcat gctctacaac
     4261 cccaccacgt accagatgga tgtgaacccc gagggcaaat acagctttgg tgccacctgc
     4321 gtgaagaagt gtccccgtaa ttatgtggtg acagatcacg gctcgtgcgt ccgagcctgt
     4381 ggggccgaca gctatgagat ggaggaagac ggcgtccgca agtgtaagaa gtgcgaaggg
     4441 ccttgccgca aagtgtgtaa cggaataggt attggtgaat ttaaagactc actctccata
     4501 aatgctacga atattaaaca cttcaaaaac tgcacctcca tcagtggcga tctccacatc
     4561 ctgccggtgg catttagggg tgactccttc acacatactc ctcctctgga tccacaggaa
     4621 ctggatattc tgaaaaccgt aaaggaaatc acagggtttt tgctgattca ggcttggcct
     4681 gaaaacagga cggacctcca tgcctttgag aacctagaaa tcatacgcgg caggaccaag
     4741 caacatggtc agttttctct tgcagtcgtc agcctgaaca taacatcctt gggattacgc
     4801 tccctcaagg agataagtga tggagatgtg ataatttcag gaaacaaaaa tttgtgctat
     4861 gcaaatacaa taaactggaa aaaactgttt gggacctccg gtcagaaaac caaaattata
     4921 agcaacagag gtgaaaacag ctgcaaggcc acaggccagg tctgccatgc cttgtgctcc
     4981 cccgagggct gctggggccc ggagcccaag gactgcgtct cttgccggaa tgtcagccga
     5041 ggcagggaat gcgtggacaa gtgcaacctt ctggagggtg agccaaggga gtttgtggag
     5101 aactctgagt gcatacagtg ccacccagag tgcctgcctc aggccatgaa catcacctgc
     5161 acaggacggg gaccagacaa ctgtatccag tgtgcccact acattgacgg cccccactgc
     5221 gtcaagacct gcccggcagg agtcatggga gaaaacaaca ccctggtctg gaagtacgca
     5281 gacgccggcc atgtgtgcca cctgtgccat ccaaactgca cctacggatg cactgggcca
     5341 ggtcttgaag gctgtccaac gaatgggcct aagatcccgt ccatcgccac tgggatggtg
     5401 ggggccctcc tcttgctgct ggtggtggcc ctggggatcg gcctcttcat gcgaaggcgc
     5461 cacatcgttc ggaagcgcac gctgcggagg ctgctgcagg agagggagct tgtggagcct
     5521 cttacaccca gtggagaagc tcccaaccaa gctctcttga ggatcttgaa ggaaactgaa
     5581 ttcaaaaaga tcaaagtgct gggctccggt gcgttcggca cggtgtataa gggactctgg
     5641 atcccagaag gtgagaaagt taaaattccc gtcgctatca aggaattaag agaagcaaca
     5701 tctccgaaag ccaacaagga aatcctcgat gaagcctacg tgatggccag cgtggacaac
     5761 ccccacgtgt gccgcctgct gggcatctgc ctcacctcca ccgtgcagct catcacgcag
     5821 ctcatgccct tcggctgcct cctggactat gtccgggaac acaaagacaa tattggctcc
     5881 cagtacctgc tcaactggtg tgtgcagatc gcaaagggca tgaactactt ggaggaccgt
     5941 cgcttggtgc accgcgacct ggcagccagg aacgtactgg tgaaaacacc gcagcatgtc
     6001 aagatcacag attttgggct ggccaaactg ctgggtgcgg aagagaaaga ataccatgca
     6061 gaaggaggca aagtgcctat caagtggatg gcattggaat caattttaca cagaatctat
     6121 acccaccaga gtgatgtctg gagctacggg gtgaccgttt gggagttgat gacctttgga
     6181 tccaagccat atgacggaat ccctgccagc gagatctcct ccatcctgga gaaaggagaa
     6241 cgcctccctc agccacccat atgtaccatc gatgtctaca tgatcatggt caagtgctgg
     6301 atgatagacg cagatagtcg cccaaagttc cgtgagttga tcatcgaatt ctccaaaatg
     6361 gcccgagacc cccagcgcta ccttgtcatt cagggggatg aaagaatgca tttgccaagt
     6421 cctacagact ccaacttcta ccgtgccctg atggatgaag aagacatgga cgacgtggtg
     6481 gatgccgacg agtacctcat cccacagcag ggcttcttca gcagcccctc cacgtcacgg
     6541 actcccctcc tgagctctct gagtgcaacc agcaacaatt ccaccgtggc ttgcattgat
     6601 agaaatgggc tgcaaagctg tcccatcaag gaagacagct tcttgcagcg atacagctca
     6661 gaccccacag gcgccttgac tgaggacagc atagacgaca ccttcctccc agtgcctgaa
     6721 tacataaacc agtccgttcc caaaaggccc gctggctctg tgcagaatcc tgtctatcac
     6781 aatcagcctc tgaaccccgc gcccagcaga gacccacact accaggaccc ccacagcact
     6841 gcagtgggca accccgagta tctcaacact gtccagccca cctgtgtcaa cagcacattc
     6901 gacagccctg cccactgggc ccagaaaggc agccaccaaa ttagcctgga caaccctgac
     6961 taccagcagg acttctttcc caaggaagcc aagccaaatg gcatctttaa gggctccaca
     7021 gctgaaaatg cagaatacct aagggtcgca tggtgcctgc tttcttgtac aaagtggtga
     7081 tatccaatta accctcacta aaggaagatg atagttaagt gtaagttacc ctatagtgag
     7141 tcgtattacc acccaacttt tctatacaaa gtggttgata tccagcacag tggcggccgc
     7201 tcgagtctag agggcccgcg gttcgaaggt aagcctatcc ctaaccctct cctcggtctc
     7261 gattctacgc gtaccggtta gtaatgagat ccctnccccc cccctaacgt tactggccga
     7321 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     7381 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     7441 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     7501 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     7561 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     7621 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     7681 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     7741 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
     7801 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat
     7861 ggccacacat atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     7921 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     7981 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     8041 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     8101 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     8161 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     8221 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     8281 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     8341 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     8401 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     8461 cggtgcctaa atcgatagat cctaatcaac ctctggatta caaaatttgt gaaagattga
     8521 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     8581 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     8641 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     8701 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     8761 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     8821 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     8881 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     8941 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     9001 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     9061 ccgcctcccc gcctgagatc ctttaagacc aatgacttac aaggcagctg tagatcttag
     9121 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     9181 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     9241 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     9301 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     9361 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     9421 tgcaaagaaa tgaatatcag agagtgagag gcccgggtta attaaggaaa gggctagatc
     9481 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     9541 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     9601 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     9661 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     9721 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     9781 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     9841 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     9901 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     9961 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
    10021 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
    10081 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
    10141 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
    10201 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
    10261 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
    10321 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
    10381 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
    10441 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
    10501 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
    10561 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
    10621 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
    10681 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
    10741 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
    10801 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
    10861 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
    10921 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
    10981 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
    11041 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
    11101 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
    11161 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
    11221 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
    11281 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
    11341 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
    11401 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
    11461 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
    11521 cgcgttggcc gattcattaa tgcagcaagc tcatggctga ctaatttttt ttatttatgc
    11581 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
    11641 aggcctaggc ttttgcaaaa agctccccgt ggcacgacag gtttcccgac tggaaagcgg
    11701 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
    11761 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
    11821 gaaacagcta tgacatgatt acgaatttca caaataaagc atttttttca ctgcattcta
    11881 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaac tggataactc
    11941 aagctaacca aaatcatccc aaacttccca ccccataccc tattaccact gccaattacc
    12001 tgtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt
    12061 gtatttgtta aatatgtact acaaacttag tagt
//
