LOCUS       sequence-432003-       11858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Proximity biotinylation of msNLRP3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11858)
  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 11858)
  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(WT)-APEX2-3xFLAG-IRES-eGFP
FEATURES             Location/Qualifiers
     source          1..11858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    116..232
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        235..633
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     regulatory      3841..3850
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3847..4596
                     /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             4615..4680
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    4717..5269
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4861..4878)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5088..5107
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5281..5283,5284..5286,5287..6000)
                     /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(5281..5283,5284..5286,5287..5997)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5587..5606)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5934..5955
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6016..6604
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6069..6089)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6487..6498)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6691..6924
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6996..7117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7033..7052)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7087..7106
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7157..7292
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7312..7331)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7313..7331)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7341..7358)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7341..7357)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7350..7372)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7499..7954
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7586..7605)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7796..7817
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7980..8084
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8085..8153,8154..8945)
                     /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(8303..8322)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9116..9704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9605..9624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9858..9875
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9992..10013
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10028..10033,10034..10051,10052..10058)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10066..10082
                     /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     10071..10093
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10090..10106
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10090..10106
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10125..10145
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10127..10145
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        10173..10399
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             10400..10580
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    10627..10752
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11245..11478
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11663..11707
                     /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 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
       61 gagacagaga cagatccatt cgattagtga acggatctcg acggtatcgg ttaactttta
      121 aaagaaaagg ggggattggg gggtatagtg caggggaaag aatagtagac ataatagcaa
      181 cagacataca aactaaagaa ttacaaaaac aaattacaaa attcaaaatt ttatggcctc
      241 cgcgccgggt tttggcgcct cccgcgggcg cccccctcct cacggcgagc gctgccacgt
      301 cagacgaagg gcgcagcgag cgtcctgatc cttccgcccg gacgctcagg acagcggccc
      361 gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg
      421 acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt
      481 agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat
      541 ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt
      601 tcttgtttgt ggatcgctgt gatcgtcact tggggagacg ccatccacgc tgttttgacc
      661 tccatagaag acaccgacgc caccatgacg agtgtccgtt gcaagctggc tcagtatcta
      721 gaggaccttg aagatgtgga cctcaagaaa ttcaaaatgc atttggaaga ttacccgccc
      781 gagaaaggct gtatcccagt ccccaggggc cagatggaga aggcagatca cttggatcta
      841 gccacactca tgattgactt caatggcgag gagaaggcct gggccatggc tgtgtggatc
      901 tttgctgcga tcaacaggcg agacctctgg gaaaaagcta agaaggacca gccagagtgg
      961 aatgacacgt gtacatcaca ttcctctatg gtatgccagg aggacagcct tgaagaagag
     1021 tggatgggtt tgctgggata tctctcccgc atctccattt gtaaaaagaa gaaagattac
     1081 tgtaagatgt acagacgaca tgtgagaagc aggttctact ctatcaagga caggaacgcg
     1141 cgtctaggtg agagtgtgga cctcaacagt cgctacacgc agctccaact ggtcaaggag
     1201 catccaagca agcaggagcg ggagcatgaa ctcctgacca tcggccggac taaaatgcgg
     1261 gacagcccca tgagttccct taagctggag ctgctgtttg agcccgagga cgggcactcg
     1321 gagcctgtgc acacagtggt gttccaggga gcagcaggca tcgggaaaac catcctagcc
     1381 aggaagatta tgttggactg ggcactggga aagctcttca aagacaaatt tgactatttg
     1441 ttctttatcc actgccgaga ggtgagcctc aggacgccaa ggagtctagc agacctgatt
     1501 gtcagctgct ggcctgaccc aaacccacca gtgtgcaaga tcctgcgcaa gccttccagg
     1561 atcctcttcc tcatggatgg ctttgatgag ctacaagggg cctttgacga gcacattggg
     1621 gaggtctgca cagactggca aaaggctgtg cggggagaca ttctgctaag cagcctcatc
     1681 cgaaagaaac tgctgcccaa ggcctctctg ctcataacga cgaggccggt agccttggag
     1741 aaactgcagc atctcctgga ccacccccgc catgtggaga tcctaggttt ctctgaggcc
     1801 aaaaggaagg agtatttctt taagtatttc tccaacgagc tgcaggcccg ggaggccttc
     1861 aggctgatcc aagagaatga ggtcctcttt accatgtgct tcatccccct ggtctgctgg
     1921 attgtgtgca cggggctaaa gcaacagatg gagaccggga agagcctggc ccagacctcc
     1981 aagaccacta cggccgtcta cgtcttcttc ctttccagcc tgctgcaatc ccgggggggc
     2041 attgaggagc atctcttctc tgactaccta caggggctct gttcactggc tgcggatgga
     2101 atttggaacc agaaaatcct atttgaggag tgtgatctgc ggaagcacgg cctgcagaag
     2161 actgacgtct ccgctttcct gaggatgaac gtgttccaga aggaagtgga ctgcgagaga
     2221 ttctacagct tcagccacat gactttccag gagttcttcg ctgctatgta ctatttgctg
     2281 gaagaggagg cagaggggga gaccgtgagg aaaggaccag gaggttgttc agatcttctg
     2341 aaccgagacg tgaaggtcct actagaaaat tacggcaagt ttgaaaaagg ctatctgatt
     2401 tttgttgtcc gattcctctt tggccttgta aaccaggaga gaacctctta tttggagaag
     2461 aaactaagtt gcaagatctc tcagcaagtc agactggaac tactgaagtg gattgaagtg
     2521 aaagccaagg ccaagaagct gcagtggcag cccagccaac tggaactgtt ctactgcctg
     2581 tacgagatgc aggaggaaga ctttgtgcag agtgccatgg accactttcc caaaattgag
     2641 atcaacctct ctaccagaat ggaccacgtg gtttcctcct tttgtattaa gaactgtcat
     2701 agggtcaaaa cgctttccct gggttttttt cacaactcgc ccaaggagga agaagaagag
     2761 aggagaggag gtcgaccctt ggaccaggtt cagtgtgttt tcccagacac tcatgttgcc
     2821 tgttcttcca gactggtgaa ctgctgcctc acttctagct tctgccgtgg tctcttctca
     2881 agtctaagca ccaaccggag cctcactgaa ctggacctca gtgacaatac tctgggagac
     2941 ccgggcatga gggtgctgtg tgaggcactc cagcacccag gctgtaacat tcagagactg
     3001 tggttggggc gctgcggact gtcccatcaa tgctgcttcg acatctcctc tgtcctgagc
     3061 agcagccaga agctggtgga gctggacctc agtgacaatg ccctggggga ctttggaatc
     3121 agattgctgt gtgtgggact gaagcacctg ctctgcaacc tccagaaact gtggttggtg
     3181 agctgctgtc tcacatccgc gtgttgtcag gatctcgcat tggttctgag ctccaaccat
     3241 tctctgacca gactgtacat tggagaaaat gccttgggag actcaggagt ccaagttttg
     3301 tgtgaaaaga tgaaggaccc acagtgtaac ttgcagaagc tggggttggt gaattccggc
     3361 cttacttcaa tctgttgttc agctctgacc tctgtgctca aaaccaacca gaacttcaca
     3421 cacctctatc tacgaagcaa tgcccttgga gacacaggac tcaggctcct ctgtgagggg
     3481 cttctgcacc cggactgtaa actacagatg ctggaattag acaactgcag cctcacctca
     3541 cacagctgct ggaatctctc cacaattctg acccacaacc acagccttcg gaagctgaac
     3601 ctgggcaaca atgatcttgg cgatctgtgc gtggtgaccc tctgtgaggt gctgaaacag
     3661 cagggctgcc tcctgcagag cctacagttg ggtgaaatgt acttaaatcg tgaaacaaaa
     3721 cgtgccttag aagcgctcca ggaagaaaag cctgagctga ctatagtctt cgagatttcc
     3781 tggggcggat ccgggggaag cggcggatcc atcgaagacg gaagagcgga tccaccggtc
     3841 gccaccatgg gaaagtctta cccaactgtg agtgctgatt accaggacgc cgttgagaag
     3901 gcgaagaaga agctcagagg cttcatcgct gagaagagat gcgctcctct aatgctccgt
     3961 ttggcattcc actctgctgg aacctttgac aagggcacga agaccggtgg acccttcgga
     4021 accatcaagc accctgccga actggctcac agcgctaaca acggtcttga catcgctgtt
     4081 aggcttttgg agccactcaa ggcggagttc cctattttga gctacgccga tttctaccag
     4141 ttggctggcg ttgttgccgt tgaggtcacg ggtggaccta aggttccatt ccaccctgga
     4201 agagaggaca agcctgagcc accaccagag ggtcgcttgc ccgatcccac taagggttct
     4261 gaccatttga gagatgtgtt tggcaaagct atggggctta ctgaccaaga tatcgttgct
     4321 ctatctgggg gtcacactat tggagctgca cacaaggagc gttctggatt tgagggtccc
     4381 tggacctcta atcctcttat tttcgacaac tcatacttca cggagttgtt gagtggtgag
     4441 aaggaaggtc tccttcagct accttctgac aaggctcttt tgtctgaccc tgtattccgc
     4501 cctctcgttg acaaatatgc agcggacgaa gatgccttct ttgctgatta cgctgaggct
     4561 caccaaaagc tttccgagct tgggtttgct gatgccggcg gatccggggg aagcgactac
     4621 aaggaccacg acggtgacta caaggaccac gacatcgact acaaggacga cgacgacaag
     4681 taatgagtcg acattaatta acgccccccc cccctaacgt tactggccga agccgcttgg
     4741 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     4801 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     4861 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagtagtt cctctggaag
     4921 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     4981 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     5041 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     5101 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     5161 tggggcctcg gtgcacatgc tttacatgta tttagtcgag gttaaaaaaa cgtctaggcc
     5221 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc
     5281 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     5341 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     5401 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     5461 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     5521 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     5581 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     5641 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     5701 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     5761 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     5821 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     5881 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     5941 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     6001 agcggccgcg tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     6061 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     6121 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     6181 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     6241 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     6301 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     6361 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     6421 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     6481 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     6541 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     6601 ccgcctggaa ttcgagctcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     6661 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
     6721 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     6781 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     6841 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     6901 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     6961 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     7021 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     7081 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     7141 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     7201 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     7261 ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt
     7321 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     7381 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     7441 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     7501 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     7561 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     7621 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     7681 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     7741 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     7801 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     7861 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     7921 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     7981 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     8041 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     8101 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     8161 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     8221 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     8281 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     8341 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     8401 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     8461 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     8521 taaccgcttt tttgcacaac atgggagatc atgtaactcg ccttgatcgt tgggaaccgg
     8581 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     8641 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     8701 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     8761 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     8821 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     8881 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     8941 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     9001 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     9061 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     9121 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     9181 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     9241 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     9301 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     9361 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     9421 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     9481 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     9541 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     9601 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     9661 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     9721 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     9781 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     9841 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     9901 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     9961 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
    10021 cccaggcttt ccactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
    10081 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
    10141 aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc
    10201 ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc
    10261 atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg
    10321 tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg
    10381 cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc
    10441 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
    10501 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
    10561 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
    10621 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
    10681 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
    10741 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
    10801 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
    10861 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
    10921 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
    10981 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
    11041 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat
    11101 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
    11161 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
    11221 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
    11281 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
    11341 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
    11401 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
    11461 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
    11521 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
    11581 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
    11641 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
    11701 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
    11761 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
    11821 agttaggcag ggatattcac cattatcgtt tcagaccc
//
