LOCUS       sequence-250563-       10790 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Lentiviral expression of BRAF E586K.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10790)
  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 10790)
  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-BRAF-E586K
FEATURES             Location/Qualifiers
     source          1..10790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1199..1223
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1224..1233
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3528..3552)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        3560..3578
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3603..3622)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3604..3622)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(3628..3648)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             3702..3743
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3764..4331
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3781..4333
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3925..3942)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4152..4171
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4345..4944
                     /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(4345..4364)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4841..4861
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4958..5546
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5011..5031)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5429..5440)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5621..5854
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5973..5991)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6059..6163
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6164..6232,6233..7024)
                     /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(6382..6401)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7195..7783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7684..7703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7937..7954
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8194..8215
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8230..8235,8236..8253,8254..8260)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8268..8284
                     /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     8273..8295
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8292..8308
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8292..8308
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8358..8377
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             8569..9202
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    9249..9374
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9871..10104
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10289..10333
                     /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    10626..10743
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgaa gcggccctgc agatatcaac
     1201 aagtttgtac aaaaaagcag gcaccatggc ggcgctgagc ggtggcggtg gtggcggcgc
     1261 ggagccgggc caggctctgt tcaacgggga catggagccc gaggccggcg ccggcgccgg
     1321 cgccgcggcc tcttcggctg cggaccctgc cattccggag gaggtgtgga atatcaaaca
     1381 aatgattaag ttgacacagg aacatataga ggccctattg gacaaatttg gtggggagca
     1441 taatccacca tcaatatatc tggaggccta tgaagaatac accagcaagc tagatgcact
     1501 ccaacaaaga gaacaacagt tattggaatc tctggggaac ggaactgatt tttctgtttc
     1561 tagctctgca tcaatggata ccgttacatc ttcttcctct tctagccttt cagtgctacc
     1621 ttcatctctt tcagtttttc aaaatcccac agatgtggca cggagcaacc ccaagtcacc
     1681 acaaaaacct atcgttagag tcttcctgcc caacaaacag aggacagtgg tacctgcaag
     1741 gtgtggagtt acagtccgag acagtctaaa gaaagcactg atgatgagag gtctaatccc
     1801 agagtgctgt gctgtttaca gaattcagga tggagagaag aaaccaattg gttgggacac
     1861 tgatatttcc tggcttactg gagaagaatt gcatgtggaa gtgttggaga atgttccact
     1921 tacaacacac aactttgtac gaaaaacgtt tttcacctta gcattttgtg acttttgtcg
     1981 aaagctgctt ttccagggtt tccgctgtca aacatgtggt tataaatttc accagcgttg
     2041 tagtacagaa gttccactga tgtgtgttaa ttatgaccaa cttgatttgc tgtttgtctc
     2101 caagttcttt gaacaccacc caataccaca ggaagaggcg tccttagcag agactgccct
     2161 aacatctgga tcatcccctt ccgcacccgc ctcggactct attgggcccc aaattctcac
     2221 cagtccgtct ccttcaaaat ccattccaat tccacagccc ttccgaccag cagatgaaga
     2281 tcatcgaaat caatttgggc aacgagaccg atcctcatca gctcccaatg tgcatataaa
     2341 cacaatagaa cctgtcaata ttgatgactt gattagagac caaggatttc gtggtgatgg
     2401 aggatcaacc acaggtttgt ctgctacccc ccctgcctca ttacctggct cactaactaa
     2461 cgtgaaagcc ttacagaaat ctccaggacc tcagcgagaa aggaagtcat cttcatcctc
     2521 agaagacagg aatcgaatga aaacacttgg tagacgggac tcgagtgatg attgggagat
     2581 tcctgatggg cagattacag tgggacaaag aattggatct ggatcatttg gaacagtcta
     2641 caagggaaag tggcatggtg atgtggcagt gaaaatgttg aatgtgacag cacctacacc
     2701 tcagcagtta caagccttca aaaatgaagt aggagtactc aggaaaacac gacatgtgaa
     2761 tatcctactc ttcatgggct attccacaaa gccacaactg gctattgtta cccagtggtg
     2821 tgagggctcc agcttgtatc accatctcca tatcattgag accaaatttg agatgatcaa
     2881 acttatagat attgcacgac agactgcaca gggcatggat tacttacacg ccaagtcaat
     2941 catccacaga gacctcaaga gtaataatat atttcttcat aaagacctca cagtaaaaat
     3001 aggtgatttt ggtctagcta cagtgaaatc tcgatggagt gggtcccatc agtttgaaca
     3061 gttgtctgga tccattttgt ggatggcacc agaagtcatc agaatgcaag ataaaaatcc
     3121 atacagcttt cagtcagatg tatatgcatt tggaattgtt ctgtatgaat tgatgactgg
     3181 acagttacct tattcaaaca tcaacaacag ggaccagata atttttatgg tgggacgagg
     3241 atacctgtct ccagatctca gtaaggtacg gagtaactgt ccaaaagcca tgaagagatt
     3301 aatggcagag tgcctcaaaa agaaaagaga tgagagacca ctctttcccc aaattctcgc
     3361 ctctattgag ctgctggccc gctcattgcc aaaaattcac cgcagtgcat cagaaccctc
     3421 cttgaatcgg gctggtttcc aaacagagga ttttagtcta tatgcttgtg cttctccaaa
     3481 aacacccatc caggcagggg gatatggtgc gtttcctgtc cactagaacc cagctttctt
     3541 gtacaaagtg gtgatatcca attaaccctc actaaaggtt taagtgagtt atagaagtag
     3601 taccctatag tgagtcgtat taccacccaa cttttctata caaagtggtt gatatccagc
     3661 acagtggcgg ccgctcgagt ctagagggcc cgcggttcga aggtaagcct atccctaacc
     3721 ctctcctcgg tctcgattct acgcgtaccg gttagtaatg agatccctcc ccccccccta
     3781 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     3841 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     3901 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     3961 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     4021 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     4081 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     4141 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     4201 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     4261 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     4321 cacgatgata atatggccac acatatgacc gagtacaagc ccacggtgcg cctcgccacc
     4381 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     4441 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     4501 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     4561 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     4621 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     4681 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     4741 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     4801 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     4861 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     4921 atgacccgca agcccggtgc ctaaatcgat agatcctaat caacctctgg attacaaaat
     4981 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5041 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5101 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5161 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5221 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     5281 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     5341 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     5401 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     5461 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     5521 gatctccctt tgggccgcct ccccgcctga gatcctttaa gaccaatgac ttacaaggca
     5581 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     5641 caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     5701 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     5761 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     5821 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
     5881 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggcccgg gttaattaag
     5941 gaaagggcta gatcattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     6001 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     6061 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6121 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     6181 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6241 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     6301 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     6361 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     6421 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     6481 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6541 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     6601 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6661 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     6721 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     6781 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6841 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6901 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6961 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     7021 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     7081 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     7141 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     7201 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7261 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7321 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     7381 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7441 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7501 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7561 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7621 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7681 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7741 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7801 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7861 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     7921 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     7981 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc aagctcatgg ctgactaatt
     8041 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     8101 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc ccgtggcacg acaggtttcc
     8161 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     8221 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     8281 acaatttcac acaggaaaca gctatgacat gattacgaat ttcacaaata aagcattttt
     8341 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     8401 caactggata actcaagcta accaaaatca tcccaaactt cccaccccat accctattac
     8461 cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg aattattttc
     8521 attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagttg gaagggctaa
     8581 ttcactccca aagaagacaa gatatccttg atctgtggat ctaccacaca caaggctact
     8641 tccctgatta gcagaactac acaccagggc caggggtcag atatccactg acctttggat
     8701 ggtgctacaa gctagtacca gttgagccag ataaggtaga agaggccaat aaaggagaga
     8761 acaccagctt gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtgt
     8821 tagagtggag gtttgacagc cgcctagcat ttcatcacgt ggcccgagag ctgcatccgg
     8881 agtacttcaa gaactgctga tatcgagctt gctacaaggg actttccgct ggggactttc
     8941 cagggaggcg tggcctgggc gggactgggg agtggcgagc cctcagatcc tgcatataag
     9001 cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     9061 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     9121 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     9181 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
     9241 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
     9301 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
     9361 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
     9421 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
     9481 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
     9541 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
     9601 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
     9661 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccggcc
     9721 gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa
     9781 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
     9841 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
     9901 agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt
     9961 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
    10021 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag
    10081 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
    10141 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca
    10201 cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt
    10261 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa
    10321 atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt
    10381 cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt
    10441 gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag
    10501 gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc
    10561 cattcgatta gtgaacggat ctcgacggta tcgccgaatt cacaaatggc agtattcatc
    10621 cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac
    10681 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat
    10741 tttcgggttt attacaggga cagcagagat ccagtttgga ctagtcgtga
//
