LOCUS       sequence-250714-       12856 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of PTCH1 S1203Rfs*52.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12856)
  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 12856)
  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-PTCH1-S1203Rfs*52
FEATURES             Location/Qualifiers
     source          1..12856
                     /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(7816..7840)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        7848..7866
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7891..7910)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7892..7910)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(7916..7936)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             7990..8031
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    8052..8619
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    8069..8621
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8213..8230)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8440..8459
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8633..9232
                     /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(8633..8652)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9129..9149
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9246..9834
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9299..9319)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9717..9728)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9909..10142
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10261..10279)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10347..10451
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10452..10520,10521..11312)
                     /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(10670..10689)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11483..12071
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11972..11991
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12225..12242
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12482..12503
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12518..12523,12524..12541,12542..12548)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12556..12572
                     /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     12561..12583
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12580..12596
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12580..12596
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12646..12665
                     /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 agaagggccg tcaaggccca ccatggcctc ggctggtaac
     3481 gccgccgagc cccaggaccg cggcggcggc ggcagcggct gtatcggtgc cccgggacgg
     3541 ccggctggag gcgggaggcg cagacggacg ggggggctgc gccgtgctgc cgcgccggac
     3601 cgggactatc tgcaccggcc cagctactgc gacgccgcct tcgctctgga gcagatttcc
     3661 aaggggaagg ctactggccg gaaagcgccg ctgtggctga gagcgaagtt tcagagactc
     3721 ttatttaaac tgggttgtta cattcaaaaa aactgcggca agttcttggt tgtgggcctc
     3781 ctcatatttg gggccttcgc ggtgggatta aaagcagcga acctcgagac caacgtggag
     3841 gagctgtggg tggaagttgg aggacgagta agtcgtgaat taaattatac tcgccagaag
     3901 attggagaag aggctatgtt taatcctcaa ctcatgatac agacccctaa agaagaaggt
     3961 gctaatgtcc tgaccacaga agcgctccta caacacctgg actcggcact ccaggccagc
     4021 cgtgtccatg tatacatgta caacaggcag tggaaattgg aacatttgtg ttacaaatca
     4081 ggagagctta tcacagaaac aggttacatg gatcagataa tagaatatct ttacccttgt
     4141 ttgattatta cacctttgga ctgcttctgg gaaggggcga aattacagtc tgggacagca
     4201 tacctcctag gtaaacctcc tttgcggtgg acaaacttcg accctttgga attcctggaa
     4261 gagttaaaga aaataaacta tcaagtggac agctgggagg aaatgctgaa taaggctgag
     4321 gttggtcatg gttacatgga ccgcccctgc ctcaatccgg ccgatccaga ctgccccgcc
     4381 acagccccca acaaaaattc aaccaaacct cttgatatgg cccttgtttt gaatggtgga
     4441 tgtcatggct tatccagaaa gtatatgcac tggcaggagg agttgattgt gggtggcaca
     4501 gtcaagaaca gcactggaaa actcgtcagc gcccatgccc tgcagaccat gttccagtta
     4561 atgactccca agcaaatgta cgagcacttc aaggggtacg agtatgtctc acacatcaac
     4621 tggaacgagg acaaagcggc agccatcctg gaggcctggc agaggacata tgtggaggtg
     4681 gttcatcaga gtgtcgcaca gaactccact caaaaggtgc tttccttcac caccacgacc
     4741 ctggacgaca tcctgaaatc cttctctgac gtcagtgtca tccgcgtggc cagcggctac
     4801 ttactcatgc tcgcctatgc ctgtctaacc atgctgcgct gggactgctc caagtcccag
     4861 ggtgccgtgg ggctggctgg cgtcctgctg gttgcactgt cagtggctgc aggactgggc
     4921 ctgtgctcat tgatcggaat ttcctttaac gctgcaacaa ctcaggtttt gccatttctc
     4981 gctcttggtg ttggtgtgga tgatgttttt cttctggccc acgccttcag tgaaacagga
     5041 cagaataaaa gaatcccttt tgaggacagg accggggagt gcctgaagcg cacaggagcc
     5101 agcgtggccc tcacgtccat cagcaatgtc acagccttct tcatggccgc gttaatccca
     5161 attcccgctc tgcgggcgtt ctccctccag gcagcggtag tagtggtgtt caattttgcc
     5221 atggttctgc tcatttttcc tgcaattctc agcatggatt tatatcgacg cgaggacagg
     5281 agactggata ttttctgctg ttttacaagc ccctgcgtca gcagagtgat tcaggttgaa
     5341 cctcaggcct acaccgacac acacgacaat acccgctaca gccccccacc tccctacagc
     5401 agccacagct ttgcccatga aacgcagatt accatgcagt ccactgtcca gctccgcacg
     5461 gagtacgacc cccacacgca cgtgtactac accaccgctg agccgcgctc cgagatctct
     5521 gtgcagcccg tcaccgtgac acaggacacc ctcagctgcc agagcccaga gagcaccagc
     5581 tccacaaggg acctgctctc ccagttctcc gactccagcc tccactgcct cgagcccccc
     5641 tgtacgaagt ggacactctc atcttttgct gagaagcact atgctccttt cctcttgaaa
     5701 ccaaaagcca aggtagtggt gatcttcctt tttctgggct tgctgggggt cagcctttat
     5761 ggcaccaccc gagtgagaga cgggctggac cttacggaca ttgtacctcg ggaaaccaga
     5821 gaatatgact ttattgctgc acaattcaaa tacttttctt tctacaacat gtatatagtc
     5881 acccagaaag cagactaccc gaatatccag cacttacttt acgacctaca caggagtttc
     5941 agtaacgtga agtatgtcat gttggaagaa aacaaacagc ttcccaaaat gtggctgcac
     6001 tacttcagag actggcttca gggacttcag gatgcatttg acagtgactg ggaaaccggg
     6061 aaaatcatgc caaacaatta caagaatgga tcagacgatg gagtccttgc ctacaaactc
     6121 ctggtgcaaa ccggcagccg cgataagccc atcgacatca gccagttgac taaacagcgt
     6181 ctggtggatg cagatggcat cattaatccc agcgctttct acatctacct gacggcttgg
     6241 gtcagcaacg accccgtcgc gtatgctgcc tcccaggcca acatccggcc acaccgacca
     6301 gaatgggtcc acgacaaagc cgactacatg cctgaaacaa ggctgagaat cccggcagca
     6361 gagcccatcg agtatgccca gttccctttc tacctcaacg gcttgcggga cacctcagac
     6421 tttgtggagg caattgaaaa agtaaggacc atctgcagca actatacgag cctggggctg
     6481 tccagttacc ccaacggcta ccccttcctc ttctgggagc agtacatcgg cctccgccac
     6541 tggctgctgc tgttcatcag cgtggtgttg gcctgcacat tcctcgtgtg cgctgtcttc
     6601 cttctgaacc cctggacggc cgggatcatt gtgatggtcc tggcgctgat gacggtcgag
     6661 ctgttcggca tgatgggcct catcggaatc aagctcagtg ccgtgcccgt ggtcatcctg
     6721 atcgcttctg ttggcatagg agtggagttc accgttcacg ttgctttggc ctttctgacg
     6781 gccatcggcg acaagaaccg cagggctgtg cttgccctgg agcacatgtt tgcacccgtc
     6841 ctggatggcg ccgtgtccac tctgctggga gtgctgatgc tggcgggatc tgagttcgac
     6901 ttcattgtca ggtatttctt tgctgtgctg gcgatcctca ccatcctcgg cgttctcaat
     6961 gggctggttt tgcttcccgt gcttttgtct ttctttggac catatcctga ggtgtctcca
     7021 gccaacggct tgaaccgcct gcccacaccc tcccctgagc caccccccag gtggtccgct
     7081 tcgccatgcc gcccggccac acgcacagcg ggtctgattc ctccgactcg gagtatagtt
     7141 cccagacgac agtgtcaggc ctcagcgagg agcttcggca ctacgaggcc cagcagggcg
     7201 cgggaggccc tgcccaccaa gtgatcgtgg aagccacaga aaaccccgtc ttcgcccact
     7261 ccactgtggt ccatcccgaa tccaggcatc acccaccctc gaacccgaga cagcagcccc
     7321 acctggactc agggtccctg cctcccggac ggcaaggcca gcagccccgc agggaccccc
     7381 ccagagaagg cttgtggcca cccccctaca gaccgcgcag agacgctttt gaaatttcta
     7441 ctgaagggca ttctggccct agcaataggg cccgctgggg ccctcgcggg gcccgttctc
     7501 acaaccctcg gaacccagcg tccactgcca tgggcagctc cgtgcccggc tactgccagc
     7561 ccatcaccac tgtgacggct tctgcctccg tgactgtcgc cgtgcacccg ccgcctgtcc
     7621 ctgggcctgg gcggaacccc cgagggggac tctgcccagg ctaccctgag actgaccacg
     7681 gcctgtttga ggacccccac gtgcctttcc acgtccggtg tgagaggagg gattcgaagg
     7741 tggaagtcat tgagctgcag gacgtggaat gcgaggagag gccccgggga agcagctcca
     7801 actcaggcct catgggccca gctttcttgt acaaagtggt gatatccaat taaccctcac
     7861 taaaggaata ttgaagatgt agtgagttat ccctatagtg agtcgtatta ccacccaact
     7921 tttctataca aagtggttga tatccagcac agtggcggcc gctcgagtct agagggcccg
     7981 cggttcgaag gtaagcctat ccctaaccct ctcctcggtc tcgattctac gcgtaccggt
     8041 tagtaatgag atccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     8101 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     8161 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     8221 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     8281 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     8341 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     8401 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     8461 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     8521 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     8581 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacac atatgaccga
     8641 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     8701 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     8761 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     8821 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     8881 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     8941 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     9001 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     9061 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     9121 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     9181 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct aaatcgatag
     9241 atcctaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     9301 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     9361 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     9421 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     9481 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     9541 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     9601 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     9661 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     9721 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     9781 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctgaga
     9841 tcctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     9901 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
     9961 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
    10021 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
    10081 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
    10141 cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc
    10201 agagagtgag aggcccgggt taattaagga aagggctaga tcattcttga agacgaaagg
    10261 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
    10321 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
    10381 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
    10441 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
    10501 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
    10561 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
    10621 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
    10681 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
    10741 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
    10801 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
    10861 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
    10921 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
    10981 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
    11041 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
    11101 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
    11161 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
    11221 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
    11281 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
    11341 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
    11401 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
    11461 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
    11521 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
    11581 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
    11641 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
    11701 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
    11761 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
    11821 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
    11881 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
    11941 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
    12001 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
    12061 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
    12121 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
    12181 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
    12241 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
    12301 aatgcagcaa gctcatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
    12361 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
    12421 aaagctcccc gtggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
    12481 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
    12541 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga
    12601 ttacgaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
    12661 tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc
    12721 ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt catttactct
    12781 aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta
    12841 ctacaaactt agtagt
//
