LOCUS       sequence-332788-       11771 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet/Dox-inducible mammalian expression of CARD8 with a site III 
            mutation (L368G) with sgRNA binding site mutations.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11771)
  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 11771)
  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 L368G
FEATURES             Location/Qualifiers
     source          1..11771
                     /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"
     protein_bind    complement(4343..4367)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4420..4446
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        4532..5743
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5580..5599
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(5802..6419,6420..6425,6426..6461,6462..6464,6465..6500,
                     6501..6503,6504..6539,6540..6548)
                     /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    6601..7174
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6768..6785)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6995..7014
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7190..7984
                     /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             7190..7984
                     /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(7244..7263)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7854..7873
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    8161..8749
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8214..8234)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8632..8643)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8824..9057
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9176..9194)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9262..9366
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9367..9435,9436..10227)
                     /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(9585..9604)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10398..10986
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10887..10906
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11140..11157
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11397..11418
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11433..11438,11439..11456,11457..11463)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11471..11487
                     /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     11476..11498
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11495..11511
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11495..11511
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11561..11580
                     /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 aggcttcgcc atggaaaaaa aggagtgtcc agaaaagagt
     2761 agcagcagtg aggaagagct gccgagacgg gacagtggat ccagtaggaa catagatgca
     2821 tccaaactca ttagactaca aggatcacgg aaactgttgg ttgacaatag catacgggaa
     2881 ctgcaataca caaaaactgg aatttttttt caggctgagg cctgtgtgac aaatgatacg
     2941 gtatacaggg agctaccctg tgtttctgag accctttgtg acatctcaca ttttttccaa
     3001 gaagatgatg agacagaggc agagccatta ttgttccgtg ctgttcctga gtgtcaacta
     3061 tctggggggg acattcccag tgtatcagaa gagcaggaat cttcagaggg acaagattca
     3121 ggagacattt gctcagaaga gaatcaaata gtttcctctt atgcttctaa agtctgtttt
     3181 gagatcgaag aagattataa aaatcgtcag tttctggggc ctgaaggaaa tgtggatgtt
     3241 gagttgattg ataagagcac aaacagatac agcgtttggt tccccactgc tggctggtat
     3301 ctgtggtcag ccacaggcct cggcttcctg gtaagggatg aggtcacagt taccatcgcc
     3361 ttcggcagct ggagtcagca cctggccctg gacctgcagc accatgaaca gtggctggtg
     3421 ggcggcccct tgtttgatgt cactgcagag ccagaggagg ctgtcgccga aatccacctc
     3481 ccccacttca tctccctcca agcaggtgag gtggacgtct cctggtttct cgttgcccat
     3541 tttaagaatg aagggatggt cctggagcat ccagcccggg tggagccttt ctatgctgtc
     3601 ctggaaagcc ccagcttctc tctgatgggc atcctgctgc ggatcgccag tgggactcgc
     3661 ctctccatcc ccatcacttc caacacattg atctattatc acccccaccc cgaagatatt
     3721 aagttccact tgtaccttgt ccccagcgac gccttgctaa caaaggcgat agatgatgag
     3781 gaagatcgct tccatggtgt gcgcctgcag acttcgcccc caatggaacc cgggaacttt
     3841 ggttccagtt atattgtgtc taattctgct aacctgaaag taatgcccaa ggagttgaaa
     3901 ttgtcctaca ggagccctgg agaaattcag cacttctcaa aattctatgc tgggcagatg
     3961 aaggaaccca ttcaacttga gattactgaa aaaagacatg ggactttggt gtgggatact
     4021 gaggtgaagc cagtggatct ccagcttgta gctgcatcag cccctcctcc tttctcaggt
     4081 gcagcctttg tgaaggagaa ccaccggcaa ctccaagcca ggatggggga cctgaaaggg
     4141 gtgctcgatg atctccaaga caatgaggtt cttactgaga atgagaagga gctggtggag
     4201 caggaaaaga cacggcagag caagaatgag gccttgctga gcatggtgga gaagaaaggg
     4261 gacctggccc tggacgtgct cttcagaagc attagtgaaa gggaccctta cctcgtgtcc
     4321 tatcttagac agcagaattt ggacccagct ttcttgtaca aagtggttga tatccagcac
     4381 agtggcggcc gctcgagtct agagggcccg cggttcgaat acccatacga cgtcccagac
     4441 tacgcttagt aatgattaat taaactagaa attctaccgg gtaggggagg cgcttttccc
     4501 aaggcagtct ggagtaaccc acccaagatc tggcctccgc gccgggtttt ggcgcctccc
     4561 gcgggcgccc ccctcctcac ggcgagcgct gccacgtcag acgaagggcg cagcgagcgt
     4621 cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta
     4681 gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg
     4741 gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg
     4801 gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca
     4861 cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat
     4921 cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc
     4981 tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa
     5041 ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt
     5101 gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt
     5161 gggcggcaag aacccaaggt cttgaggcct tcgctaatgc gggaaagctc ttattcgggt
     5221 gagatgggct ggggcaccat ctggggaccc tgacgtgaag tttgtcactg actggagaac
     5281 tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt gccgttgggc agtgcacccg
     5341 tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa
     5401 tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct tttctccgtc gcaggacgca
     5461 gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg ccggacctct ggtgagggga
     5521 gggataagtg aggcgtcagt ttctttggtc ggttttatgt acctatcttc ttaagtagct
     5581 gaagctccgg ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg aagtttttta
     5641 ggcacctttt gaaatgtaat catttgggtc aatatgtaat tttcagtgtt agactagtaa
     5701 attgtccgct aaattctggc cgtttttggc ttttttgtta gacgaagctt ggtaccgagc
     5761 tcggatctcc accccgtacc ggtcctgcag tcgaattcac catgtctaga ctggacaaga
     5821 gcaaagtcat aaacggagct ctggaattac tcaatggtgt cggtatcgaa ggcctgacga
     5881 caaggaaact cgctcaaaag ctgggagttg agcagcctac cctgtactgg cacgtgaaga
     5941 acaagcgggc cctgctcgat gccctgccaa tcgagatgct ggacaggcat catacccact
     6001 tctgccccct ggaaggcgag tcatggcaag actttctgcg gaacaacgcc aagtcatacc
     6061 gctgtgctct cctctcacat cgcgacgggg ctaaagtgca tctcggcacc cgcccaacag
     6121 agaaacagta cgaaaccctg gaaaatcagc tcgcgttcct gtgtcagcaa ggcttctccc
     6181 tggagaacgc actgtacgct ctgtccgccg tgggccactt tacactgggc tgcgtattgg
     6241 aggaacagga gcatcaagta gcaaaagagg aaagagagac acctaccacc gattctatgc
     6301 ccccacttct gagacaagca attgagctgt tcgaccggca gggagccgaa cctgccttcc
     6361 ttttcggcct ggaactaatc atatgtggcc tggagaaaca gctaaagtgc gaaagcggcg
     6421 ggccgaccga cgcccttgac gattttgact tagacatgct cccagccgat gcccttgacg
     6481 actttgacct tgatatgctg cctgctgacg ctcttgacga ttttgacctt gacatgctcc
     6541 ccgggtaact aagtaaggat ccgcggccgc actagaggaa ttccgcccct ctcccccccc
     6601 cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
     6661 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
     6721 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
     6781 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
     6841 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
     6901 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
     6961 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
     7021 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
     7081 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
     7141 ggacgtggtt ttcctttgaa aaacacgatg ataagcttac cggtccacca tgattgaaca
     7201 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     7261 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     7321 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     7381 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     7441 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     7501 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     7561 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     7621 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     7681 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     7741 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     7801 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     7861 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     7921 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     7981 ctgatgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     8041 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     8101 atgcaattgt tgttgtttag tccctcccaa ttcgatatca agcttatcgc gatagatcct
     8161 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     8221 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     8281 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     8341 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     8401 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     8461 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     8521 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     8581 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     8641 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     8701 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tgagatcctt
     8761 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     8821 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
     8881 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     8941 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     9001 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     9061 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     9121 gtgagaggcc cgggttaatt aaggaaaggg ctagatcatt cttgaagacg aaagggcctc
     9181 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     9241 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     9301 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     9361 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     9421 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     9481 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     9541 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     9601 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     9661 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     9721 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     9781 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     9841 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     9901 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     9961 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
    10021 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
    10081 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
    10141 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
    10201 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
    10261 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
    10321 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
    10381 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
    10441 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
    10501 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
    10561 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
    10621 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
    10681 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
    10741 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
    10801 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
    10861 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
    10921 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
    10981 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
    11041 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
    11101 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
    11161 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
    11221 agcaagctca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
    11281 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
    11341 tccccgtggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
    11401 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
    11461 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg
    11521 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
    11581 aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa
    11641 cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc
    11701 tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca
    11761 aacttagtag t
//
