LOCUS       sequence-332786-       11772 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet/Dox-inducible mammalian expression of CARD8 with sgRNA binding 
            site mutations.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11772)
  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 11772)
  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
FEATURES             Location/Qualifiers
     source          1..11772
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(157..174)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     384..403
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             579..1373
                     /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             579..1373
                     /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(633..652)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1243..1262
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    1550..2138
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1603..1623)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2021..2032)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2213..2446
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(2565..2583)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2651..2755
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2756..2824,2825..3616)
                     /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(2974..2993)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3787..4375
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4276..4295
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4529..4546
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4786..4807
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4822..4827,4828..4845,4846..4852)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4860..4876
                     /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     4865..4887
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4884..4900
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4884..4900
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4950..4969
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             5161..5794
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5841..5966
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6463..6696
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6881..6925
                     /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    7218..7335
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    join(7390..7408,7409..7431,7432..7450,7451..7473,7474..7492,
                     7493..7515,7516..7534,7535..7557,7558..7576,7577..7599,
                     7600..7618,7619..7641,7642..7660)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        7693..7731
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7728..7752
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    7861..7885
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(9503..9527)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             9580..9606
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        9692..10903
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     10740..10759
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(10962..11579,11580..11585,11586..11621,11622..11624,
                     11625..11660,11661..11663,11664..11699,11700..11708)
                     /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    join(11768..11772,1..563)
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
ORIGIN
        1 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
       61 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
      121 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
      181 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
      241 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
      301 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
      361 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
      421 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
      481 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
      541 tcctttgaaa aacacgatga taagcttacc ggtccaccat gattgaacaa gatggattgc
      601 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
      661 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
      721 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
      781 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
      841 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
      901 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
      961 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     1021 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     1081 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     1141 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     1201 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     1261 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     1321 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgatgtacaa
     1381 gtaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt
     1441 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     1501 gttgtttagt ccctcccaat tcgatatcaa gcttatcgcg atagatccta atcaacctct
     1561 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     1621 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     1681 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     1741 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     1801 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     1861 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     1921 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     1981 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     2041 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     2101 tcagacgagt cggatctccc tttgggccgc ctccccgcct gagatccttt aagaccaatg
     2161 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     2221 ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta
     2281 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     2341 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     2401 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt
     2461 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggccc
     2521 gggttaatta aggaaagggc tagatcattc ttgaagacga aagggcctcg tgatacgcct
     2581 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     2641 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2701 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2761 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2821 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2881 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2941 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     3001 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3061 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3121 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3181 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3241 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3301 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3361 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3421 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3481 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3541 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3601 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3661 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3721 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3781 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3841 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3901 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3961 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4021 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4081 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4141 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4201 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4261 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4321 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4381 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4441 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4501 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4561 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gcaagctcat
     4621 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4681 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct ccccgtggca
     4741 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     4801 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     4861 tgtgagcgga taacaatttc acacaggaaa cagctatgac atgattacga atttcacaaa
     4921 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4981 tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc
     5041 ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc
     5101 tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
     5161 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
     5221 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
     5281 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
     5341 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
     5401 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
     5461 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
     5521 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
     5581 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
     5641 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     5701 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     5761 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
     5821 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
     5881 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
     5941 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
     6001 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
     6061 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
     6121 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     6181 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     6241 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     6301 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     6361 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     6421 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     6481 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     6541 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     6601 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     6661 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     6721 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     6781 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     6841 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     6901 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     6961 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     7021 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     7081 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     7141 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     7201 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     7261 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     7321 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactaggatc
     7381 ctttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca
     7441 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa
     7501 agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
     7561 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
     7621 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagctcgg
     7681 tacccgggtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg
     7741 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
     7801 gaccgatcca gcctccgcgg ccccgaacta gtccagtgtg gtggaattct gcagatatca
     7861 acaagtttgt acaaaaaagc aggcttcgcc atggaaaaaa aggagtgtcc agaaaagagt
     7921 agcagcagtg aggaagagct gccgagacgg gacagtggat ccagtaggaa catagatgca
     7981 tccaaactca ttagactaca aggatcacgg aaactgttgg ttgacaatag catacgggaa
     8041 ctgcaataca caaaaactgg aatttttttt caggctgagg cctgtgtgac aaatgatacg
     8101 gtatacaggg agctaccctg tgtttctgag accctttgtg acatctcaca ttttttccaa
     8161 gaagatgatg agacagaggc agagccatta ttgttccgtg ctgttcctga gtgtcaacta
     8221 tctggggggg acattcccag tgtatcagaa gagcaggaat cttcagaggg acaagattca
     8281 ggagacattt gctcagaaga gaatcaaata gtttcctctt atgcttctaa agtctgtttt
     8341 gagatcgaag aagattataa aaatcgtcag tttctggggc ctgaaggaaa tgtggatgtt
     8401 gagttgattg ataagagcac aaacagatac agcgtttggt tccccactgc tggctggtat
     8461 ctgtggtcag ccacaggcct cggcttcctg gtaagggatg aggtcacagt taccatcgcc
     8521 ttcggcagct ggagtcagca cctggccctg gacctgcagc accatgaaca gtggctggtg
     8581 ggcggcccct tgtttgatgt cactgcagag ccagaggagg ctgtcgccga aatccacctc
     8641 ccccacttca tctccctcca agcaggtgag gtggacgtct cctggtttct cgttgcccat
     8701 tttaagaatg aagggatggt cctggagcat ccagcccggg tggagccttt ctatgctgtc
     8761 ctggaaagcc ccagcttctc tctgatgggc atcctgctgc ggatcgccag tgggactcgc
     8821 ctctccatcc ccatcacttc caacacattg atctattatc acccccaccc cgaagatatt
     8881 aagttccact tgtaccttgt ccccagcgac gccttgctaa caaaggcgat agatgatgag
     8941 gaagatcgct tccatggtgt gcgcctgcag acttcgcccc caatggaacc cctgaacttt
     9001 ggttccagtt atattgtgtc taattctgct aacctgaaag taatgcccaa ggagttgaaa
     9061 ttgtcctaca ggagccctgg agaaattcag cacttctcaa aattctatgc tgggcagatg
     9121 aaggaaccca ttcaacttga gattactgaa aaaagacatg ggactttggt gtgggatact
     9181 gaggtgaagc cagtggatct ccagcttgta gctgcatcag cccctcctcc tttctcaggt
     9241 gcagcctttg tgaaggagaa ccaccggcaa ctccaagcca ggatggggga cctgaaaggg
     9301 gtgctcgatg atctccaaga caatgaggtt cttactgaga atgagaagga gctggtggag
     9361 caggaaaaga cacggcagag caagaatgag gccttgctga gcatggtgga gaagaaaggg
     9421 gacctggccc tggacgtgct cttcagaagc attagtgaaa gggaccctta cctcgtgtcc
     9481 tatcttagac agcagaattt ggacccagct ttcttgtaca aagtggttga tatccagcac
     9541 agtggcggcc gctcgagtct agagggcccg cggttcgaat acccatacga cgtcccagac
     9601 tacgcttagt aatgattaat taaactagaa attctaccgg gtaggggagg cgcttttccc
     9661 aaggcagtct ggagtaaccc acccaagatc tggcctccgc gccgggtttt ggcgcctccc
     9721 gcgggcgccc ccctcctcac ggcgagcgct gccacgtcag acgaagggcg cagcgagcgt
     9781 cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta
     9841 gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg
     9901 gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg
     9961 gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca
    10021 cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat
    10081 cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc
    10141 tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa
    10201 ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt
    10261 gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt
    10321 gggcggcaag aacccaaggt cttgaggcct tcgctaatgc gggaaagctc ttattcgggt
    10381 gagatgggct ggggcaccat ctggggaccc tgacgtgaag tttgtcactg actggagaac
    10441 tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt gccgttgggc agtgcacccg
    10501 tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa
    10561 tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct tttctccgtc gcaggacgca
    10621 gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg ccggacctct ggtgagggga
    10681 gggataagtg aggcgtcagt ttctttggtc ggttttatgt acctatcttc ttaagtagct
    10741 gaagctccgg ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg aagtttttta
    10801 ggcacctttt gaaatgtaat catttgggtc aatatgtaat tttcagtgtt agactagtaa
    10861 attgtccgct aaattctggc cgtttttggc ttttttgtta gacgaagctt ggtaccgagc
    10921 tcggatctcc accccgtacc ggtcctgcag tcgaattcac catgtctaga ctggacaaga
    10981 gcaaagtcat aaacggagct ctggaattac tcaatggtgt cggtatcgaa ggcctgacga
    11041 caaggaaact cgctcaaaag ctgggagttg agcagcctac cctgtactgg cacgtgaaga
    11101 acaagcgggc cctgctcgat gccctgccaa tcgagatgct ggacaggcat catacccact
    11161 tctgccccct ggaaggcgag tcatggcaag actttctgcg gaacaacgcc aagtcatacc
    11221 gctgtgctct cctctcacat cgcgacgggg ctaaagtgca tctcggcacc cgcccaacag
    11281 agaaacagta cgaaaccctg gaaaatcagc tcgcgttcct gtgtcagcaa ggcttctccc
    11341 tggagaacgc actgtacgct ctgtccgccg tgggccactt tacactgggc tgcgtattgg
    11401 aggaacagga gcatcaagta gcaaaagagg aaagagagac acctaccacc gattctatgc
    11461 ccccacttct gagacaagca attgagctgt tcgaccggca gggagccgaa cctgccttcc
    11521 ttttcggcct ggaactaatc atatgtggcc tggagaaaca gctaaagtgc gaaagcggcg
    11581 ggccgaccga cgcccttgac gattttgact tagacatgct cccagccgat gcccttgacg
    11641 actttgacct tgatatgctg cctgctgacg ctcttgacga ttttgacctt gacatgctcc
    11701 ccgggtaact aagtaaggat ccgcggccgc actagaggaa ttccgcccct ctcccccccc
    11761 ccccntctcc ct
//
