pLV-mTurquoise2-Caspase-1-mNeonGreen

LOCUS        pLV-mTurquoise2-Caspase-1-mNeonGreen        9378 bp    DNA     circular SYN 14-APR-2025
DEFINITION  FRET analysis (pro caspase-1 cleavage) in living cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9378)
  AUTHORS   Magupalli VG, Negro R, Tian Y, Hauenstein AV, Di Caprio G, Skillern 
            W, Deng Q, Orning P, Alam HB, Maliga Z, Sharif H, Hu JJ, Evavold CL,
            Kagan JC, Schmidt FI, Fitzgerald KA, Kirchhausen T, Li Y, Wu H
  TITLE     HDAC6 mediates an aggresome-like mechanism for NLRP3 and pyrin 
            inflammasome activation.
  JOURNAL   Science. 2020 Sep 18;369(6510). pii: 369/6510/eaas8995. doi: 
            10.1126/science.aas8995.
  PUBMED    32943500
REFERENCE   2  (bases 1 to 9378)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science."; 
            date: "2020-09-18"; volume: "369(6510). pii: 369/6510/eaas8995. 
            doi"; pages: " 10.1126/science.aas8995"
COMMENT     Sequence Label: pLV-mTurquoise2-Caspase-1-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..9378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    846..1079
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /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    1575..1691
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        1714..2017
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2018..2221
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2172..2192
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2218..2242
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(2298..2300,2301..2303,2304..3014)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2298..2300,2301..2303,2304..3014)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2010)"
                     /label=mTurquoise
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2343..2364)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2604..2623)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2951..2972
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4278..4988
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      4533..4542
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    4995..5583
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5048..5068)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5466..5477)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5670..5903
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5975..6096
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6012..6031)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6066..6085
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6136..6271
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6291..6310)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6292..6310)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6320..6337)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6320..6336)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6329..6351)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6478..6933
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6565..6584)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6775..6796
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6959..7063
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7064..7132,7133..7924)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7282..7301)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8095..8683
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8584..8603
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8837..8854
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8971..8992
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9007..9012,9013..9030,9031..9037)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9045..9061
                     /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     9050..9072
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9069..9085
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9069..9085
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9104..9124
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9106..9124
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9152..9378
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     1501 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
     1561 cggtatcggt taacttttaa aagaaaaggg gggattgggg ggtatagtgc aggggaaaga
     1621 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     1681 ttcaaaattt tatcgataag cttgggagtt ccgcgttaca taacttacgg taaatggccc
     1741 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     1801 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     1861 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     1921 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     1981 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     2041 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     2101 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     2161 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     2221 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
     2281 aagacaccga ctctagaatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     2341 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     2401 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     2461 ccgtgccctg gcccaccctc gtgaccaccc tgtcctgggg cgtgcagtgc ttcgcccgct
     2521 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     2581 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     2641 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     2701 gcaacatcct ggggcacaag ctggagtaca actacttcag cgacaacgtc tatatcaccg
     2761 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg
     2821 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     2881 tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac cccaacgaga
     2941 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     3001 acgagctgta caagtccgga ctcagatctc gaggaatggc cgacaaggtc ctgaaggaga
     3061 agagaaagct gtttatccgt tccatgggtg aaggtacaat aaatggctta ctggatgaat
     3121 tattacagac aagggtgctg aacaaggaag agatggagaa agtaaaacgt gaaaatgcta
     3181 cagttatgga taagacccga gctttgattg actccgttat tccgaaaggg gcacaggcat
     3241 gccaaatttg catcacatac atttgtgaag aagacagtta cctggcaggg acgctgggac
     3301 tctcagcaga tcaaacatct ggaaattacc ttaatatgca agactctcaa ggagtacttt
     3361 cttcctttcc agctcctcag gcagtgcagg acaacccagc tatgcccaca tcctcaggct
     3421 cagaagggaa tgtcaagctt tgctccctag aagaagctca aaggatatgg aaacaaaagt
     3481 cggcagagat ttatccaata atggacaagt caagccgcac acgtcttgct ctcattatct
     3541 gcaatgaaga atttgacagt attcctagaa gaactggagc tgaggttgac atcacaggca
     3601 tgacaatgct gctacaaaat ctggggtaca gcgtagatgt gaaaaaaaat ctcactgctt
     3661 cggacatgac tacagagctg gaggcatttg cacaccgccc agagcacaag acctctgaca
     3721 gcacgttcct ggtgttcatg tctcatggta ttcgggaagg catttgtggg aagaaacact
     3781 ctgagcaagt cccagatata ctacaactca atgcaatctt taacatgttg aataccaaga
     3841 actgcccaag tttgaaggac aaaccgaagg tgatcatcat ccaggcctgc cgtggtgaca
     3901 gccctggtgt ggtgtggttt aaagattcag taggagtttc tggaaaccta tctttaccaa
     3961 ctacagaaga gtttgaggat gatgctatta agaaagccca catagagaag gattttatcg
     4021 ctttctgctc ttccacacca gataatgttt cttggagaca tcccacaatg ggctctgttt
     4081 ttattggaag actcattgaa catatgcaag aatatgcctg ttcctgtgat gtggaggaaa
     4141 ttttccgcaa ggttcgattt tcatttgagc agccagatgg tagagcgcag atgcccacca
     4201 ctgaaagagt gactttgaca agatgtttct acctcttccc aggacatgga agcggtggat
     4261 ccaccggtag cggctcaatg gtgagcaagg gcgaggagga taacatggcc tctctcccag
     4321 cgacacatga gttacacatc tttggctcca tcaacggtgt ggactttgac atggtgggtc
     4381 agggcaccgg caatccaaat gatggttatg aggagttaaa cctgaagtcc accaagggtg
     4441 acctccagtt ctccccctgg attctggtcc ctcatatcgg gtatggcttc catcagtacc
     4501 tgccctaccc tgacgggatg tcgcctttcc aggccgccat ggtagatggc tccggatacc
     4561 aagtccatcg cacaatgcag tttgaagatg gtgcctccct tactgttaac taccgctaca
     4621 cctacgaggg aagccacatc aaaggagagg cccaggtgaa ggggactggt ttccctgctg
     4681 acggtcctgt gatgaccaac tcgctgaccg ctgcggactg gtgcaggtcg aagaagactt
     4741 accccaacga caaaaccatc atcagtacct ttaagtggag ttacaccact ggaaatggca
     4801 agcgctaccg gagcactgcg cggaccacct acacctttgc caagccaatg gcggctaact
     4861 atctgaagaa ccagccgatg tacgtgttcc gtaagacgga gctcaagcac tccaagaccg
     4921 agctcaactt caaggagtgg caaaaggcct ttaccgatgt gatgggcatg gacgagctgt
     4981 acaagtaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     5041 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     5101 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     5161 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     5221 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     5281 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     5341 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     5401 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     5461 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     5521 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     5581 cgcctggaat tcgagctcgg tacctttaag accaatgact tacaaggcag ctgtagatct
     5641 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     5701 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     5761 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     5821 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     5881 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     5941 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
     6001 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6061 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
     6121 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     6181 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     6241 cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg
     6301 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     6361 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     6421 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
     6481 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     6541 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     6601 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     6661 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     6721 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     6781 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     6841 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     6901 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     6961 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     7021 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     7081 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     7141 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     7201 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     7261 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     7321 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     7381 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     7441 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     7501 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     7561 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     7621 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     7681 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     7741 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     7801 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     7861 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     7921 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     7981 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     8041 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     8101 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     8161 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     8221 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     8281 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     8341 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     8401 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     8461 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     8521 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     8581 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     8641 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     8701 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     8761 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     8821 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     8881 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     8941 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     9001 ccaggctttc cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     9061 atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta
     9121 aagggaacaa aagctggagc tgcaagctta atgtagtctt atgcaatact cttgtagtct
     9181 tgcaacatgg taacgatgag ttagcaacat gccttacaag gagagaaaaa gcaccgtgca
     9241 tgccgattgg tggaagtaag gtggtacgat cgtgccttat taggaaggca acagacgggt
     9301 ctgacatgga ttggacgaac cactgaattg ccgcattgca gagatattgt atttaagtgc
     9361 ctagctcgat acataaac
//