LOCUS       sequence-331188-       12974 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral plasmid with 3xFlag-pyrin (S208C/PAAND variant) under the
            control of a doxycycline-inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12974)
  AUTHORS   Chirita D, Jamilloux Y, Henry T, Magnotti F
  TITLE     Functional Assessment of Disease-Associated Pyrin Variants.
  JOURNAL   Methods Mol Biol. 2022;2523:179-195. doi: 
            10.1007/978-1-0716-2449-4_12.
  PUBMED    35759198
REFERENCE   2  (bases 1 to 12974)
  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: "Methods Mol
            Biol."; date: "2022"; volume: "2523:179-195. doi"; pages: " 
            10.1007/978-1-0716-2449-4_12"
COMMENT     Sequence Label: pInducer21-3xFlag-S208C-MEFV
FEATURES             Location/Qualifiers
     source          1..12974
                     /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"
     protein_bind    join(2230..2248,2249..2271,2272..2290,2291..2313,2314..2332,
                     2333..2355,2356..2374,2375..2397,2398..2416,2417..2439,
                     2440..2458,2459..2481,2482..2500)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2533..2571
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2568..2592
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2701..2725
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2768..2833
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(5203..5227)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5280..5306
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        5526..6704
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5757..6695
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     6652..6672
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    6774..6807
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             join(6814..7431,7432..7437,7438..7473,7474..7476,7477..7512,
                     7513..7515,7516..7551,7552..7560)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    7613..8186
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7780..7797)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8007..8026
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(8202..8204,8205..8207,8208..8921)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(8202..8204,8205..8207,8208..8918)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8247..8268)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8508..8527)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8855..8876
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     9057..9076
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    9364..9952
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9417..9437)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9835..9846)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10027..10260
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10379..10397)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10465..10569
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10570..10638,10639..11430)
                     /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(10788..10807)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11601..12189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12090..12109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12343..12360
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12600..12621
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12636..12641,12642..12659,12660..12666)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12674..12690
                     /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     12679..12701
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12698..12714
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12698..12714
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12764..12783
                     /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 gactaggatc
     2221 ctttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca
     2281 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa
     2341 agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
     2401 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
     2461 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagctcgg
     2521 tacccgggtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg
     2581 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
     2641 gaccgatcca gcctccgcgg ccccgaacta gtccagtgtg gtggaattct gcagatatca
     2701 acaagtttgt acaaaaaagc aggctttaaa ggaaccaatt cagtcgactg gatccggtac
     2761 caccatggac tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga
     2821 tgacgatgac aagcttgcgg ccgcggctaa gacccctagt gaccatctgc tgtccaccct
     2881 ggaggagctg gtgccctatg acttcgagaa gttcaagttc aagctgcaga acaccagtgt
     2941 gcagaaggag cactccagga tcccccggag ccagatccag agagccaggc cggtgaagat
     3001 ggccactctg ctggtcacct actatgggga agagtacgcc gtgcagctca ccctgcaggt
     3061 cctgcgggcc atcaaccagc gcctgctggc cgaggagctc cacagggcag ccattcagga
     3121 atattccaca caagaaaacg gcacagatga ttccgcagcg tccagctccc tgggggagaa
     3181 caagcccagg agcctgaaga ctccagacca ccccgagggg aacgagggga acggccctcg
     3241 gccgtacggg ggcggagctg ccagcctgcg gtgcagccag cccgaggccg ggagggggct
     3301 gtcgaggaag cccctgagca aacgcagaga gaaggcctcg gagggcctgg acgcgcaggg
     3361 caagcctcgg acccggagcc cggccctgcc gggcgggaga agccccggcc cctgcagggc
     3421 gctagagggg ggccaggccg aggtccggct gcgcagaaac gcctgctccg cggggaggct
     3481 gcaggggctg gcggggggcg ccccggggca gaaggagtgc aggcccttcg aagtgtacct
     3541 gccctcggga aagatgcgac ctagaagcct tgaggtcacc atttctacag gggagaaggc
     3601 gcccgcaaat ccagaaattc tcctgactct agaggaaaag acagctgcga atctggactc
     3661 ggcaacagaa ccccgggcaa ggcccactcc ggatggaggg gcatctgcgg acctgaagga
     3721 aggccctgga aatccagaac attcggtcac cggaaggcca ccagacacgg ctgcgagtcc
     3781 ccgctgccac gcccaggaag gagacccagt tgacggtacc tgtgtgcgtg attcctgcag
     3841 cttccccgag gcagtttctg ggcaccccca ggcctcaggc agccgctcac ctggctgccc
     3901 ccggtgccag gactcccatg aaaggaagag cccgggaagc ctaagccccc agcccctgcc
     3961 acagtgtaag cgccacctga agcaggtcca gctgctcttc tgtgaggatc acgatgagcc
     4021 catctgcctc atctgcagtc tgagtcagga gcaccaaggc caccgggtgc gccccattga
     4081 ggaggtcgcc ctggaacaca agaagaaaat tcagaagcag ctggagcatc tgaagaagct
     4141 gagaaaatca ggggaggagc agcgatccta tggggaggag aaggcagtga gctttctgaa
     4201 acaaactgaa gcgctgaagc agcgggtgca gaggaagctg gagcaggtgt actacttcct
     4261 ggagcagcaa gagcatttct ttgtggcctc actggaggac gtgggccaga tggttgggca
     4321 gatcaggaag gcatatgaca cccgcgtatc ccaggacatc gccctgctcg atgcgctgat
     4381 tggggaactg gaggccaagg agtgccagtc agaatgggaa cttctgcagg acattggaga
     4441 catcttgcac agggctaaga cagtgcctgt ccctgaaaag tggaccactc ctcaagagat
     4501 aaaacaaaag atccaactcc tccaccagaa gtcagagttt gtggagaaga gcacaaagta
     4561 cttctcagaa accctgcgtt cagaaatgga aatgttcaat gttccagagc tgattggcgc
     4621 tcaggcacat gctgttaatg tgattctgga tgcagaaacc gcttacccca acctcatctt
     4681 ctctgatgat ctgaagagtg ttagacttgg aaacaagtgg gagaggctgc ctgatggccc
     4741 gcaaagattt gacagctgta tcattgttct gggctctccg agtttcctct ctggccgccg
     4801 ttactgggag gtggaggttg gagacaagac agcatggatc ctgggagcct gcaagacatc
     4861 cataagcagg aaagggaaca tgactctgtc gccagagaat ggctactggg tggtgataat
     4921 gatgaaggaa aatgagtacc aggcgtccag cgttcccccg acccgcctgc taataaagga
     4981 gcctcccaag cgtgtgggca tcttcgtgga ctacagagtt ggaagcatct ccttttacaa
     5041 tgtgacagcc agatcccaca tctatacatt cgccagctgc tctttctctg ggccccttca
     5101 acctatcttc agccctggga cacgtgatgg agggaagaac acagctcctc tgactatctg
     5161 tccagtgggt ggtcaggggc ctgactgact cgagatatct agacccagct ttcttgtaca
     5221 aagtggttga tatccagcac agtggcggcc gctcgagtct agagggcccg cggttcgaat
     5281 acccatacga cgtcccagac tacgcttagt aatgattaat taaactagaa attctaccgg
     5341 gtaggggagg cgcttttccc aaggcagtct ggagtaaccc acccaagatc tggcctccgc
     5401 gccgggtttt ggcgcctccc gcgggcgccc ccctcctcac ggcgagccgc gtaagatctg
     5461 gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcctcacgg cgagcctagt
     5521 cgtgaggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
     5581 tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg
     5641 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
     5701 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa
     5761 gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt
     5821 gaattacttc cacctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg
     5881 ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg
     5941 cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg
     6001 ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt
     6061 tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg
     6121 gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc
     6181 tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg
     6241 tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg
     6301 caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat
     6361 ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct
     6421 ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc
     6481 tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg
     6541 cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga
     6601 tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc
     6661 agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgaagcggc cgccatgcta
     6721 gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacgga tccataactt
     6781 cgtataatgt atgctatacg aagttatgcc accatgtcta gactggacaa gagcaaagtc
     6841 ataaacggag ctctggaatt actcaatggt gtcggtatcg aaggcctgac gacaaggaaa
     6901 ctcgctcaaa agctgggagt tgagcagcct accctgtact ggcacgtgaa gaacaagcgg
     6961 gccctgctcg atgccctgcc aatcgagatg ctggacaggc atcataccca cttctgcccc
     7021 ctggaaggcg agtcatggca agactttctg cggaacaacg ccaagtcata ccgctgtgct
     7081 ctcctctcac atcgcgacgg ggctaaagtg catctcggca cccgcccaac agagaaacag
     7141 tacgaaaccc tggaaaatca gctcgcgttc ctgtgtcagc aaggcttctc cctggagaac
     7201 gcactgtacg ctctgtccgc cgtgggccac tttacactgg gctgcgtatt ggaggaacag
     7261 gagcatcaag tagcaaaaga ggaaagagag acacctacca ccgattctat gcccccactt
     7321 ctgagacaag caattgagct gttcgaccgg cagggagccg aacctgcctt ccttttcggc
     7381 ctggaactaa tcatatgtgg cctggagaaa cagctaaagt gcgaaagcgg cgggccgacc
     7441 gacgcccttg acgattttga cttagacatg ctcccagccg atgcccttga cgactttgac
     7501 cttgatatgc tgcctgctga cgctcttgac gattttgacc ttgacatgct ccccgggtaa
     7561 ctaagtaagg atccgcggcc gcactagagg aattccgccc ctctcccccc cccccctctc
     7621 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     7681 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     7741 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     7801 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     7861 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     7921 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     7981 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     8041 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
     8101 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     8161 ttttcctttg aaaaacacga tgataagctt accggtccac catggtgagc aagggcgagg
     8221 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     8281 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     8341 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     8401 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     8461 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     8521 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     8581 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     8641 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     8701 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     8761 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     8821 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     8881 ccgccgggat cactctcggc atggacgagc tgtacaagta agcatgcccg acggcgagga
     8941 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     9001 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     9061 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     9121 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     9181 cttctgatgt acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg
     9241 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     9301 tgaatgcaat tgttgttgtt tagtccctcc caattcgata tcaagcttat cgcgatagat
     9361 cctaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     9421 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     9481 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     9541 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     9601 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     9661 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     9721 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     9781 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     9841 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     9901 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctgagatc
     9961 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
    10021 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
    10081 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
    10141 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
    10201 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
    10261 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
    10321 agagtgagag gcccgggtta attaaggaaa gggctagatc attcttgaag acgaaagggc
    10381 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
    10441 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
    10501 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
    10561 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
    10621 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
    10681 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
    10741 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
    10801 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
    10861 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
    10921 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
    10981 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
    11041 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
    11101 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
    11161 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
    11221 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
    11281 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
    11341 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
    11401 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
    11461 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
    11521 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
    11581 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
    11641 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
    11701 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
    11761 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
    11821 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
    11881 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
    11941 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
    12001 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
    12061 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
    12121 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
    12181 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
    12241 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
    12301 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
    12361 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
    12421 tgcagcaagc tcatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
    12481 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
    12541 agctccccgt ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
    12601 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
    12661 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgacatgatt
    12721 acgaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
    12781 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc
    12841 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa
    12901 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
    12961 acaaacttag tagt
//
