LOCUS       sequence-431940-       11471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Proximity biotinylation of msNLRP3(Delta-PYD, I125M-END).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11471)
  AUTHORS   Hollingsworth LR, Veeraraghavan P, Paulo JA, Harper JW
  TITLE     Spatiotemporal proteomic profiling of cellular responses to NLRP3 
            agonists
  JOURNAL   bioRxiv 2024.04.19.590338
REFERENCE   2  (bases 1 to 11471)
  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"; journalName: "bioRxiv 
            2024.04.19.590338"
COMMENT     Sequence Label: pLV UBC msNLRP3(Delta-PYD,
            I125M-END)-APEX2-3xFLAG-...
FEATURES             Location/Qualifiers
     source          1..11471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    155..271
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        274..672
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     regulatory      718..727
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3493..3502
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3499..4248
                     /codon_start=1
                     /product="soybean ascorbate peroxidase, engineered to serve
                     as a monomeric tag for electron microscopy and live-cell 
                     proteomics (Lam et al., 2015)"
                     /label=APEX2
                     /note="more active than the original APEX due to the A134P 
                     mutation"
                     /translation="MGKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAG
                     TFDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVA
                     VEVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGH
                     TIGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVD
                     KYAADEDAFFADYAEAHQKLSELGFADA"
     CDS             4267..4332
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    4369..4921
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4513..4530)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4740..4759
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4933..4935,4936..4938,4939..5652)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4933..4935,4936..4938,4939..5649)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5239..5258)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5586..5607
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5668..6256
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5721..5741)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6139..6150)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6343..6576
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6648..6769
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6685..6704)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6739..6758
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6809..6944
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6964..6983)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6965..6983)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6993..7010)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6993..7009)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7002..7024)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7151..7606
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7238..7257)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7448..7469
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7632..7736
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7737..7805,7806..8597)
                     /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(7955..7974)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8768..9356
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9257..9276
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9510..9527
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9644..9665
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9680..9685,9686..9703,9704..9710)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9718..9734
                     /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     9723..9745
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9742..9758
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9742..9758
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9777..9797
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9779..9797
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9825..10051
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             10052..10232
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    10279..10404
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10897..11130
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11315..11359
                     /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"
ORIGIN
        1 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
       61 ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc
      121 gattagtgaa cggatctcga cggtatcggt taacttttaa aagaaaaggg gggattgggg
      181 ggtatagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
      241 tacaaaaaca aattacaaaa ttcaaaattt tatggcctcc gcgccgggtt ttggcgcctc
      301 ccgcgggcgc ccccctcctc acggcgagcg ctgccacgtc agacgaaggg cgcagcgagc
      361 gtcctgatcc ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct
      421 tagaacccca gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac
      481 tggttttctt tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg
      541 cggagggatc tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg
      601 cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg
      661 atcgtcactt ggggagacgc catccacgct gttttgacct ccatagaaga caccgacgcc
      721 accatggatt actgtaagat gtacagacga catgtgagaa gcaggttcta ctctatcaag
      781 gacaggaacg cgcgtctagg tgagagtgtg gacctcaaca gtcgctacac gcagctccaa
      841 ctggtcaagg agcatccaag caagcaggag cgggagcatg aactcctgac catcggccgg
      901 actaaaatgc gggacagccc catgagttcc cttaagctgg agctgctgtt tgagcccgag
      961 gacgggcact cggagcctgt gcacacagtg gtgttccagg gagcagcagg catcgggaaa
     1021 accatcctag ccaggaagat tatgttggac tgggcactgg gaaagctctt caaagacaaa
     1081 tttgactatt tgttctttat ccactgccga gaggtgagcc tcaggacgcc aaggagtcta
     1141 gcagacctga ttgtcagctg ctggcctgac ccaaacccac cagtgtgcaa gatcctgcgc
     1201 aagccttcca ggatcctctt cctcatggat ggctttgatg agctacaagg ggcctttgac
     1261 gagcacattg gggaggtctg cacagactgg caaaaggctg tgcggggaga cattctgcta
     1321 agcagcctca tccgaaagaa actgctgccc aaggcctctc tgctcataac gacgaggccg
     1381 gtagccttgg agaaactgca gcatctcctg gaccaccccc gccatgtgga gatcctaggt
     1441 ttctctgagg ccaaaaggaa ggagtatttc tttaagtatt tctccaacga gctgcaggcc
     1501 cgggaggcct tcaggctgat ccaagagaat gaggtcctct ttaccatgtg cttcatcccc
     1561 ctggtctgct ggattgtgtg cacggggcta aagcaacaga tggagaccgg gaagagcctg
     1621 gcccagacct ccaagaccac tacggccgtc tacgtcttct tcctttccag cctgctgcaa
     1681 tcccgggggg gcattgagga gcatctcttc tctgactacc tacaggggct ctgttcactg
     1741 gctgcggatg gaatttggaa ccagaaaatc ctatttgagg agtgtgatct gcggaagcac
     1801 ggcctgcaga agactgacgt ctccgctttc ctgaggatga acgtgttcca gaaggaagtg
     1861 gactgcgaga gattctacag cttcagccac atgactttcc aggagttctt cgctgctatg
     1921 tactatttgc tggaagagga ggcagagggg gagaccgtga ggaaaggacc aggaggttgt
     1981 tcagatcttc tgaaccgaga cgtgaaggtc ctactagaaa attacggcaa gtttgaaaaa
     2041 ggctatctga tttttgttgt ccgattcctc tttggccttg taaaccagga gagaacctct
     2101 tatttggaga agaaactaag ttgcaagatc tctcagcaag tcagactgga actactgaag
     2161 tggattgaag tgaaagccaa ggccaagaag ctgcagtggc agcccagcca actggaactg
     2221 ttctactgcc tgtacgagat gcaggaggaa gactttgtgc agagtgccat ggaccacttt
     2281 cccaaaattg agatcaacct ctctaccaga atggaccacg tggtttcctc cttttgtatt
     2341 aagaactgtc atagggtcaa aacgctttcc ctgggttttt ttcacaactc gcccaaggag
     2401 gaagaagaag agaggagagg aggtcgaccc ttggaccagg ttcagtgtgt tttcccagac
     2461 actcatgttg cctgttcttc cagactggtg aactgctgcc tcacttctag cttctgccgt
     2521 ggtctcttct caagtctaag caccaaccgg agcctcactg aactggacct cagtgacaat
     2581 actctgggag acccgggcat gagggtgctg tgtgaggcac tccagcaccc aggctgtaac
     2641 attcagagac tgtggttggg gcgctgcgga ctgtcccatc aatgctgctt cgacatctcc
     2701 tctgtcctga gcagcagcca gaagctggtg gagctggacc tcagtgacaa tgccctgggg
     2761 gactttggaa tcagattgct gtgtgtggga ctgaagcacc tgctctgcaa cctccagaaa
     2821 ctgtggttgg tgagctgctg tctcacatcc gcgtgttgtc aggatctcgc attggttctg
     2881 agctccaacc attctctgac cagactgtac attggagaaa atgccttggg agactcagga
     2941 gtccaagttt tgtgtgaaaa gatgaaggac ccacagtgta acttgcagaa gctggggttg
     3001 gtgaattccg gccttacttc aatctgttgt tcagctctga cctctgtgct caaaaccaac
     3061 cagaacttca cacacctcta tctacgaagc aatgcccttg gagacacagg actcaggctc
     3121 ctctgtgagg ggcttctgca cccggactgt aaactacaga tgctggaatt agacaactgc
     3181 agcctcacct cacacagctg ctggaatctc tccacaattc tgacccacaa ccacagcctt
     3241 cggaagctga acctgggcaa caatgatctt ggcgatctgt gcgtggtgac cctctgtgag
     3301 gtgctgaaac agcagggctg cctcctgcag agcctacagt tgggtgaaat gtacttaaat
     3361 cgtgaaacaa aacgtgcctt agaagcgctc caggaagaaa agcctgagct gactatagtc
     3421 ttcgagattt cctggggcgg atccggggga agcggcggat ccatcgaaga cggaagagcg
     3481 gatccaccgg tcgccaccat gggaaagtct tacccaactg tgagtgctga ttaccaggac
     3541 gccgttgaga aggcgaagaa gaagctcaga ggcttcatcg ctgagaagag atgcgctcct
     3601 ctaatgctcc gtttggcatt ccactctgct ggaacctttg acaagggcac gaagaccggt
     3661 ggacccttcg gaaccatcaa gcaccctgcc gaactggctc acagcgctaa caacggtctt
     3721 gacatcgctg ttaggctttt ggagccactc aaggcggagt tccctatttt gagctacgcc
     3781 gatttctacc agttggctgg cgttgttgcc gttgaggtca cgggtggacc taaggttcca
     3841 ttccaccctg gaagagagga caagcctgag ccaccaccag agggtcgctt gcccgatccc
     3901 actaagggtt ctgaccattt gagagatgtg tttggcaaag ctatggggct tactgaccaa
     3961 gatatcgttg ctctatctgg gggtcacact attggagctg cacacaagga gcgttctgga
     4021 tttgagggtc cctggacctc taatcctctt attttcgaca actcatactt cacggagttg
     4081 ttgagtggtg agaaggaagg tctccttcag ctaccttctg acaaggctct tttgtctgac
     4141 cctgtattcc gccctctcgt tgacaaatat gcagcggacg aagatgcctt ctttgctgat
     4201 tacgctgagg ctcaccaaaa gctttccgag cttgggtttg ctgatgccgg cggatccggg
     4261 ggaagcgact acaaggacca cgacggtgac tacaaggacc acgacatcga ctacaaggac
     4321 gacgacgaca agtaatgagt cgacattaat taacgccccc cccccctaac gttactggcc
     4381 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     4441 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     4501 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagtag
     4561 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     4621 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     4681 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     4741 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     4801 tgggatctga tctggggcct cggtgcacat gctttacatg tatttagtcg aggttaaaaa
     4861 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     4921 atggccacaa ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     4981 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     5041 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     5101 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     5161 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     5221 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     5281 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     5341 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     5401 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     5461 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     5521 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     5581 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     5641 ctgtacaagt aaagcggccg cgtcgacaat caacctctgg attacaaaat ttgtgaaaga
     5701 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     5761 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     5821 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     5881 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     5941 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     6001 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     6061 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     6121 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     6181 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     6241 tgggccgcct ccccgcctgg aattcgagct cggtaccttt aagaccaatg acttacaagg
     6301 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     6361 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     6421 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     6481 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     6541 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     6601 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     6661 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6721 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     6781 tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     6841 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     6901 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat
     6961 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
     7021 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     7081 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     7141 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     7201 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     7261 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     7321 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     7381 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     7441 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     7501 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     7561 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     7621 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     7681 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     7741 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     7801 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     7861 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     7921 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     7981 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     8041 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     8101 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     8161 gaccgaagga gctaaccgct tttttgcaca acatgggaga tcatgtaact cgccttgatc
     8221 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     8281 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     8341 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     8401 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     8461 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     8521 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     8581 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     8641 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     8701 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     8761 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     8821 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     8881 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     8941 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     9001 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     9061 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     9121 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     9181 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     9241 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     9301 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     9361 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     9421 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     9481 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     9541 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     9601 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     9661 ctcattaggc accccaggct ttccacttta tgcttccggc tcgtatgttg tgtggaattg
     9721 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa
     9781 ttaaccctca ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat
     9841 actcttgtag tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa
     9901 aaagcaccgt gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag
     9961 gcaacagacg ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat
    10021 tgtatttaag tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc
    10081 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
    10141 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
    10201 acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg
    10261 aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca
    10321 agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag
    10381 gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa
    10441 aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca
    10501 agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt
    10561 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca
    10621 ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc
    10681 aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa
    10741 gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
    10801 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
    10861 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
    10921 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
    10981 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
    11041 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
    11101 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
    11161 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
    11221 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
    11281 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
    11341 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
    11401 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
    11461 tatagtgaat a
//
