LOCUS       sequence-250708-       12857 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of PTCH1 G1212D.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12857)
  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 12857)
  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-G1212D
FEATURES             Location/Qualifiers
     source          1..12857
                     /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(7817..7841)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        7849..7867
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7892..7911)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7893..7911)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(7917..7937)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             7991..8032
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    8053..8620
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    8070..8622
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8214..8231)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8441..8460
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8634..9233
                     /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(8634..8653)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9130..9150
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9247..9835
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9300..9320)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9718..9729)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9910..10143
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10262..10280)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10348..10452
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10453..10521,10522..11313)
                     /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(10671..10690)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11484..12072
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11973..11992
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12226..12243
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12483..12504
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12519..12524,12525..12542,12543..12549)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12557..12573
                     /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     12562..12584
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12581..12597
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12581..12597
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12647..12666
                     /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 cgtggtccgc
     7081 ttcgccatgc cgcccgacca cacgcacagc gggtctgatt cctccgactc ggagtatagt
     7141 tcccagacga cagtgtcagg cctcagcgag gagcttcggc actacgaggc ccagcagggc
     7201 gcgggaggcc ctgcccacca agtgatcgtg gaagccacag aaaaccccgt cttcgcccac
     7261 tccactgtgg tccatcccga atccaggcat cacccaccct cgaacccgag acagcagccc
     7321 cacctggact cagggtccct gcctcccgga cggcaaggcc agcagccccg cagggacccc
     7381 cccagagaag gcttgtggcc acccccctac agaccgcgca gagacgcttt tgaaatttct
     7441 actgaagggc attctggccc tagcaatagg gcccgctggg gccctcgcgg ggcccgttct
     7501 cacaaccctc ggaacccagc gtccactgcc atgggcagct ccgtgcccgg ctactgccag
     7561 cccatcacca ctgtgacggc ttctgcctcc gtgactgtcg ccgtgcaccc gccgcctgtc
     7621 cctgggcctg ggcggaaccc ccgaggggga ctctgcccag gctaccctga gactgaccac
     7681 ggcctgtttg aggaccccca cgtgcctttc cacgtccggt gtgagaggag ggattcgaag
     7741 gtggaagtca ttgagctgca ggacgtggaa tgcgaggaga ggccccgggg aagcagctcc
     7801 aactcaggcc tcatgggccc agctttcttg tacaaagtgg tgatatccaa ttaaccctca
     7861 ctaaagggtt atgatatagt gagttgttat tccctatagt gagtcgtatt accacccaac
     7921 ttttctatac aaagtggttg atatccagca cagtggcggc cgctcgagtc tagagggccc
     7981 gcggttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg
     8041 ttagtaatga gatccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg
     8101 ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag
     8161 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc
     8221 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg
     8281 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag
     8341 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca
     8401 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt
     8461 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc
     8521 tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa
     8581 ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca catatgaccg
     8641 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     8701 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     8761 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     8821 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     8881 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     8941 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     9001 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     9061 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     9121 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     9181 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc taaatcgata
     9241 gatcctaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     9301 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     9361 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     9421 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     9481 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     9541 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     9601 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     9661 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     9721 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     9781 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgag
     9841 atcctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     9901 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
     9961 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
    10021 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
    10081 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
    10141 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
    10201 cagagagtga gaggcccggg ttaattaagg aaagggctag atcattcttg aagacgaaag
    10261 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
    10321 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
    10381 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
    10441 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
    10501 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
    10561 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
    10621 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
    10681 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
    10741 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
    10801 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
    10861 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
    10921 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
    10981 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
    11041 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
    11101 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
    11161 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
    11221 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
    11281 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
    11341 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
    11401 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
    11461 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
    11521 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
    11581 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
    11641 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
    11701 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
    11761 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
    11821 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
    11881 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
    11941 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
    12001 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
    12061 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
    12121 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
    12181 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
    12241 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
    12301 taatgcagca agctcatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
    12361 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
    12421 aaaagctccc cgtggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
    12481 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
    12541 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgacatg
    12601 attacgaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
    12661 ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat
    12721 cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc
    12781 taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt
    12841 actacaaact tagtagt
//
