LOCUS       sequence-332719-       11729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet/Dox-inducible mammalian expression of CARD8 with a site III 
            mutation (F370G) with sgRNA binding site mutations.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11729)
  AUTHORS   Sharif H, Hollingsworth LR, Griswold AR, Hsiao JC, Wang Q, 
            Bachovchin DA, Wu H
  TITLE     Dipeptidyl peptidase 9 sets a threshold for CARD8 inflammasome 
            formation by sequestering its active C-terminal fragment.
  JOURNAL   Immunity. 2021 Jul 13;54(7):1392-1404.e10. doi: 
            10.1016/j.immuni.2021.04.024. Epub 2021 May 20.
  PUBMED    34019797
REFERENCE   2  (bases 1 to 11729)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.immuni.2021.04.024"; journalName: "Immunity"; date: 
            "2021-07-13- 13"; volume: "54"; issue: "7"; pages: "1392-1404.e10"
COMMENT     Sequence Label: pInducer20 CARD8-HA PAM F370G
FEATURES             Location/Qualifiers
     source          1..11729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    8..581
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(175..192)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     402..421
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             597..1391
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             597..1391
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(651..670)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1261..1280
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    1568..2156
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1621..1641)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2039..2050)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2231..2464
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(2583..2601)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2669..2773
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2774..2842,2843..3634)
                     /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(2992..3011)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3805..4393
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4294..4313
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4547..4564
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4804..4825
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4840..4845,4846..4863,4864..4870)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4878..4894
                     /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     4883..4905
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4902..4918
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4902..4918
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4968..4987
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             5179..5812
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5859..5984
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6481..6714
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6899..6943
                     /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    7236..7353
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    7408..7426
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7450..7468
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7492..7510
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7534..7552
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7576..7594
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7618..7636
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        7669..7707
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7704..7728
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    7837..7861
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(9479..9503)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             9556..9582
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        9668..10879
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     10716..10735
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(10938..11555,11556..11561,11562..11597,11598..11600,
                     11601..11636,11637..11639,11640..11675,11676..11684)
                     /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"
ORIGIN
        1 cccccccccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
       61 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
      121 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
      181 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
      241 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
      301 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
      361 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
      421 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
      481 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
      541 accacgggga cgtggttttc ctttgaaaaa cacgatgata agcttaccgg tccaccatga
      601 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
      661 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
      721 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
      781 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
      841 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
      901 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
      961 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     1021 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     1081 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     1141 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     1201 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     1261 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     1321 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     1381 agttcttctg atgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca
     1441 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     1501 aaaatgaatg caattgttgt tgtttagtcc ctcccaattc gatatcaagc ttatcgcgat
     1561 agatcctaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     1621 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     1681 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     1741 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     1801 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     1861 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     1921 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     1981 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     2041 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     2101 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctga
     2161 gatcctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     2221 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     2281 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     2341 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     2401 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     2461 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     2521 tcagagagtg agaggcccgg gttaattaag gaaagggcta gatcattctt gaagacgaaa
     2581 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     2641 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2701 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2761 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     2821 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     2881 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     2941 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3001 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     3061 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3121 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3181 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3241 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3301 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3361 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3421 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3481 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3541 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3601 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3661 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3721 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3781 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3841 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3901 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3961 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4021 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4081 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4141 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4201 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4261 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4321 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4381 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4441 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     4501 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     4561 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     4621 ttaatgcagc aagctcatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4681 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4741 aaaaagctcc ccgtggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     4801 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     4861 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacat
     4921 gattacgaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4981 actcatcaat gtatcttatc atgtctggat caactggata actcaagcta accaaaatca
     5041 tcccaaactt cccaccccat accctattac cactgccaat tacctgtggt ttcatttact
     5101 ctaaacctgt gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg
     5161 tactacaaac ttagtagttg gaagggctaa ttcactccca aagaagacaa gatatccttg
     5221 atctgtggat ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc
     5281 caggggtcag atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag
     5341 ataaggtaga agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc
     5401 atgggatgga tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat
     5461 ttcatcacgt ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt
     5521 gctacaaggg actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg
     5581 agtggcgagc cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc
     5641 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     5701 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     5761 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     5821 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     5881 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     5941 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     6001 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     6061 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     6121 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     6181 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     6241 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     6301 gtaagaccac cgcacagcaa gcggccggcc gctgatcttc agacctggag gaggagatat
     6361 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     6421 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     6481 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     6541 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     6601 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     6661 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     6721 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     6781 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     6841 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     6901 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     6961 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     7021 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     7081 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
     7141 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta
     7201 tcgccgaatt cacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     7261 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     7321 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     7381 ccagtttgga ctaggatcct ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
     7441 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca
     7501 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa
     7561 agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
     7621 ctatcagtga tagagaaaag tgaaagtcga gctcggtacc cgggtcgagg taggcgtgta
     7681 cggtgggagg cctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc
     7741 catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct ccgcggcccc
     7801 gaactagtcc agtgtggtgg aattctgcag atatcaacaa gtttgtacaa aaaagcaggc
     7861 ttcgccatgg aaaaaaagga gtgtccagaa aagagtagca gcagtgagga agagctgccg
     7921 agacgggaca gtggatccag taggaacata gatgcatcca aactcattag actacaagga
     7981 tcacggaaac tgttggttga caatagcata cgggaactgc aatacacaaa aactggaatt
     8041 ttttttcagg ctgaggcctg tgtgacaaat gatacggtat acagggagct accctgtgtt
     8101 tctgagaccc tttgtgacat ctcacatttt ttccaagaag atgatgagac agaggcagag
     8161 ccattattgt tccgtgctgt tcctgagtgt caactatctg ggggggacat tcccagtgta
     8221 tcagaagagc aggaatcttc agagggacaa gattcaggag acatttgctc agaagagaat
     8281 caaatagttt cctcttatgc ttctaaagtc tgttttgaga tcgaagaaga ttataaaaat
     8341 cgtcagtttc tggggcctga aggaaatgtg gatgttgagt tgattgataa gagcacaaac
     8401 agatacagcg tttggttccc cactgctggc tggtatctgt ggtcagccac aggcctcggc
     8461 ttcctggtaa gggatgaggt cacagttacc atcgccttcg gcagctggag tcagcacctg
     8521 gccctggacc tgcagcacca tgaacagtgg ctggtgggcg gccccttgtt tgatgtcact
     8581 gcagagccag aggaggctgt cgccgaaatc cacctccccc acttcatctc cctccaagca
     8641 ggtgaggtgg acgtctcctg gtttctcgtt gcccatttta agaatgaagg gatggtcctg
     8701 gagcatccag cccgggtgga gcctttctat gctgtcctgg aaagccccag cttctctctg
     8761 atgggcatcc tgctgcggat cgccagtggg actcgcctct ccatccccat cacttccaac
     8821 acattgatct attatcaccc ccaccccgaa gatattaagt tccacttgta ccttgtcccc
     8881 agcgacgcct tgctaacaaa ggcgatagat gatgaggaag atcgcttcca tggtgtgcgc
     8941 ctgcagactt cgcccccaat ggaacccctg aacggtggtt ccagttatat tgtgtctaat
     9001 tctgctaacc tgaaagtaat gcccaaggag ttgaaattgt cctacaggag ccctggagaa
     9061 attcagcact tctcaaaatt ctatgctggg cagatgaagg aacccattca acttgagatt
     9121 actgaaaaaa gacatgggac tttggtgtgg gatactgagg tgaagccagt ggatctccag
     9181 cttgtagctg catcagcccc tcctcctttc tcaggtgcag cctttgtgaa ggagaaccac
     9241 cggcaactcc aagccaggat gggggacctg aaaggggtgc tcgatgatct ccaagacaat
     9301 gaggttctta ctgagaatga gaaggagctg gtggagcagg aaaagacacg gcagagcaag
     9361 aatgaggcct tgctgagcat ggtggagaag aaaggggacc tggccctgga cgtgctcttc
     9421 agaagcatta gtgaaaggga cccttacctc gtgtcctatc ttagacagca gaatttggac
     9481 ccagctttct tgtacaaagt ggttgatatc cagcacagtg gcggccgctc gagtctagag
     9541 ggcccgcggt tcgaataccc atacgacgtc ccagactacg cttagtaatg attaattaaa
     9601 ctagaaattc taccgggtag gggaggcgct tttcccaagg cagtctggag taacccaccc
     9661 aagatctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cctcacggcg
     9721 agcgctgcca cgtcagacga agggcgcagc gagcgtcctg atccttccgc ccggacgctc
     9781 aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac
     9841 attttaggac gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca
     9901 ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa
     9961 cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga
    10021 tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg
    10081 ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag
    10141 agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact gggggttggg
    10201 gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg
    10261 gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc caaggtcttg
    10321 aggccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggg caccatctgg
    10381 ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg
    10441 cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg
    10501 tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc acctgccggt
    10561 aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct
    10621 cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct
    10681 ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg
    10741 ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt
    10801 tgggtcaata tgtaattttc agtgttagac tagtaaattg tccgctaaat tctggccgtt
    10861 tttggctttt ttgttagacg aagcttggta ccgagctcgg atctccaccc cgtaccggtc
    10921 ctgcagtcga attcaccatg tctagactgg acaagagcaa agtcataaac ggagctctgg
    10981 aattactcaa tggtgtcggt atcgaaggcc tgacgacaag gaaactcgct caaaagctgg
    11041 gagttgagca gcctaccctg tactggcacg tgaagaacaa gcgggccctg ctcgatgccc
    11101 tgccaatcga gatgctggac aggcatcata cccacttctg ccccctggaa ggcgagtcat
    11161 ggcaagactt tctgcggaac aacgccaagt cataccgctg tgctctcctc tcacatcgcg
    11221 acggggctaa agtgcatctc ggcacccgcc caacagagaa acagtacgaa accctggaaa
    11281 atcagctcgc gttcctgtgt cagcaaggct tctccctgga gaacgcactg tacgctctgt
    11341 ccgccgtggg ccactttaca ctgggctgcg tattggagga acaggagcat caagtagcaa
    11401 aagaggaaag agagacacct accaccgatt ctatgccccc acttctgaga caagcaattg
    11461 agctgttcga ccggcaggga gccgaacctg ccttcctttt cggcctggaa ctaatcatat
    11521 gtggcctgga gaaacagcta aagtgcgaaa gcggcgggcc gaccgacgcc cttgacgatt
    11581 ttgacttaga catgctccca gccgatgccc ttgacgactt tgaccttgat atgctgcctg
    11641 ctgacgctct tgacgatttt gaccttgaca tgctccccgg gtaactaagt aaggatccgc
    11701 ggccgcacta gaggaattcc gcccctctc
//
