LOCUS       sequence-460629-       10275 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet inducible expression of DmrB-Mlkl with mCherry to induce WT Mlkl
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10275)
  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 10275)
  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 MLKL(wt)--2A-mCherry-puro
FEATURES             Location/Qualifiers
     source          1..10275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     351..371
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    471..1059
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(524..544)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(942..953)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1266..1899
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(2027..2043)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2027..2043)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2040..2062)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2051..2067
                     /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(2075..2081,2082..2099,2100..2105))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2120..2141
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2258..2275)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2429..3017)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2509..2528)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3188..3979,3980..4048))
                     /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     3811..3830
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4049..4153)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    4201..4335
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4238..4257)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4292..4311
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             4503..5136
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5183..5308
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5805..6038
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6223..6267
                     /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    6530..6645
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        6708..7057
                     /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    6716..6734
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6752..6770
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6788..6806
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6824..6842
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6860..6878
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6896..6914
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6932..6950
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     6996..7020
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             8468..8521
                     /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             8522..9232
                     /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(8667..8685)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     8935..8954
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             9256..9576
                     /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"
     promoter        9610..10109
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(9645..9668)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     10042..10061
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             join(10130..10275,1..454)
                     /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(10130..10149)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
       61 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
      121 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
      181 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
      241 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
      301 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
      361 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
      421 cacctggtgc atgacccgca agcccggtgc ctgacgggcg cgtctggaac aatcaacctc
      481 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
      541 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
      601 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
      661 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
      721 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
      781 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
      841 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
      901 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
      961 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     1021 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattaat tctgcagtcg
     1081 agacctagaa aaacatggag caatcacaag tagcaataca gcagctacca atgctgattg
     1141 tgcctggcta gaagcacaag aggaggagga ggtgggtttt ccagtcacac ctcaggtacc
     1201 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaagag
     1261 gggactggaa gggctaattc actcccaacg aagacaagat atccttgatc tgtggatcta
     1321 ccacacacaa ggctacttcc ctgattagca gaactacaca ccagggccag gggtcagata
     1381 tccactgacc tttggatggt gctacaagct agtaccagtt gagccagata aggtagaaga
     1441 ggccaataaa ggagagaaca ccagcttgtt acaccctgtg agcctgcatg ggatggatga
     1501 cccggagaga gaagtgttag agtggaggtt tgacagccgc ctagcatttc atcacgtggc
     1561 ccgagagctg catccggagt acttcaagaa ctgctgatat cgagcttgct acaagggact
     1621 ttccgctggg gactttccag ggaggcgtgg cctgggcggg actggggagt ggcgagccct
     1681 cagatcctgc atataagcag ctgctttttg cctgtactgg gtctctctgg ttagaccaga
     1741 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     1801 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     1861 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta
     1921 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg ccttgacatt
     1981 gctagcgttt accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     2041 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     2101 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     2161 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2221 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2281 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2341 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2401 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2461 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2521 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2581 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2641 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2701 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2761 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2821 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2881 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2941 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3001 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3061 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3121 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3181 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3241 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3301 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3361 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3421 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3481 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3541 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3601 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3661 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3721 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3781 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3841 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3901 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3961 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4021 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4081 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4141 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     4201 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4261 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4321 tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc
     4381 ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac ctgtgattcc
     4441 tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta
     4501 gttggaaggg ctaattcact cccaaagaag acaagatatc cttgatctgt ggatctacca
     4561 cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc
     4621 actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc
     4681 caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc
     4741 ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg
     4801 agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc
     4861 cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag
     4921 atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct
     4981 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5041 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5101 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     5161 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     5221 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     5281 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     5341 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     5401 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     5461 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     5521 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     5581 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     5641 gcaagcggcc ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga
     5701 agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag
     5761 gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt
     5821 gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag
     5881 gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag
     5941 gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc
     6001 ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga
     6061 aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa
     6121 cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc
     6181 ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta
     6241 ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg
     6301 tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct
     6361 gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac
     6421 ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga
     6481 gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgcct ttaaaagaaa
     6541 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     6601 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     6661 ggacagcaga gatccagttt atcgatgagg ccctttcgtc ttcactcgag tttactccct
     6721 atcagtgata gagaacgtat gaagagttta ctccctatca gtgatagaga acgtatgcag
     6781 actttactcc ctatcagtga tagagaacgt ataaggagtt tactccctat cagtgataga
     6841 gaacgtatga ccagtttact ccctatcagt gatagagaac gtatctacag tttactccct
     6901 atcagtgata gagaacgtat atccagttta ctccctatca gtgatagaga acgtatgtcg
     6961 aggtaggcgt gtacggtggg cgcctataaa agcagagctc gtttagtgaa ccgtcagatc
     7021 gcctggagca attccacaac acttttgtct tatacttgga tccgggccca tggataaatt
     7081 gggacagatc atcaagttag gccagctcat ctatgaacag tgtgaaaaga tgaaatactg
     7141 ccggaaacaa tgccagcgtc taggaaaccg tgtgcacggc ctgctacagc ctctccagag
     7201 actccaggcc caaggaaaga agaacctgcc cgatgacatt actgctgccc tgggccgttt
     7261 tgatgaagtc ctgaaggagg ctaaccagca gatagaaaag ttcagcaaga agtcccatat
     7321 ttggaagttt gtgagtgtgg gcaatgataa gatcctcttc catgaagtga atgagaagct
     7381 gagagacgtc tgggaggagc tgttgctgct gcttcaggtt tatcattgga ataccgtttc
     7441 agatgtcagc cagccagcat cctggcagca ggaagatcga caggatgcag aggaagacgg
     7501 aaatgaaaat atgaaagtta tcctgatgca gttgcaaatt agcgtggaag aaatcaacaa
     7561 aaccctgaag caatgctcac taaaacccac acaggagatc ccacaagatc tccaaatcaa
     7621 ggagattcca aaggaacatc ttggacctcc gtggaccaaa ctgaagacaa gtaaaatgag
     7681 caccatttat agaggagagt atcacagatc tccagttacc atcaaagtat tcaacaaccc
     7741 ccaggccgaa agtgttggaa tagtgaggtt cactttcaat gacgagatca aaaccatgaa
     7801 gaaattcgat tctcccaaca tcttgcgtat atttgggatt tgcattgatc aaacagtgaa
     7861 gccccctgag ttctccattg tcatggagta ctgtgaactt ggaaccctga gggaactgct
     7921 ggatagagaa aaagacctca caatgagtgt gcgcagcctc ctagtcctga gggcagccag
     7981 aggcttatac aggctacacc attcggaaac actccacaga aacatcagca gctccagttt
     8041 cctcgtagcc ggaggctacc aagtaaagct tgcaggattt gagttaagca aaacacagaa
     8101 ttccatcagc cggacagcaa agagcactaa agcagagaga tccagttcaa cgatatatgt
     8161 ctcccctgag agactgaaaa atccattttg cctttatgac ataaaagctg aaatatatag
     8221 ctttggaatt gtactctggg aaattgccac tggaaagatc ccatttgaag gctgtgattc
     8281 taagaagatc cgggagctgg tggctgagga caagaagcag gaaccagtgg gtcaggattg
     8341 ccctgagttg ttgcgggaaa tcattaatga gtgtcgtgcc catgagccct cccaacggcc
     8401 ctctgtggac ggaatcttgg agagactgtc tgcggttgaa gaatccacgg acaagaaggt
     8461 ggggcccgag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc
     8521 tatggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     8581 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     8641 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     8701 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     8761 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     8821 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     8881 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     8941 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     9001 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     9061 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     9121 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     9181 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt gacccgcggc
     9241 cgcatggctt ctagaggagt gcaggtggag actatctccc caggagacgg gcgcaccttc
     9301 cccaagcgcg gccagacctg cgtggtgcac tacaccggga tgcttgaaga tggaaagaaa
     9361 gttgattcct cccgggacag aaacaagccc tttaagttta tgctaggcaa gcaggaggtg
     9421 atccgaggct gggaagaagg ggttgcccag atgagtgtgg gtcagagagc caaactgact
     9481 atatctccag attatgccta tggtgccact gggcacccag gcatcatccc accacatgcc
     9541 actctcgtct tcgatgtgga gcttctaaaa ctggaaacgc gtcatatgga attctgactg
     9601 actgataccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag
     9661 ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca
     9721 ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc
     9781 ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg
     9841 aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg
     9901 taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc
     9961 tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc
    10021 gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt
    10081 tctcctcttc ctcatctccg ggcctttcga cctgcagccc aagcttacca tgaccgagta
    10141 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
    10201 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
    10261 gcgggtcacc gagct
//
