LOCUS       sequence-460590-       10552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet inducible expression of DmrB-Ripk3 with mCherry to induce Ripk3 
            necroptosis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10552)
  AUTHORS   Hanggi K, Li J, Gangadharan A, Liu X, Celias DP, Osunmakinde O, 
            Keske A, Davis J, Ahmad F, Giron A, Anadon CM, Gardner A, DeNardo 
            DG, Shaw TI, Beg AA, Yu X, Ruffell B
  TITLE     Interleukin-1alpha release during necrotic-like cell death generates
            myeloid-driven immunosuppression that restricts anti-tumor immunity.
  JOURNAL   Cancer Cell. 2024 Nov 13:S1535-6108(24)00402-1. doi: 
            10.1016/j.ccell.2024.10.014.
  PUBMED    39577420
REFERENCE   2  (bases 1 to 10552)
  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"; journalName: "Cancer 
            Cell. 2024 Nov 13:S1535-6108(24)00402-1. doi: 
            10.1016/j.ccell.2024.10.014."
COMMENT     Sequence Label: pLVX--DmrB-DmrB-Ripk3(deltaC)-2A-mCherry-puro
FEATURES             Location/Qualifiers
     source          1..10552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    680..805
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1302..1535
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1720..1764
                     /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    2027..2142
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        2205..2554
                     /label=TRE3GV promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, optimized for 
                     retroviral and lentiviral vectors"
     protein_bind    2213..2231
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2249..2267
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2285..2303
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2321..2339
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2357..2375
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2393..2411
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2429..2447
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2493..2517
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3926..4246
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             4262..4582
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             4601..4654
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4655..5362
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4800..4818)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5068..5087
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        5384..5883
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5419..5442)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5816..5835
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             5904..6503
                     /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(5904..5923)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6400..6420
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6520..7108
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6573..6593)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6991..7002)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7315..7948
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8076..8092)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8076..8092)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8089..8111)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8100..8116
                     /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)."
     promoter        complement(join(8124..8130,8131..8148,8149..8154))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8169..8190
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8307..8324)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8478..9066)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8558..8577)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9237..10028,10029..10097))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     9860..9879
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10098..10202)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    10250..10384
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10287..10306)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10341..10360
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             join(10552,1..633)
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 ggaagggcta attcactccc aaagaagaca agatatcctt gatctgtgga tctaccacac
       61 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact
      121 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa
      181 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga
      241 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga
      301 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc
      361 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc
      421 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag
      481 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
      541 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
      601 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
      661 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
      721 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
      781 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
      841 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
      901 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
      961 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     1021 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     1081 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     1141 agcggccggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
     1201 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1261 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1321 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1381 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1441 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1501 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1561 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1621 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1681 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1741 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1801 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1861 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1921 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1981 agagacagat ccattcgatt agtgaacgga tctcgacggt atcgccttta aaagaaaagg
     2041 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2101 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     2161 cagcagagat ccagtttatc gatgaggccc tttcgtcttc actcgagttt actccctatc
     2221 agtgatagag aacgtatgaa gagtttactc cctatcagtg atagagaacg tatgcagact
     2281 ttactcccta tcagtgatag agaacgtata aggagtttac tccctatcag tgatagagaa
     2341 cgtatgacca gtttactccc tatcagtgat agagaacgta tctacagttt actccctatc
     2401 agtgatagag aacgtatatc cagtttactc cctatcagtg atagagaacg tatgtcgagg
     2461 taggcgtgta cggtgggcgc ctataaaagc agagctcgtt tagtgaaccg tcagatcgcc
     2521 tggagcaatt ccacaacact tttgtcttat acttggatcc gggcccatgt cttctgtcaa
     2581 gttatggcct actggtgcgt cagcggttcc tctggtgagc cgtgaagaac tgaagaagct
     2641 ggagtttgtg ggcaaaggag ggttcggagt cgtgttccgg gcacaccaca gaacatggaa
     2701 ccatgatgta gcagtcaaga tcgtgaactc gaagaagata tcctgggagg tgaaggctat
     2761 ggttaatctt cgtaatgaga acgttctgct cctgctgggg gtcactgagg acctccagtg
     2821 ggacttcgtg tccgggcagg ctctggtgac aagattcatg gagaatggct ccctcgcagg
     2881 gctgctgcaa cccgagtgcc ctcggccctg gccactcctc tgtcgcctgc tgcaggaagt
     2941 ggtgctgggg atgtgctacc tacacagctt gaaccctccg ctcctgcacc gggacctcaa
     3001 gccctctaac attctgctgg atccagagct ccacgccaag ctagcagatt ttggcctgtc
     3061 cacgtttcag ggagggtccc agtcagggtc aggatcagga tcaggatcca gggactctgg
     3121 gggcacccta gcgtacttgg acccagagct gttatttaat gtcaacctga aggcttctaa
     3181 agcgagtgat gtctacagct ttgggatcct cgtgtgggca gtgctggctg gcagagaagc
     3241 tgagttggta gacaagactt cactaatccg ggaaacagtg tgtgacaggc agagtcgtcc
     3301 tccactgaca gagctgcctc caggtagccc tgagactcct ggcttggaaa aactgaagga
     3361 gttaatgatt cattgctggg gttcccagtc cgaaaacagg ccatccttcc aggactgcga
     3421 accaaaaacc aatgaagttt acaatctggt aaaggacaag gtagatgctg ctgtctccga
     3481 ggtaaagcat tatctgtctc agcacagaag cagcggcaga aacttgtctg ccagagagcc
     3541 aagccaaaga ggcacagaaa tggattgccc gagggaaacc atggtttcta aaatgctgga
     3601 ccgcctgcat ttggaggaac cctccggacc agttcctgga aaatgtcctg agaggcaagc
     3661 acaggacaca tcagttgggc ctgccacacc agcaaggaca tcttctgacc ccgtggctgg
     3721 cactcctcag attccacata ctttaccctt cagaggcaca acacctgggc cagtctttac
     3781 tgagactccc ggtcctcacc cccaaaggaa tcagggagat ggaagacacg gcactccttg
     3841 gtatccctgg accccaccga atccaatgac agggccaccg gctctcgtct tcaacaacgg
     3901 gcccagcggc cgcatggctt ctagaggagt gcaggtggag actatctccc caggagacgg
     3961 gcgcaccttc cccaagcgcg gccagacctg cgtggtgcac tacaccggga tgcttgaaga
     4021 tggaaagaaa gttgattcct cccgggacag aaacaagccc tttaagttta tgctaggcaa
     4081 gcaggaggtg atccgaggct gggaagaagg ggttgcccag atgagtgtgg gtcagagagc
     4141 caaactgact atatctccag attatgccta tggtgccact gggcacccag gcatcatccc
     4201 accacatgcc actctcgtct tcgatgtgga gcttctaaaa ctggaaacgc gtgcttctag
     4261 aggagtgcag gtggagacta tctccccagg agacgggcgc accttcccca agcgcggcca
     4321 gacctgcgtg gtgcactaca ccgggatgct tgaagatgga aagaaagttg attcctcccg
     4381 ggacagaaac aagcccttta agtttatgct aggcaagcag gaggtgatcc gaggctggga
     4441 agaaggggtt gcccagatga gtgtgggtca gagagccaaa ctgactatat ctccagatta
     4501 tgcctatggt gccactgggc acccaggcat catcccacca catgccactc tcgtcttcga
     4561 tgtggagctt ctaaaactgg aaacgcgtca tatggaattc gagggcagag gaagtcttct
     4621 aacatgcggt gacgtggagg agaatcccgg ccctatggtg agcaagggcg aggaggataa
     4681 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     4741 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     4801 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     4861 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     4921 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     4981 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     5041 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     5101 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     5161 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     5221 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     5281 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     5341 cggcatggac gagctgtaca aggaattctg actgactgat accgggtagg ggaggcgctt
     5401 ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca
     5461 agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc
     5521 tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc
     5581 ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg
     5641 cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag
     5701 cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg
     5761 ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc
     5821 tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt
     5881 tcgacctgca gcccaagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc
     5941 gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
     6001 cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct
     6061 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
     6121 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
     6181 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
     6241 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
     6301 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     6361 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
     6421 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
     6481 tgacccgcaa gcccggtgcc tgacgggcgc gtctggaaca atcaacctct ggattacaaa
     6541 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     6601 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     6661 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     6721 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     6781 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     6841 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     6901 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     6961 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     7021 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     7081 cggatctccc tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa
     7141 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag
     7201 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa
     7261 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag
     7321 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag
     7381 gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct
     7441 ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag
     7501 gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag
     7561 aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc
     7621 atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg
     7681 actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca
     7741 tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
     7801 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     7861 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     7921 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
     7981 ttataacttg caaagaaatg aatatcagag agtgagaggc cttgacattg ctagcgttta
     8041 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     8101 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     8161 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     8221 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     8281 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     8341 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     8401 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     8461 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     8521 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     8581 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     8641 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     8701 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     8761 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     8821 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     8881 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     8941 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     9001 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     9061 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     9121 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     9181 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     9241 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     9301 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     9361 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     9421 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     9481 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     9541 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     9601 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     9661 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     9721 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     9781 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     9841 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     9901 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     9961 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
    10021 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
    10081 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
    10141 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
    10201 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat
    10261 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
    10321 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
    10381 gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat
    10441 taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt
    10501 ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag tt
//
