LOCUS       sequence-431947-       11858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Proximity biotinylation of msNLRP3(LRRm2, 
            N1008R_R1009E_E1010R_R1013E).
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(LRRm2)-APEX2-3xFLAG-IRES-eGFP
            ...
FEATURES             Location/Qualifiers
     source          1..11858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1058..1284
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1285..1465
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1512..1637
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2130..2363
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2548..2592
                     /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    2859..2975
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2978..3376
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     regulatory      6584..6593
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6590..7339
                     /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             7358..7423
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    7460..8012
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7604..7621)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7831..7850
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(8024..8026,8027..8029,8030..8743)
                     /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(8024..8026,8027..8029,8030..8740)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8330..8349)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8677..8698
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    8759..9347
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8812..8832)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9230..9241)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9434..9667
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9739..9860
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9776..9795)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9830..9849
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9900..10035
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10055..10074)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10056..10074)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10084..10101)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10084..10100)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10093..10115)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10242..10697
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10329..10348)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10539..10560
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10723..10827
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10828..10896,10897..11688)
                     /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(11046..11065)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttccact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg
     1081 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac
     1141 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag
     1201 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt
     1261 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag
     1321 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     1381 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     1441 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     1501 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     1561 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     1621 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     1681 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     1741 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     1801 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     1861 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     1921 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     1981 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     2041 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     2101 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     2161 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     2221 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     2281 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     2341 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     2401 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     2461 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     2521 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     2581 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     2641 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     2701 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     2761 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc
     2821 attcgattag tgaacggatc tcgacggtat cggttaactt ttaaaagaaa aggggggatt
     2881 ggggggtata gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     2941 gaattacaaa aacaaattac aaaattcaaa attttatggc ctccgcgccg ggttttggcg
     3001 cctcccgcgg gcgcccccct cctcacggcg agcgctgcca cgtcagacga agggcgcagc
     3061 gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc ataagactcg
     3121 gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg tgactctagg
     3181 gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct tctcggcgat
     3241 tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga cgcgccgggt
     3301 gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc
     3361 tgtgatcgtc acttggggag acgccatcca cgctgttttg acctccatag aagacaccga
     3421 cgctaccatg acgagtgtcc gttgcaagct ggctcagtat ctagaggacc ttgaagatgt
     3481 ggacctcaag aaattcaaaa tgcatttgga agattacccg cccgagaaag gctgtatccc
     3541 agtccccagg ggccagatgg agaaggcaga tcacttggat ctagccacac tcatgattga
     3601 cttcaatggc gaggagaagg cctgggccat ggctgtgtgg atctttgctg cgatcaacag
     3661 gcgagacctc tgggaaaaag ctaagaagga ccagccagag tggaatgaca cgtgtacatc
     3721 acattcctct atggtatgcc aggaggacag ccttgaagaa gagtggatgg gtttgctggg
     3781 atatctctcc cgcatctcca tttgtaaaaa gaagaaagat tactgtaaga tgtacagacg
     3841 acatgtgaga agcaggttct actctatcaa ggacaggaac gcgcgtctag gtgagagtgt
     3901 ggacctcaac agtcgctaca cgcagctcca actggtcaag gagcatccaa gcaagcagga
     3961 gcgggagcat gaactcctga ccatcggccg gactaaaatg cgggacagcc ccatgagttc
     4021 ccttaagctg gagctgctgt ttgagcccga ggacgggcac tcggagcctg tgcacacagt
     4081 ggtgttccag ggagcagcag gcatcgggaa aaccatccta gccaggaaga ttatgttgga
     4141 ctgggcactg ggaaagctct tcaaagacaa atttgactat ttgttcttta tccactgccg
     4201 agaggtgagc ctcaggacgc caaggagtct agcagacctg attgtcagct gctggcctga
     4261 cccaaaccca ccagtgtgca agatcctgcg caagccttcc aggatcctct tcctcatgga
     4321 tggctttgat gagctacaag gggcctttga cgagcacatt ggggaggtct gcacagactg
     4381 gcaaaaggct gtgcggggag acattctgct aagcagcctc atccgaaaga aactgctgcc
     4441 caaggcctct ctgctcataa cgacgaggcc ggtagccttg gagaaactgc agcatctcct
     4501 ggaccacccc cgccatgtgg agatcctagg tttctctgag gccaaaagga aggagtattt
     4561 ctttaagtat ttctccaacg agctgcaggc ccgggaggcc ttcaggctga tccaagagaa
     4621 tgaggtcctc tttaccatgt gcttcatccc cctggtctgc tggattgtgt gcacggggct
     4681 aaagcaacag atggagaccg ggaagagcct ggcccagacc tccaagacca ctacggccgt
     4741 ctacgtcttc ttcctttcca gcctgctgca atcccggggg ggcattgagg agcatctctt
     4801 ctctgactac ctacaggggc tctgttcact ggctgcggat ggaatttgga accagaaaat
     4861 cctatttgag gagtgtgatc tgcggaagca cggcctgcag aagactgacg tctccgcttt
     4921 cctgaggatg aacgtgttcc agaaggaagt ggactgcgag agattctaca gcttcagcca
     4981 catgactttc caggagttct tcgctgctat gtactatttg ctggaagagg aggcagaggg
     5041 ggagaccgtg aggaaaggac caggaggttg ttcagatctt ctgaaccgag acgtgaaggt
     5101 cctactagaa aattacggca agtttgaaaa aggctatctg atttttgttg tccgattcct
     5161 ctttggcctt gtaaaccagg agagaacctc ttatttggag aagaaactaa gttgcaagat
     5221 ctctcagcaa gtcagactgg aactactgaa gtggattgaa gtgaaagcca aggccaagaa
     5281 gctgcagtgg cagcccagcc aactggaact gttctactgc ctgtacgaga tgcaggagga
     5341 agactttgtg cagagtgcca tggaccactt tcccaaaatt gagatcaacc tctctaccag
     5401 aatggaccac gtggtttcct ccttttgtat taagaactgt catagggtca aaacgctttc
     5461 cctgggtttt tttcacaact cgcccaagga ggaagaagaa gagaggagag gaggtcgacc
     5521 cttggaccag gttcagtgtg ttttcccaga cactcatgtt gcctgttctt ccagactggt
     5581 gaactgctgc ctcacttcta gcttctgccg tggtctcttc tcaagtctaa gcaccaaccg
     5641 gagcctcact gaactggacc tcagtgacaa tactctggga gacccgggca tgagggtgct
     5701 gtgtgaggca ctccagcacc caggctgtaa cattcagaga ctgtggttgg ggcgctgcgg
     5761 actgtcccat caatgctgct tcgacatctc ctctgtcctg agcagcagcc agaagctggt
     5821 ggagctggac ctcagtgaca atgccctggg ggactttgga atcagattgc tgtgtgtggg
     5881 actgaagcac ctgctctgca acctccagaa actgtggttg gtgagctgct gtctcacatc
     5941 cgcgtgttgt caggatctcg cattggttct gagctccaac cattctctga ccagactgta
     6001 cattggagaa aatgccttgg gagactcagg agtccaagtt ttgtgtgaaa agatgaagga
     6061 cccacagtgt aacttgcaga agctggggtt ggtgaattcc ggccttactt caatctgttg
     6121 ttcagctctg acctctgtgc tcaaaaccaa ccagaacttc acacacctct atctacgaag
     6181 caatgccctt ggagacacag gactcaggct cctctgtgag gggcttctgc acccggactg
     6241 taaactacag atgctggaat tagacaactg cagcctcacc tcacacagct gctggaatct
     6301 ctccacaatt ctgacccaca accacagcct tcggaagctg aacctgggca acaatgatct
     6361 tggcgatctg tgcgtggtga ccctctgtga ggtgctgaaa cagcagggct gcctcctgca
     6421 gagcctacag ttgggtgaaa tgtacttaag agaaagaaca aaagaagcct tagaagcgct
     6481 ccaggaagaa aagcctgagc tgactatagt cttcgagatt tcctggggcg gatccggggg
     6541 aagcggcgga tccatcgaag acggaagagc ggatccaccg gtcgccacca tgggaaagtc
     6601 ttacccaact gtgagtgctg attaccagga cgccgttgag aaggcgaaga agaagctcag
     6661 aggcttcatc gctgagaaga gatgcgctcc tctaatgctc cgtttggcat tccactctgc
     6721 tggaaccttt gacaagggca cgaagaccgg tggacccttc ggaaccatca agcaccctgc
     6781 cgaactggct cacagcgcta acaacggtct tgacatcgct gttaggcttt tggagccact
     6841 caaggcggag ttccctattt tgagctacgc cgatttctac cagttggctg gcgttgttgc
     6901 cgttgaggtc acgggtggac ctaaggttcc attccaccct ggaagagagg acaagcctga
     6961 gccaccacca gagggtcgct tgcccgatcc cactaagggt tctgaccatt tgagagatgt
     7021 gtttggcaaa gctatggggc ttactgacca agatatcgtt gctctatctg ggggtcacac
     7081 tattggagct gcacacaagg agcgttctgg atttgagggt ccctggacct ctaatcctct
     7141 tattttcgac aactcatact tcacggagtt gttgagtggt gagaaggaag gtctccttca
     7201 gctaccttct gacaaggctc ttttgtctga ccctgtattc cgccctctcg ttgacaaata
     7261 tgcagcggac gaagatgcct tctttgctga ttacgctgag gctcaccaaa agctttccga
     7321 gcttgggttt gctgatgccg gcggatccgg gggaagcgac tacaaggacc acgacggtga
     7381 ctacaaggac cacgacatcg actacaagga cgacgacgac aagtaatgag tcgacattaa
     7441 ttaacgcccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     7501 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     7561 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     7621 caaggtctgt tgaatgtcgt gaaggaagta gttcctctgg aagcttcttg aagacaaaca
     7681 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     7741 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     7801 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     7861 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     7921 tgctttacat gtatttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     7981 gtggttttcc tttgaaaaac acgatgataa tatggccaca accatggtga gcaagggcga
     8041 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     8101 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     8161 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     8221 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     8281 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     8341 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     8401 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     8461 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     8521 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     8581 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     8641 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     8701 cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc gcgtcgacaa
     8761 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     8821 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     8881 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     8941 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     9001 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     9061 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     9121 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     9181 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     9241 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     9301 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattcgagc
     9361 tcggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     9421 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
     9481 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     9541 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     9601 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     9661 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     9721 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     9781 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     9841 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
     9901 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     9961 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
    10021 gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg
    10081 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
    10141 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
    10201 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
    10261 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
    10321 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
    10381 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
    10441 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
    10501 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
    10561 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
    10621 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
    10681 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
    10741 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
    10801 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
    10861 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
    10921 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
    10981 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
    11041 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
    11101 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
    11161 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
    11221 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
    11281 aacatgggag atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
    11341 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
    11401 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
    11461 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
    11521 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
    11581 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
    11641 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
    11701 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
    11761 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
    11821 tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
