LOCUS       sequence-460570-       10678 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 10678)
  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 10678)
  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-Ripk3-2A-mCherry-puro
FEATURES             Location/Qualifiers
     source          1..10678
                     /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             3916..3927
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             4052..4372
                     /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             4388..4708
                     /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             4727..4780
                     /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             4781..5488
                     /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(4926..4944)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5194..5213
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        5510..6009
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5545..5568)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5942..5961
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6030..6629
                     /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(6030..6049)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6526..6546
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6646..7234
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6699..6719)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7117..7128)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7441..8074
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8202..8218)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8202..8218)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8215..8237)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8226..8242
                     /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(8250..8256,8257..8274,8275..8280))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8295..8316
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8433..8450)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8604..9192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8684..8703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9363..10154,10155..10223))
                     /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     9986..10005
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10224..10328)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    10376..10510
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10413..10432)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10467..10486
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             join(10678,1..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 tcaacaactg
     3901 ttctgaagtg cagattggga actacaactc cttggtagca ccaccaagaa ctactgcctc
     3961 aagttcggcc aagtatgacc aagcacagtt cggcaggggt aggggctggc agcccttcca
     4021 caaggggccc agcggccgca tggcttctag aggagtgcag gtggagacta tctccccagg
     4081 agacgggcgc accttcccca agcgcggcca gacctgcgtg gtgcactaca ccgggatgct
     4141 tgaagatgga aagaaagttg attcctcccg ggacagaaac aagcccttta agtttatgct
     4201 aggcaagcag gaggtgatcc gaggctggga agaaggggtt gcccagatga gtgtgggtca
     4261 gagagccaaa ctgactatat ctccagatta tgcctatggt gccactgggc acccaggcat
     4321 catcccacca catgccactc tcgtcttcga tgtggagctt ctaaaactgg aaacgcgtgc
     4381 ttctagagga gtgcaggtgg agactatctc cccaggagac gggcgcacct tccccaagcg
     4441 cggccagacc tgcgtggtgc actacaccgg gatgcttgaa gatggaaaga aagttgattc
     4501 ctcccgggac agaaacaagc cctttaagtt tatgctaggc aagcaggagg tgatccgagg
     4561 ctgggaagaa ggggttgccc agatgagtgt gggtcagaga gccaaactga ctatatctcc
     4621 agattatgcc tatggtgcca ctgggcaccc aggcatcatc ccaccacatg ccactctcgt
     4681 cttcgatgtg gagcttctaa aactggaaac gcgtcatatg gaattcgagg gcagaggaag
     4741 tcttctaaca tgcggtgacg tggaggagaa tcccggccct atggtgagca agggcgagga
     4801 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
     4861 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     4921 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
     4981 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
     5041 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     5101 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
     5161 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
     5221 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     5281 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
     5341 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
     5401 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
     5461 caccggcggc atggacgagc tgtacaagga attctgactg actgataccg ggtaggggag
     5521 gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc
     5581 tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct
     5641 ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc
     5701 cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt
     5761 ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc
     5821 caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg
     5881 ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg
     5941 gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg
     6001 ggcctttcga cctgcagccc aagcttacca tgaccgagta caagcccacg gtgcgcctcg
     6061 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     6121 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     6181 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     6241 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     6301 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     6361 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     6421 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     6481 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     6541 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     6601 ggtgcatgac ccgcaagccc ggtgcctgac gggcgcgtct ggaacaatca acctctggat
     6661 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     6721 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     6781 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     6841 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     6901 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     6961 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     7021 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     7081 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     7141 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     7201 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc
     7261 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct
     7321 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa
     7381 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aagaggggac
     7441 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca
     7501 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
     7561 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
     7621 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
     7681 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
     7741 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
     7801 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
     7861 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
     7921 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     7981 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     8041 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
     8101 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattgctag
     8161 cgtttaccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     8221 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     8281 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     8341 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     8401 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     8461 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     8521 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     8581 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     8641 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     8701 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     8761 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     8821 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     8881 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     8941 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     9001 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     9061 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     9121 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     9181 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     9241 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     9301 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     9361 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     9421 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     9481 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     9541 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     9601 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     9661 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     9721 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     9781 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     9841 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     9901 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     9961 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
    10021 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
    10081 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
    10141 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
    10201 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
    10261 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
    10321 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt
    10381 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
    10441 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
    10501 tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata
    10561 ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga
    10621 attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagtt
//
