LOCUS       sequence-332777-       11729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet/Dox-inducible mammalian expression of CARD8 with a site III 
            mutation (F405G) 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 F405G
FEATURES             Location/Qualifiers
     source          1..11729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(159..176)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     386..405
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             581..1375
                     /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             581..1375
                     /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(635..654)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1245..1264
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    1552..2140
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1605..1625)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2023..2034)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2215..2448
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(2567..2585)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2653..2757
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2758..2826,2827..3618)
                     /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(2976..2995)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3789..4377
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4278..4297
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4531..4548
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4788..4809
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4824..4829,4830..4847,4848..4854)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4862..4878
                     /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     4867..4889
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4886..4902
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4886..4902
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4952..4971
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             5163..5796
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5843..5968
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6465..6698
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6883..6927
                     /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    7220..7337
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    7392..7410
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7434..7452
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7476..7494
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7518..7536
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7560..7578
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7602..7620
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        7653..7691
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7688..7712
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    7821..7845
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(9463..9487)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             9540..9566
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        9652..10863
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     10700..10719
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(10922..11539,11540..11545,11546..11581,11582..11584,
                     11585..11620,11621..11623,11624..11659,11660..11668)
                     /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(11721..11729,1..565)
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
ORIGIN
        1 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
       61 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
      121 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
      181 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
      241 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
      301 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
      361 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
      421 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
      481 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
      541 tttcctttga aaaacacgat gataagctta ccggtccacc atgattgaac aagatggatt
      601 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
      661 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
      721 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
      781 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
      841 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
      901 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
      961 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     1021 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     1081 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     1141 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     1201 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     1261 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     1321 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgatgtac
     1381 aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
     1441 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     1501 ttgttgttta gtccctccca attcgatatc aagcttatcg cgatagatcc taatcaacct
     1561 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     1621 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     1681 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     1741 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     1801 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     1861 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     1921 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     1981 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     2041 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     2101 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctgagatcct ttaagaccaa
     2161 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     2221 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     2281 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     2341 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     2401 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     2461 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagaggc
     2521 ccgggttaat taaggaaagg gctagatcat tcttgaagac gaaagggcct cgtgatacgc
     2581 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     2641 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2701 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     2761 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     2821 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     2881 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     2941 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3001 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3061 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3121 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3181 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3241 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3301 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3361 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3421 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3481 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3541 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3601 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     3661 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3721 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3781 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3841 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3901 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3961 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4021 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4081 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4141 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4201 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4261 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4321 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4381 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4441 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     4501 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     4561 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagcaagctc
     4621 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4681 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctccccgtgg
     4741 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     4801 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     4861 attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac gaatttcaca
     4921 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4981 tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc
     5041 ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac ctgtgattcc
     5101 tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta
     5161 gttggaaggg ctaattcact cccaaagaag acaagatatc cttgatctgt ggatctacca
     5221 cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc
     5281 actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc
     5341 caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc
     5401 ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg
     5461 agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc
     5521 cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag
     5581 atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct
     5641 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5701 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5761 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     5821 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     5881 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     5941 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     6001 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     6061 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     6121 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     6181 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     6241 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     6301 gcaagcggcc ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga
     6361 agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag
     6421 gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt
     6481 gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag
     6541 gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag
     6601 gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc
     6661 ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga
     6721 aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa
     6781 cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc
     6841 ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta
     6901 ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg
     6961 tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct
     7021 gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac
     7081 ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga
     7141 gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgccg aattcacaaa
     7201 tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg
     7261 aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta
     7321 caaaaattca aaattttcgg gtttattaca gggacagcag agatccagtt tggactagga
     7381 tcctttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat
     7441 cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg
     7501 aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact
     7561 ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga
     7621 aaagtgaaag tcgagctcgg tacccgggtc gaggtaggcg tgtacggtgg gaggcctata
     7681 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
     7741 acctccatag aagacaccgg gaccgatcca gcctccgcgg ccccgaacta gtccagtgtg
     7801 gtggaattct gcagatatca acaagtttgt acaaaaaagc aggcttcgcc atggaaaaaa
     7861 aggagtgtcc agaaaagagt agcagcagtg aggaagagct gccgagacgg gacagtggat
     7921 ccagtaggaa catagatgca tccaaactca ttagactaca aggatcacgg aaactgttgg
     7981 ttgacaatag catacgggaa ctgcaataca caaaaactgg aatttttttt caggctgagg
     8041 cctgtgtgac aaatgatacg gtatacaggg agctaccctg tgtttctgag accctttgtg
     8101 acatctcaca ttttttccaa gaagatgatg agacagaggc agagccatta ttgttccgtg
     8161 ctgttcctga gtgtcaacta tctggggggg acattcccag tgtatcagaa gagcaggaat
     8221 cttcagaggg acaagattca ggagacattt gctcagaaga gaatcaaata gtttcctctt
     8281 atgcttctaa agtctgtttt gagatcgaag aagattataa aaatcgtcag tttctggggc
     8341 ctgaaggaaa tgtggatgtt gagttgattg ataagagcac aaacagatac agcgtttggt
     8401 tccccactgc tggctggtat ctgtggtcag ccacaggcct cggcttcctg gtaagggatg
     8461 aggtcacagt taccatcgcc ttcggcagct ggagtcagca cctggccctg gacctgcagc
     8521 accatgaaca gtggctggtg ggcggcccct tgtttgatgt cactgcagag ccagaggagg
     8581 ctgtcgccga aatccacctc ccccacttca tctccctcca agcaggtgag gtggacgtct
     8641 cctggtttct cgttgcccat tttaagaatg aagggatggt cctggagcat ccagcccggg
     8701 tggagccttt ctatgctgtc ctggaaagcc ccagcttctc tctgatgggc atcctgctgc
     8761 ggatcgccag tgggactcgc ctctccatcc ccatcacttc caacacattg atctattatc
     8821 acccccaccc cgaagatatt aagttccact tgtaccttgt ccccagcgac gccttgctaa
     8881 caaaggcgat agatgatgag gaagatcgct tccatggtgt gcgcctgcag acttcgcccc
     8941 caatggaacc cctgaacttt ggttccagtt atattgtgtc taattctgct aacctgaaag
     9001 taatgcccaa ggagttgaaa ttgtcctaca ggagccctgg agaaattcag cacttctcaa
     9061 aaggctatgc tgggcagatg aaggaaccca ttcaacttga gattactgaa aaaagacatg
     9121 ggactttggt gtgggatact gaggtgaagc cagtggatct ccagcttgta gctgcatcag
     9181 cccctcctcc tttctcaggt gcagcctttg tgaaggagaa ccaccggcaa ctccaagcca
     9241 ggatggggga cctgaaaggg gtgctcgatg atctccaaga caatgaggtt cttactgaga
     9301 atgagaagga gctggtggag caggaaaaga cacggcagag caagaatgag gccttgctga
     9361 gcatggtgga gaagaaaggg gacctggccc tggacgtgct cttcagaagc attagtgaaa
     9421 gggaccctta cctcgtgtcc tatcttagac agcagaattt ggacccagct ttcttgtaca
     9481 aagtggttga tatccagcac agtggcggcc gctcgagtct agagggcccg cggttcgaat
     9541 acccatacga cgtcccagac tacgcttagt aatgattaat taaactagaa attctaccgg
     9601 gtaggggagg cgcttttccc aaggcagtct ggagtaaccc acccaagatc tggcctccgc
     9661 gccgggtttt ggcgcctccc gcgggcgccc ccctcctcac ggcgagcgct gccacgtcag
     9721 acgaagggcg cagcgagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct
     9781 gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact
     9841 tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt
     9901 cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata
     9961 aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct
    10021 tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc
    10081 tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg
    10141 tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat
    10201 ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga
    10261 aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgaggcct tcgctaatgc
    10321 gggaaagctc ttattcgggt gagatgggct ggggcaccat ctggggaccc tgacgtgaag
    10381 tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt
    10441 gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca
    10501 cccgttctgt tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct
    10561 tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg
    10621 ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt
    10681 acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt
    10741 gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat
    10801 tttcagtgtt agactagtaa attgtccgct aaattctggc cgtttttggc ttttttgtta
    10861 gacgaagctt ggtaccgagc tcggatctcc accccgtacc ggtcctgcag tcgaattcac
    10921 catgtctaga ctggacaaga gcaaagtcat aaacggagct ctggaattac tcaatggtgt
    10981 cggtatcgaa ggcctgacga caaggaaact cgctcaaaag ctgggagttg agcagcctac
    11041 cctgtactgg cacgtgaaga acaagcgggc cctgctcgat gccctgccaa tcgagatgct
    11101 ggacaggcat catacccact tctgccccct ggaaggcgag tcatggcaag actttctgcg
    11161 gaacaacgcc aagtcatacc gctgtgctct cctctcacat cgcgacgggg ctaaagtgca
    11221 tctcggcacc cgcccaacag agaaacagta cgaaaccctg gaaaatcagc tcgcgttcct
    11281 gtgtcagcaa ggcttctccc tggagaacgc actgtacgct ctgtccgccg tgggccactt
    11341 tacactgggc tgcgtattgg aggaacagga gcatcaagta gcaaaagagg aaagagagac
    11401 acctaccacc gattctatgc ccccacttct gagacaagca attgagctgt tcgaccggca
    11461 gggagccgaa cctgccttcc ttttcggcct ggaactaatc atatgtggcc tggagaaaca
    11521 gctaaagtgc gaaagcggcg ggccgaccga cgcccttgac gattttgact tagacatgct
    11581 cccagccgat gcccttgacg actttgacct tgatatgctg cctgctgacg ctcttgacga
    11641 ttttgacctt gacatgctcc ccgggtaact aagtaaggat ccgcggccgc actagaggaa
    11701 ttccgcccct ctcccccccc cccctctcc
//
