LOCUS       sequence-400583-       13268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid for CRISPR/Cas9-mediated insertion of INSPECT into 
            human NEAT1 locus (both isoforms). Encodes for intronic IRES-driven 
            SigP:NLuc. Plasmid contains FRT-site flanked PuroR-TK cassette..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13268)
  AUTHORS   Truong DJ, Armbrust N, Geilenkeuser J, Lederer EM, Santl TH, Beyer 
            M, Ittermann S, Steinmassl E, Dyka M, Raffl G, Phlairaharn T, 
            Greisle T, Zivanic M, Grosch M, Drukker M, Westmeyer GG
  TITLE     Intron-encoded cistronic transcripts for minimally invasive 
            monitoring of coding and non-coding RNAs.
  JOURNAL   Nat Cell Biol. 2022 Nov;24(11):1666-1676. doi: 
            10.1038/s41556-022-00998-6. Epub 2022 Nov 7.
  PUBMED    36344775
REFERENCE   2  (bases 1 to 13268)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41556-022-00998-6"; journalName: "Nat Cell Biol"; date: 
            "2022-11"; volume: "24"; issue: "11"; pages: "1666-1676"
COMMENT     Sequence Label: pINSPECT:NEAT1-total_SigP:NLuc
FEATURES             Location/Qualifiers
     source          1..13268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2141..2174
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2245..2278
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     polyA_signal    2877..2998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2914..2933)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2968..2987
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3030..3049
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    3048..3103
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     protein_bind    complement(3210..3243)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3314..3347)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             complement(3396..3452)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(4587..4643)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     5230..5249
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(5276..6458)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5285..6227
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(5308..5328)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    join(6534..7107,7108..7110,7111..7119)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6701..6718)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6928..6947
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(11080..11100)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(11080..11098)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11119..11135)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11119..11135)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11132..11154)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11143..11159
                     /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)."
     promoter        complement(join(11167..11173,11174..11191,11192..11197))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11212..11233
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11350..11367)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11521..12109)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11601..11620)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12280..13071,13072..13140))
                     /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     12903..12922
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13141..13245)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgcgcg cctgggtgta gttgtggggg aggaagtggc tagctcaggg cttcagggga
      601 cagacaggga gagatgactg agttagatga gactaggggg cgggctgggg gtgcgagaag
      661 gaagcttggc aaggagacta ggtctagggg gaccacagtg gggcaggctg catggaaaat
      721 atccgcaggg tcccccaggc agaacagcca cgctccaggc caggctgtcc ctactgcctg
      781 gtggaggggg aacttgacct ctgggagggc gccgctcttg catagctgag cgagcccggg
      841 tgcgctggtc tgtgtggaag gaggaaggca gggagaggta gaaggggtgg aggagtcagg
      901 aggaataggc cgcagcagcc ctggaaatga tcaggaaggc aggcagtggg tgcagggctg
      961 caggagggcc gggagggcta atcttcaact tgtccatgcc agcagcccct ttttttccag
     1021 accaagggct gtgaacccgc ctggggatga ggcctggtct tgtggaactg aacttagctc
     1081 gacggggctg accgctctgg cccagggtgg tatgtaattt tcgctcggcc tgggacgggg
     1141 cccaggccgg gcccagcctg gtggagcgtc caggtctggg tgcgaagcca ggcccctggg
     1201 cggaggtgag gggtggtctg aggagtgatg tggagttaag gcgccatcct caccggtgac
     1261 tggtgcggca cctagcatgt ttgacaggcg gggactgcga ggcacgctgc tcgggtgttg
     1321 gggacaacat tgaccaacgc tttattttcc aggtggcagt gctccttttg gacttttctc
     1381 taggtttggc gctaaactct tcttgtgagc tcactccacc ccttcttcct ccctttaact
     1441 tatccattca cttaaaacat tagctggtca tctggtaagc ccgggacagt aagccgagtg
     1501 gctgttggag tcggtattgt tggtaatggt ggaggaagag aggccttccc gctgaggctg
     1561 gggtggggcg gatcggtgtt gcttgcctgc agagagggtg gggagtgaat gtgcaccctt
     1621 gggtgggcct gcagccatcc agctgaaagt tacaaaaatg cttcatggac cgtggtttgt
     1681 tactatagtg ttcctcatgg cgagcagatg gaaccgggag acatggagtc cctggccagt
     1741 gtgagtccta gcattgcagg aggggagacc ctggaggaga gagcccgcct caattgatgc
     1801 ctgcagattg aatttccaga ggcttaggag gaggaagttc tccaatgttc tgtttccagg
     1861 ccttgctcag gaagccctgt attcaggagg ctaccattta aagtttgcag atgagcttat
     1921 ggggggcaat cttaaaaagt ccacagcaga tgcatccggc tcgaggggcc atcagctttg
     1981 aataaatgct tgttccagag cccatgaatg ccagcaggca cccctccttt cctggggtaa
     2041 aggtaagtat cctttttaca gcacaactta atgagacaga tagaaactgg tcttgtagaa
     2101 acagagtaga ttttatcgaa ttgcaacctc ttcgagggac ataacttcgt ataatgtatg
     2161 ctatacgaag ttatgggtcg atgctgagat ctggactaga gggtcgatgg tgatgcttgg
     2221 atcaaaatag gaagaatccc aatgataact tcgtataaag tatcctatac gaagttatac
     2281 tagaattgcg taccgccacc atgtcgccaa ggccacctcc cctgtgagct agactggaca
     2341 gccaatgacg ggtaagagag tgacatttct cactaaccta agacaggagg gccgtcaaag
     2401 ctactgccta atccaatgac gggtaatagt gacaagaaat gtatcactcc aacctaagac
     2461 aggcgcagcc tccgagggat gtgtcttttg ttttttataa ttaaaaaggg tgacacgtga
     2521 ggccacctcc cctgtgagct agactggaca gccaatgacg ggtaagagag tgacatttct
     2581 cactaaccta agacaggagg gccgtcaaag ctactgccta atccaatgac gggtaatagt
     2641 gacaagaaat gtatcactcc aacctaagac aggcgcagcc tccgagggat gtgtcttttg
     2701 ttttttataa ttaaaaaggg tgaagccgac acacaaaaaa ccaacacaca gatctctaga
     2761 gctctggatc ttttattgcc cgggctacca catttgtaga ggttttactt gctttaaaaa
     2821 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     2881 tgtttattgc agcttataat ggttacaaat atagcaatag catcacaaat ttcacaaata
     2941 tagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3001 atgtctgtat actttaaatg atttgccctc ccatatgtcc ttccgagtga gagacacaaa
     3061 aaattccaac acactattgc aatgaaaata catttccttt attagccaga agtcagatgc
     3121 cagcacaaca accagcacgt tgcccaggag ctgtaggaaa aagaagaagg catgaacata
     3181 gttagcagag ttacgtagcg atcgctagta taacttcgta tagcatacat tatacgaagt
     3241 tatgggtcga tggtgacaat agcgaaaaag aaagaacaat caagggtccc caaactcacc
     3301 taggtccttt gatataactt cgtataggat actttatacg aagttatggc atctccttcg
     3361 gaagttccta ttctatttga agtataggaa cttccaggtc caggattttc ctcgacgtcc
     3421 cctgcctgct tgagcagtga aaagttagtt gccccggatc cgttggcttc gcccatctct
     3481 ctagcgaagg ttctggcgag gtcacagatg gtggggatgc tgccaggggt tgtcacatgg
     3541 gtctgcacca tgccgcttgt cagttgcagc agggcatctc tgcatccggc tgggctctga
     3601 tcgtagtcca ggatgaacac atgcatgggc cgcagtcttt ttgccagcac gtcgagagcc
     3661 caggcgaaca cgttatacag gtcgccgtta ggggccagca gttcaggggc tctgaacagt
     3721 gtgaacaggg tgtcgccgat gtgaggtctg ggtccagcgt tagactgagg ctcagcgcct
     3781 tgtggaggaa ctgctgtgcc agagagctgt ccccagtcct ctctccagga accgccgcct
     3841 tgaagatagc gcacggtgtt agccagcagg ccgtacactc ttctgatggc ggccagcata
     3901 gccagatcca gtctctcgcc aggcctctgt ctcttagcca gtctgtcgat gtgtctgtcc
     3961 tctggcaggg ctccaagcac gatgtttgtg ccgggcagtg taggaggaat cagagccacg
     4021 aaagccagca cggcctgagg tgtcatgctg cccatcagat accgagcggc agggtaacac
     4081 agcagggcga agatggggtg tctgtcgaag atcagggtca gagcaggtgg tggagcgtga
     4141 gaagatccag cctcgccgcc aatatgtgga gccagcacag cgtctgtcac ggcgtaaggc
     4201 atgcccattg tgatctgggc agatgtcatg accacagcag cgtcgcctgc gctaatttcg
     4261 ccctggtcca gtctgtgctg tgtggtgtag atgttggcga ttgtctcaga ggctcccagg
     4321 acctgccagt aggtcatagg ctcgggcacg tacacgatgt catctctgct gcccagagcc
     4381 accagcagct gtgttgttgt ggtcttgccc atgccatgag ggccgtcgat atacactctc
     4441 agcagtgtgg gcatcttctg ttccagccgc acttctgtag cctcttgctg ccttcttggc
     4501 ctgagggctg ttcttctgtt gctgtggcct ctgcttctag cggcttgatc gaaggcgctg
     4561 gcgtgctggt gacaaggata agaagcagga ccggggttct cttccacgtc gccagcctgt
     4621 ttcagcaggc tgaagtttgt agcgccggat ccggcgccag gctttctggt catgcaccag
     4681 gttctagggc cctcaggcac ttccacgtcg gcggtcacgg tgaagcccag tctctcgtag
     4741 aagggcaggt ttctgggggc gcttgtttcc aggaaggcgg gcacgccagc cctttcagca
     4801 gcttccaccc caggcagcac cacagcagat cccagtccct tgccctggtg gtcaggtgac
     4861 acgcccacgg tggccagaaa ccaggcaggc tcttttggtc tgtggggggc cagcaggcct
     4921 tccatctgct gctgggcagc cagtctagag ccgctcagct cggccattct aggtccgatc
     4981 tcggcgaaca cagcgccggc ttccacagac tcaggggttg tccacacagc cacagcggcg
     5041 ccatcatcgg ccacccacac tttgccgatg tccaggccca ctctggtcag aaacagttcc
     5101 tgcagctcgg tcactctctc gatgtgccgg tcggggtcca cggtgtgtct tgtggcaggg
     5161 taatcggcga aggcagcggc cagtgtccgc acagctcttg gcacatcgtc cctggtggcc
     5221 agccgcactg tgggcttgta ctcggtcatg gtggcgcgcc ttttaggggt agttttcacg
     5281 acacctgaaa tggaagaaaa aaactttgaa ccactgtctg aggcttgaga atgaaccaag
     5341 atccaaactc aaaaagggca aattccaagg agaattacat caagtgccaa gctggcctaa
     5401 cttcagtctc cacccactca gtgtggggaa actccatcgc ataaaacccc tccccccaac
     5461 ctaaagacga cgtactccaa aagctcgaga actaatcgag gtgcctggac ggcgcccggt
     5521 actccgtgga gtcacatgaa gcgacggctg aggacggaaa ggcccttttc ctttgtgtgg
     5581 gtgactcacc cgcccgctct cccgagcgcc gcgtcctcca ttttgagctc cctgcagcag
     5641 ggccgggaag cggccatctt tccgctcacg caactggtgc cgaccgggcc agccttgccg
     5701 cccagggcgg ggcgatacac ggcggcgcga ggccaggcac cagagcaggc cggccagctt
     5761 gagactaccc ccgtccgatt ctcggtggcc gcgctcgcag gccccgcctc gccgaacatg
     5821 tgcgctggga cgcacgggcc ccgtcgccgc ccgcggcccc aaaaaccgaa ataccagtgt
     5881 gcagatcttg gcccgcattt acaagactat cttgccagaa aaaaagcgtc gcagcaggtc
     5941 atcaaaaatt ttaaatggct agagacttat cgaaagcagc gagacaggcg cgaaggtgcc
     6001 accagattcg cacgcggcgg ccccagcgcc caggccaggc ctcaactcaa gcacgaggcg
     6061 aaggggctcc ttaagcgcaa ggcctcgaac tctcccaccc acttccaacc cgaagctcgg
     6121 gatcaagaat cacgtactgc agccaggggc gtggaagtaa ttcaaggcac gcaagggcca
     6181 taacccgtaa agaggccagg cccgcgggaa ccacacacgg cacttacctg tgttctggcg
     6241 gcaaacccgt tgcgaaaaag aacgttcacg gcgactactg cacttatata cggttctccc
     6301 ccaccctcgg gaaaaaggcg gagccagtac acgacatcac tttcccagtt taccccgcgc
     6361 caccttctct aggcaccggt tcaattgccg acccctcccc ccaacttctc ggggactgtg
     6421 ggcgatgtgc gctctgccca ctgacgggca ccggagccaa ttcgaagttc ctattctatt
     6481 tgaagtatag gaacttcaaa tacaagggct tactgtgcgg actgggacgc ccccccctct
     6541 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     6601 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     6661 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     6721 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
     6781 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     6841 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     6901 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     6961 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
     7021 tttacatgtg tttagtcgag gttaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     7081 ttttcctttg aaaaacacga tgataatatg gccaccacca tggtggtcaa ggtgctgttc
     7141 gccctgatct gtattgccgt ggccgaggcc aagcctaccg tgttcaccct ggaagatttc
     7201 gtcggcgatt ggagacagac cgccggatac aacctggacc aggtgttgga gcaaggcggc
     7261 gtgtccagcc tgtttcagaa cctgggagtg tccgtgacac ccatccagag aatcgtgctg
     7321 agcggcgaga acggcctgaa gatcgacatc catgtgatca tcccttacga gggcctgtcc
     7381 ggcgatcaga tgggccagat cgagaagatt ttcaaggtgg tgtaccccgt ggacgaccac
     7441 cacttcaaag tgatcctgca ctacggcacc ctggtcatcg atggcgtgac ccctaacatg
     7501 atcgactact tcggcagacc ctacgaggga atcgccgtgt tcgacggcaa gaaaatcacc
     7561 gtgaccggca cactgtggaa cggcaacaag atcatcgacg agcggctgat caaccccgat
     7621 ggcagcctgc tgttcagagt gaccatcaac ggcgtgacag gatggcggct gtgcgaaaga
     7681 atcctggcct aatgtacaga tatcactatc gataccgcgg ggatcctcta gagtagacca
     7741 cctcccctgc gagctaagct ggacagccaa tgacgactaa gagagtgaca tttttcacta
     7801 agctaagaca ggagggccgt cagagctact gcctaatcca aagacggcta aaagtgataa
     7861 aaatgtatca ctccaagtta agacaggcgc agcttccgag ggatttgcct ggccggattg
     7921 actgacgtct taaccactgc atatcgatac cgcggggatc ctctagagta gaccacctcc
     7981 cctgcgagct aagctggaca gccaatgacg actaagagag tgacattttt cactaagcta
     8041 agacaggagg gccgtcagag ctactgccta atccaaagac ggctaaaagt gataaaaatg
     8101 tatcactcca agttaagaca ggcgcagctt ccgagggatt tgcgtttaaa cgcggccgct
     8161 ctatactcga cgattgacta tctgacaaat catagagaac atagagaacc tcctcgagtc
     8221 gcctgctttt ctgcctggta ctaacttctc tcccctctcc tcttttcttt ttctgcaggt
     8281 tttcagatgc tgcatcttct aaattgagcc tccggtcata ctagttttgt gcttggaacc
     8341 ttgcttcaag aagatcccta agctgtagaa cattttaacg ttgatgccac aacgcagatt
     8401 gatgccttgt agatggagct tgcagatgga gccccgtgac ctctcaccta cccacctgtt
     8461 tgcctgcctt cttgtgcgtt tctcggagaa gttcttagcc tgatgaaata acttggggcg
     8521 ttgaagagct gtttaatttt aaatgcctta gactggggat atattagagg aagcagattg
     8581 tcaaattaag ggtgtcattg tgttgtgcta aacgctggga gggtacaagt tggtcattcc
     8641 taaatctgtg tgtgagaaat ggcaggtcta gtttgggcat tgtgattgca ttgcagatta
     8701 ctaggagaag ggaatggtgg gtacaccggt agtgctcttt tgttcttgct tcgttttttt
     8761 aaacttgaac tttacttcgt tagatttcat aatactttct tggcattcta gtaagaggac
     8821 cctgaggtgg gagttgtggg ggacggggag aaggggacag cttggcaccg gtcccgtggg
     8881 cgttgcagtg tgggggatgg gggtatgcag cttggcactg gtactgggag ggatgagggt
     8941 gaagaagggg agagggttgg ttagagatac agtgtgggtg gtgggggtgg taggaaatgc
     9001 aggttgaagg gaattctctg gggctttggg gaatttagtg cgtgggtgag ccaagaaaat
     9061 actaattaat aatagtaagt tgttagtgtt ggttaagttg ttgcttggaa gtgagaagtt
     9121 gcttagaaac tttccaaagt gcttagaact ttaagtgcaa acagacaaac taacaaacaa
     9181 aaattgtttt gctttgctac aaggtgggga agactgaaga agtgttaact gaaaacaggt
     9241 gacacagagt caccagtttt ccgagaacca aagggagggg tgtgtgatgc catctcacag
     9301 gcaggggaaa tgtctttacc agcttcctcc tggtggccaa gacagcctgt ttcagagggt
     9361 tgttttgttt ggggtgtggg tgttatcaag tgaattagtc acttgaaaga tgggcgtcag
     9421 acttgcatac gcagcagatc agcatccttc gctgcccctt agcaacttag gtggttgatt
     9481 tgaaactgtg aaggtgtgat tttttcagga gctggaagtc ttagaaaagc cttgtaaatg
     9541 cctatattgt gggcttttaa cgtatttaag ggaccactta agacgagatt agatgggctc
     9601 ttctggattt gttcctcatt tgtcacaggt gtcttgtgat tgaaaatcat gagcgaagtg
     9661 aaattgcatt gaatttcaag ggaatttagt atgtaaatcg tgccttagaa acacatctgt
     9721 tgtcttttct gtgtttggtc gatattaata atggcaaaat ttttgcctat ctagtatctt
     9781 caaattgtag tctttgtaac aaccaaataa ccttttgtgg tcactgtaaa attaatattt
     9841 ggtagacaga atccatgtac ctttgctaag gttagaatga ataatttatt gtatttttaa
     9901 tttgaatgtt tgtgcttttt aaatgagcca agactagagg ggaaactatc acctaaaatc
     9961 agtttggaaa acaagaccta aaaagggaag gggatgggga ttgtggggag agagtgggcg
    10021 aggtgccttt actacatgtg tgatctgaaa accctgcttg gttctgagct gcgtctattg
    10081 aattggtaaa gtaataccaa tggcttttta tcatttcctt cttcccttta agtttcactt
    10141 gaaattttaa aaatcatggt tatttttatc gttgggatct ttctgtcttc tgggttccat
    10201 tttttaaatg tttaaaaata tgttgacatg gtagttcagt tcttaaccaa tgacttgggg
    10261 atgatgcaaa caattactgt cgttgggatt tagagtgtat tagtcacgca tgtatgggga
    10321 agtagtctcg ggtatgctgt tgtgaaattg aaactgtaaa agtagatggt tgaaagtact
    10381 ggtatgttgc tctgtatggt aagaactaat tctgttacgt catgtacata attactaatc
    10441 acttttcttc ccctttacag cacaaataaa gtttgagttc taaactcatt agaattgttg
    10501 tattgctatg ttacatttct cgacccctat cacattgcct tcataacgac tttggatgta
    10561 tcttcatatt gtagatttag gtctagattt gctagctcca agtaattaag gccatgtagg
    10621 agagcatggt aaccacagat agaactggta ttatcccaag tggtctgcag actgctgagt
    10681 ggggatggga tctgctctct gttgagagtt ggtaatcatt ggtttgaaat gtgatgaaac
    10741 cactcaagcc aatgaaggtg ggtgtgtagg tggggagtac tttgccataa tattttaaaa
    10801 cattacctgg ttagagttct aagtggtact tatttttgtt tggttagggg aaagcctgaa
    10861 taaaaacaga aatggacaca taatatgcat attccatagt ctttgggagg ctggaatgtg
    10921 cctgggattt gggtctaagt gtatgcgtaa ttcttacctc actaaagaat ttgccttgtt
    10981 tttttccttt tggtgagtga ctaaaacgtc tgggcttccc tgtgtgcgtg ctacagtaag
    11041 caagcagagg ctgtgggaga cggtacccag cttttgttcc ctttagtgag ggttaattgc
    11101 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
    11161 tccacacaac atacgagccg ggagcataaa gtgtaaagcc tggggtgcct aatgagtgag
    11221 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
    11281 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
    11341 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
    11401 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
    11461 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
    11521 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
    11581 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
    11641 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
    11701 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
    11761 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
    11821 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
    11881 taacaggatt agcagagcga ggtatgtagg aggtgctaca gagttcttga agtggtggcc
    11941 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
    12001 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
    12061 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
    12121 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
    12181 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
    12241 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
    12301 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
    12361 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
    12421 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
    12481 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
    12541 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
    12601 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
    12661 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
    12721 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
    12781 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
    12841 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
    12901 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
    12961 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
    13021 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
    13081 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
    13141 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
    13201 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
    13261 agtgccac
//
