LOCUS       sequence-484212-       15761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express doxycycline inducible Unspliced CRNDE in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15761)
  AUTHORS   Lee JS, Dan T, Zhang H, Cheng Y, Rehfeld F, Brugarolas J, Mendell JT
  TITLE     An ultraconserved snoRNA-like element in long noncoding RNA CRNDE 
            promotes ribosome biogenesis and cell proliferation.
  JOURNAL   Mol Cell. 2025 Apr 17;85(8):1543-1560.e10. doi: 
            10.1016/j.molcel.2025.03.006. Epub 2025 Apr 3.
  PUBMED    40185099
REFERENCE   2  (bases 1 to 15761)
  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: 
            "10.1016/j.molcel.2025.03.006"; journalName: "Mol Cell"; date: 
            "2025-04-17- 17"; volume: "85"; issue: "8"; pages: "1543-1560.e10"
COMMENT     Sequence Label: XLone-Unspliced CRNDE
FEATURES             Location/Qualifiers
     source          1..15761
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..376
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    17..35
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    53..71
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    89..107
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    125..143
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    161..179
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    197..215
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    233..251
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             5987..5995
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    10723..10857
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   11133..11167
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(11178..11195)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(11178..11194)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11187..11209)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(11403..11422)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11522..11544
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(11582..11600)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        11668..11772
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11773..11841,11842..12633)
                     /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(11991..12010)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12804..13392
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13293..13312
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13546..13563
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13680..13701
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13716..13721,13722..13739,13740..13746)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13754..13770
                     /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     13759..13781
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13778..13794
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13778..13794
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   13820..13882
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    14058..14139
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(14256..14654)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(14655..14711)
                     /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(14721..15464)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             complement(15465..15488)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      15488..15497
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(15517..15728)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
ORIGIN
        1 atcacctcga gtttactccc tatcagtgat agagaacgta tgaagagttt actccctatc
       61 agtgatagag aacgtatgca gactttactc cctatcagtg atagagaacg tataaggagt
      121 ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa
      181 cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc
      241 agtgatagag aacgtataag ctttgcttat gtaaaccagg gcgcctataa aagagtgctg
      301 attttttgag taaacttcaa ttccacaaca cttttgtctt ataccaactt tccgtaccac
      361 ttcctaccct cgtaaaggta ccgcggggag gggcggggtc tcgatcgcgc tattgtcatg
      421 gagacgggaa gctggctgca gcggcggcgg ggaccgtggg gccgaggtgg ctgccagccg
      481 gccaatgtct aagcgaggcg gagcggccca ggcggcccga gcctggggga gcgcgcagcc
      541 ggccagtggc ggcctcgccg gcggcctctt cccgggctcg cagtaggccc gagtcgtcgc
      601 cgggagctcc tgggagcagc gtccccgccc tgctcccctc gctcccgcct cttgcggccc
      661 cacggcccct cagcgcccgc ccccggctcc gcccgccgca gccgcagccc ctggcgctaa
      721 cggtcggtaa cggcccgcgc gcgccgcccg ccgggggctc gcgccagcca cgagggagcg
      781 tccgcggccc gcgcgcccgc gcggcggagg agaggtgagc ccccgcccgg gccaggccct
      841 ctggccgcgc cgtccgcccc tctagtcgtg tcccctcgtg ggccgaacgg acgcggcggt
      901 gccccgcgcc cgaccagacg tcccgtgggc tagggcctgg gcctcgggcc gcgtcggcgc
      961 cggtcgaggt tctccgggtg tcggggttcg gggcgggcgc gcgtgggcgt ggctcctctg
     1021 tccacgcctg ttcccttcgt cgccgcggct ctcgtccggg acacggcttt ccggagtaga
     1081 gcccttggag gtttgtgcca ccgagaaccc gagtctgtca cacagacatc ctcattacat
     1141 catcctgcca ctccggcagt cccgcgcttt ctccccccac ccccgccccc gcccccgccc
     1201 cgtggcttgt ttgttggttg tttttttaat ttttttaacc ccttttcttg tactgtcttc
     1261 tttttggtgt caggggctgg agagactcct gcaagatatt gaggcattta gaaagtatgg
     1321 ttctgttgcc gtggttgtca cctccgctct acgtcaaatt taagactcat ttcttaagga
     1381 tgtctctatt ttctgcttta tttgcacatt gatggctctg ctgctcagct ggccacggcc
     1441 gcccggcgtc tgagatagaa ttgtgttaat atgagaaagg gaacggccga tcatgattag
     1501 caggcagacg gcatgcagat agcatctggc gggttttctc cgtttttata tgaaagcgtt
     1561 cacttgtttg ctggtaaaca cgaggtttta actttttatc tgggtagtgg tgaccgtttg
     1621 tttcactctc ccctcctttt tacttcctat tcattagctt ttccgtttgg aaagctgcat
     1681 acttgttagc ggtagtttgg tgtcttgact ctgaagcttt ctcttccagc agaggtggga
     1741 tctgtatccc tcccatccta ggttaccaag aagacttttt ctccccttac tttctattga
     1801 atttgtgtct ttcccaacat ttaaacttac actagggata agtttgctcg ggttgttttt
     1861 gacgtaggac tattaaatat tctcattttg aaacttccta aaggtttatt ggtttgtcat
     1921 gtctgcaccg atccagtttc gcataactct taaaatttaa agctttaaag tacggttcgc
     1981 aattgtattg tcaagacatg gccctagagc ttgttcttca agttttggct gtacagttgt
     2041 cattgtttca gtcaagtgca aaagaggtta ttcctgttta tttagtgtta atagctattt
     2101 acagttaaaa ctttaaagac tcatagaagt aggtctgctg tattttaaag atctcatcat
     2161 tcttgtggga gagacacaac accaacacca ggaaaggcaa cagattgctt ctgtcatatc
     2221 gctttgctgt tttggagtgt acttaaagga ttatgattga taaggctgtc ttgccagaga
     2281 atgttgacac aggtgtgcat cgttatctta attaagatcc tgagaaaatg ttgtctataa
     2341 atggtgacat aagaagggcg gcatattttt gtacagtttt gcttttggag aatttaattg
     2401 cacttgaaac ctgtctttga gcttccagat tttgtgagca ctttagcaac agaagtttta
     2461 gagagaaatg atagtttaat gttaagtgtt ctggcatgta ctaatagtac ttgtggcttt
     2521 ttggcaatgc ctgtgtcggc tttctttgtg ctctaccttt gtggaataat ggtttctaat
     2581 tttgaaagaa aagcataaaa tagagcagta ttttattaag caaacacttg ttagtgttca
     2641 gaataagtct gtagaggagg gcatgcttaa tgtgcagaaa caggaatata ataagaggta
     2701 aatatatgag aaatttatgc tatttcgttt ttttggaaaa gaaacagtct cctggggtgc
     2761 cttaatgatc taaaacaatt tctttttcat atcattaaat acctagatac aatgtttacg
     2821 gaatgccttg tttttatttc taaaaataga agaaaactgt tttatttctt ccaaaatact
     2881 gttctgtgtg tttgaattga tgagtaagtg aagatagtgc atttagtctc ttactagttt
     2941 aaggatctga gtcataacct gggtgaccac taaaataata ttgctgttct ttactacaaa
     3001 tctcggaatg caaaaacact ggcaagatgg atgctgttga tgatattacc cttggcttac
     3061 ccagcaccta atacctttta acataagtat tgcattaaga ttttctcttc tctttgtagc
     3121 ttaggatttc ctggaagctg gctgtctatc agtgtgtatg taagaagtgt atatgttttt
     3181 ctttaatatt aaaagtcact gtaaaaagag gaaatgaaag tggaggagac cataggcata
     3241 tgatcggaaa agccaggtgt agagtcacag ctcattcttg actccaagga taattacctg
     3301 gttagaatta catggtaaac taaaagaaag acttacgtag atttgtttgc taaaactaaa
     3361 acaagaccat taattttgag ataattgctt tatttatctc ccataccacc cctaatcaaa
     3421 atttttttcc cctctggcag ctatgatgag cattcttatg ctgttatcca tgtgttattt
     3481 tcaaatttga tatattgcat taaaagagat gaaataggaa tatgaactaa ttatccattt
     3541 taacgaataa ctcaataatt attattatgt gtatacattc tagaaatctt aggaaaaaat
     3601 gttctttagt tgataacagg aaagtaaatt aaatcatgta atcctgagtg aattaaatat
     3661 gctcttcaaa atagatgatg aacatagcat catactttgg aataatgttt taactctggg
     3721 atagtacagc ctgttttgag tggtagtatt tcattatctt tatatgtttc tcttcaatat
     3781 ctcttttttg ttgactccta tattttaaca tctttttggt atatttgtta gctgtttata
     3841 gggaataatt tttcttaata gaatagcagc tggtgcaact ctaagaacat gtcaaatcta
     3901 ggaattcttt tttccttttt caataatgag tggtttgctt tcatagcact gctctgaagt
     3961 gtgatgctga aggcttattt ctgcataaat ctgcagtgtt tttatagctg tagtgtacct
     4021 aagcaccaat ttagcttagc aaattaactt agatgcttca aaattgaaaa ccagtatatt
     4081 ctaatgtagt ttgagtttaa gtctttatta gtaaggatgc acttaacatt tgttcaggag
     4141 aagagaatat aattttctcc tgcagttgtc atttacgtaa tccatttttc tagtcttttc
     4201 tcaggctaat gatattccat accttgatac agcttttatt tgtttatttg ccataatatg
     4261 gagtgatacc ttcgagaaaa ttagatattt gaatctaata tcccaaaagt attacttagt
     4321 tttctgtgtt tccttaaaca gaatcatttc cattttcggt gcaggtgtta agtgtgatgc
     4381 ttccataata catttggatg ctgtcagcta agttcacttc tgaactaagg ggttcctcca
     4441 aatgttggct gaaattcatc ccaaggctgg tctgcaagtg agtgtctgca cacagtttgc
     4501 ttgtatgtgg agtcgatcca aaatagcatc aatgttggtt ttaccaaagt atttattatt
     4561 gataatagag gctaagtaca aaatgtagag aatgtcagct acttgaggcc tttgattatt
     4621 aaaaatttta ttaatgcatt aaacaagagt acagtaaata gataaatttt aggttcatga
     4681 aataaaactg aataatttat ttttacttac tatttatcat ggaattactt tgaataattt
     4741 atttttaatg gtataattgg acagtaaaat ttataaactc agtgcttttc ataaaaatca
     4801 aagtgaagtt tgtaatattt tatacaaata gaattattat ttaagagaaa taacctgttt
     4861 atgcctaatt acagttttta atcatttcag ttacctattt cttttaagaa taaatttagt
     4921 gggaatatca gttccagtca tgggtaccaa acttttttag tgacagagta cacacaggta
     4981 tgtaaaactt gtcatttctc atcaaataga ggctgctgaa tataggcagg taagaaaagc
     5041 tatgagaaag aattgttttg cagaatattt gtctagttgt cagagcaagg aactgaattt
     5101 attgacacag aacattttat taagtaaaaa aaatggttct tataacaaaa aaaaaagcta
     5161 attttacaga aggcagtagt taatatagcc gccaataaaa taaatgtttc tctgaatact
     5221 ttccgaggca ttacagtgat ttttaactaa tatgaagtga tatataataa ttttaaagta
     5281 acatccttga gttttcctat tattaattgc ttatggaaat tgggtttcac gtatgactga
     5341 gagctaaagc attacagtga gttagaaaac acaacacaaa tgtaaagaaa atgttaggtg
     5401 gtgagtaatc tgattccttt ttgtttgcct ttaagcttag ttttttgttt tgttttactt
     5461 tgttttaaac catgtataaa attgttgaat ttaaaagata agaggattaa gtaatttctt
     5521 ttttcctcct aaggataaat gtaggaaaaa tctaaacaca taggcagatt ggttgagttt
     5581 tatatctgtt atcggccaca tttattaaga ttcatatttc atgtatatta gaggtattca
     5641 catgtattaa aattcttata ttccttctat ataaaataga tgtagggggt tcccactctt
     5701 gaaaatatga agaaaagatg tcctttcagc aataatgggt tatggttgat taactgagaa
     5761 ggttgtatta aacgttctct agtagaaatg gctaaagagc atgctttttg agaagtgtat
     5821 catctaggaa gaaaatcaaa tggagtattg gtaattaaat tgtaattcca tgaaggaagg
     5881 aagtggtgca aaagatgaag ctaactattc ctgtttttct ttttaagagt ctgcaattca
     5941 taatggagct actgtactgg ctattggaag gaggagattc tgaagataag gaggtaatat
     6001 tatctctttt aaaagaatac tttcctctgt aatcctgaat ctttattaca tgtaagaact
     6061 ttgtgcagta gacagcaatt tctttgaatt tggtatatgg aaacaatttt attttcctct
     6121 gctaagtttt tgagcctgcc tcttctagtg ccatggactg cattggtaga gctgagaaat
     6181 atcatttagc catactcagc acccttaaaa tagcttcttt ctgagaatta gatctgtgaa
     6241 ggtgtcctgc acagttcttg tagatgtcat tttagtttgt ggttgacgtg catgcattta
     6301 gcatgttgct taaccgtcct cattcgcctc ccagttcttt gttgccttca tttgggggga
     6361 tgtgttttct gctggatgat ttactgctag acatgaccaa actctgagta taagacttgg
     6421 tgtttggcag ctggttccag tgctctgctg agaagtagtt gggcccagcc tggggcattg
     6481 tagtgtcctg gaggccggga gctcctgcac aggggttctt ttcctggttc agagccttag
     6541 gtgcctttcc acttttcacg taatcttttc ctaggttggc ttgctgctta ctttgcagct
     6601 gttgcaggga ttcacatgga atggagggct cccttttatt gtggattatt tctttgataa
     6661 tttacccatg tgctttttca ttttcaaaaa cccagtggtg tttaagaatg aaagattctt
     6721 gaagaacaaa aattgggtta aatttgtatc tatcagaaaa gattctatcc tctgatgcta
     6781 tgtaggcaca ttgaatatgc aggctaaatt aaaaacagag taaaaccttt ttataatatg
     6841 ccaattacta tatctaagaa tgtttataca ggctgaaatt ttgtaatggt atttatatct
     6901 gttttccttt ttaaggaaaa taatatactt ttctaggcat caaaaatatc tgcccctttc
     6961 actaatgctg atgtaccacc ttcccccaac ccccaacgct aaatttgact ggcttaaaaa
     7021 catctgcccc ctgaactata tctggggagg aatattaata aaacaatgaa ggacttcatg
     7081 gtgtctagtt atataattag gaaattggtt agaaacgtgg ctaaaaccag tttctttcat
     7141 taaaagaact agtgtacatt taaaaacaaa aaccttcagg aaaaagactc tttgatcttt
     7201 aagtcaaata gtatttttgg ttaacacagc aggaagggga aatataccaa tttcagattc
     7261 ttttatttat gcctaagtag aggttgtaag cagctaagga agtgattaaa atgtagtcta
     7321 gtaaaaaatg gtgctgatta cttggaaagc agtttacatt tgcaaaaaaa ttcagtatta
     7381 tgaccttcac caacttttac acatcatata cttgggttat tacttatggt atagcttgtg
     7441 tatggaatgc aagggtattt tcaaattgac taggtcttgc tggtcatctt aaaatatgtt
     7501 acaatattac aaaattgaaa tgtaatattt tttaatagaa ggaaaatata aatttaatat
     7561 ctgggcaatt gagaccttta aacttacttt aaaagtatga tcttgatgta tatgatactg
     7621 ttttgtcttt gctatattaa cagaattaga ggggtgttct gcaattcaaa taccttatat
     7681 attccaaatt ttattctcta taatggactt ttaaaataaa aggtatatgt gcttcaagag
     7741 ggcaaaattt gaatcatgag ctaatttgct aagcatcaga ttatagaaaa gcatccttga
     7801 ttaatttgga actgtgaaag ggggcgggta aaactgtttt ctgcagaaat ttactagtgc
     7861 agcaaccatt taaattaaat gtttgttaac ataatagtga tggcattttc tcctccccct
     7921 ccttgtggtt ttgtccaact agatgttaca gtggcagttg cactgactgt taagtgttta
     7981 aatgatgaca ccattatgtg aagtgatttt gaaatgagag attccagcca agaattacat
     8041 ctgctcccat ctccttcaaa tcatactctc tggcagtaca gattatgatt gatttgtttg
     8101 tgacagattg caggaaacag tcattgattt ttcaatattt taccttaaaa ttatttacag
     8161 ttgtaaccat ggggaggtat tttcatgggc tgtcagcccc tgaaagacta ggataatatt
     8221 ccctgctctc tgacaagaca aattacctgt aatgagtgca gtagctgaag ggtatacttt
     8281 tattttaaaa tatgtcaata accccagtga ctaaacgaat attgatttag cataatgaag
     8341 cctgagtaac gtgaaaatga gctttttcaa ggggcatggt aaagtctttc tttttagctg
     8401 gttgtaagaa gcttttgatt cttttcagcc agctggtagg aatatagaat tttataagca
     8461 aaccatcagg aatgatagtg ttgtttctga taagcaacat ccaaatattt tgaccctgct
     8521 tttagtggtt tttttcaaat cttattttga gtcttacttt tagtcataga atagctactg
     8581 atttgatgcg gtctttaact gacttaatat ttttacaatt tcaatatatt ttgcattgga
     8641 atctccagta atgaatatta aaatatatgt acaatcattt gtagatgata tcaattatat
     8701 taagacattt cagatgggct attgtagtat ttaatgtgcc gtattttatg gtagaataat
     8761 tctcagtctc tggacatcaa gattgctttc agtgggaatg aagattaatt tacttcagtc
     8821 ctgatttttt aggcatcaat gcatgttttc atttttgtca gacttttacc ctcttttaat
     8881 gtaattctca acttcttatg gatttacttc ccaatacata aaatccttca aaacaagaat
     8941 gataataatt tttatacttt ttataaaaat aaatttattt ttagtccatc aaggtgtctg
     9001 aagattttat gcctaggtat ctccatatct aacttgataa ggaaaatagg ataaacaatg
     9061 ctggtaatag caggaaagta agtatttgaa taagatgtca aactgatatt tcatgtgaac
     9121 ctaactcatt ttatggtaac taataattat cttatttaaa tcaataggta aaacctgttt
     9181 agaaattaaa aatgagttac gatttaaaga aaattcagat gactcattgt gagtgctagt
     9241 tctcttgtag gatgccactg gaaatgttga aatgaaaaat gtaagtatat cttttggtgg
     9301 aaaaaaggat agtctctagg acacaaaatt actgttttat ttttttctca ggagtttgcc
     9361 taagggtgtg acagatgatc tctgtcactt gtcttagttg tgtcctgcaa taaactggat
     9421 gctttataaa atactagacc tgtgatttcg tatgctgtaa tatttcattt ctccatcacc
     9481 cctccaaatt atttcttagt ttggagtaaa ataataaatg tattatagtc aacatctctt
     9541 gacccctctt tagtttcagc taaactaagc atgtgtgttt gtgtgttcat tttatagttc
     9601 atgtgtagaa ctatgtgaat taaatttaag aaacatgtaa agtagaggaa atagttttct
     9661 ggagaaattt ttcctttttg gatattatgc ccttttccat tgcttttctc tgcttgaaag
     9721 caaaaaaaag taccctaccc ctgttctcct ttagggaaaa actattccta taaagtattt
     9781 ttaaatcgtg caagtcattg cctagggtta gctaaaacat ttctttttaa aaaggagaaa
     9841 atgccctggc tttaacattt tcttgtattt gtatctatta agataaacag tttactttga
     9901 tacagtacat accaatctac ttaatttttt ttccaggatt ccttttacta tgtttggtct
     9961 gaccttttat gataacttaa tattggaaca aattagcata taattctatt ttccatgtga
    10021 cctcaaccag ttgcagaatt gtaccactac tttagggggg gcaatttgac agtttatgta
    10081 gactatagca ttaattgttc ccaaatgttc agtgcatcct ggctaatgtg ttattgaagg
    10141 tgttttcacg taagcagtta gaggaagcac ttcaccccta ttactaagtt attaaaatgc
    10201 ctcctaaagg tagcatttta aattagtata cataattgat tagtaatttg tcttctccca
    10261 agcataaaac agcatagcag agttaagtgt gaccagtgaa gtataagata ttagggattg
    10321 atggtgacaa tgatcatagc aactaaatgg attttttttt tcttttagat tcagccgttg
    10381 gtctttgaaa tttcctgtga tgtgtttcaa tctagatgca aagaacatgg aaaaatcaaa
    10441 gtgctcgagt ggtttaaata tgttttgggt attcctgttt atagactata atacttttcc
    10501 aattaaaatc ctcagttgtc acgcagaaga aggttaagct gtactagtag accacctccc
    10561 ctgcgagcta agctggacag ccaatgacgg gtaagagagt gacatttttc actaacctaa
    10621 gacaggaggg ccgtcagagc tactgcctaa tccaaagacg ggtaaaagtg ataaaaatgt
    10681 atcactccaa cctaagacag gcgcagcttc cgagggattt gagatccaga catgataaga
    10741 tacattgatg agtttggaca aaccaaaact agaatgcagt gaaaaaaatg ccttatttgt
    10801 gaaatttgtg atgctattgc cttatttgta accattataa gctgcaataa acaagtttga
    10861 tatctataac aagaaaatat atatataata agttatcacg taagtagaac atgaaataac
    10921 aatataatta tcgtatgagt taaatcttaa aagtcacgta aaagataatc atgcgtcatt
    10981 ttgactcacg cggtcgttat agttcaaaat cagtgacact taccgcattg acaagcacgc
    11041 ctcacgggag ctccaagcgg cgactgagat gtcctaaatg cacagcgacg gattcgcgct
    11101 atttagaaag agagagcaat atttcaagaa tgcatgcgtc aattttacgc agactatctt
    11161 tctagggtta agaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
    11221 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
    11281 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
    11341 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
    11401 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
    11461 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
    11521 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
    11581 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
    11641 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
    11701 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
    11761 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
    11821 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
    11881 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
    11941 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
    12001 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
    12061 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
    12121 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
    12181 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
    12241 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
    12301 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
    12361 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
    12421 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
    12481 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
    12541 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
    12601 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
    12661 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
    12721 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
    12781 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
    12841 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
    12901 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
    12961 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
    13021 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
    13081 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
    13141 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
    13201 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
    13261 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
    13321 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
    13381 agcctatgga aaaacgccag caacgcggcc tttttacggt ttctggcctt ttgctggcct
    13441 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
    13501 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
    13561 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
    13621 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
    13681 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
    13741 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
    13801 tacgccaagg tcgacttaac cctagaaaga taatcatatt gtgacgtacg ttaaagataa
    13861 tcatgcgtaa aattgacgca tgtgttttat cggtctgtat atcgaggttt atttattaat
    13921 ttgaatagat attaagtttt attatattta cacttacata ctaataataa attcaacaaa
    13981 caatttattt atgtttattt atttattaaa aaaaaacaaa aactcaaaat ttcttctata
    14041 aagtaacaaa acttttagca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
    14101 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
    14161 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
    14221 aaatgtggta tggctgatta tgatcctctg gagatttagc cctcccacac ataaccagag
    14281 ggcagcaatt cacgaatccc aactgccgtc ggctgtccat cactgtcctt cactatggct
    14341 ttgatcccag gatgcagatc gagaagcacc tgtcggcacc gtccgcaggg gctcaagatg
    14401 cccctgttct catttccgat cgcgacgata caagtcaggt tgccagctgc cgcagcagca
    14461 gcagtgccca gcaccacgag ttctgcacaa ggtcccccag taaaatgata tacattgaca
    14521 ccagtgaaga tgcggccgtc gctagagaga gctgcgctgg cgacgctgta gtcttcagag
    14581 atggggatgc tgttgattgt agccgttgct ctttcaatga gggtggattc ttcttgagac
    14641 aaaggcttgg ccatggggcc ggggttctcc tccacgtcgc cggcctgctt cagcaggctg
    14701 aagttggtgg cgccgctgcc cccggggagc atgtcaaggt caaaatcgtc aagagcgtca
    14761 gcaggcagca tatcaaggtc aaagtcgtca agggcatcgg ctgggagcat gtctaagtca
    14821 aaatcgtcaa gggcgtcggt cggcccgccg ctttcgcact ttagctgttt ctccaggcca
    14881 catatgatta gttccaggcc gaaaaggaag gcaggttcgg ctccctgccg gtcgaacagc
    14941 tcaattgctt gtttcagaag tgggggcata gaatcggtgg taggtgtctc tctttcctct
    15001 tttgctactt gatgctcctg ttcctccaat acgcagccca gtgtaaagtg gcccacggcg
    15061 gacagagcgt acagtgcgtt ctccagggag aagccttgct gacacaggaa cgcgagctga
    15121 ttttccaggg tttcgtactg tttctctgtt gggcgggtgc cgagatgcac tttagccccg
    15181 tcgcgatgtg agaggagagc acagcggtat gacttggcgt tgttccgcag aaagtcttgc
    15241 catgactcgc cttccagggg gcaggagtgg gtatgatgcc tgtccagcat ctcgattggc
    15301 agggcatcga gcagggcccg cttgttcttc acgtgccagt acagggtagg ctgctcaact
    15361 cccagctttt gagcgagttt ccttgtcgtc aggccttcga taccgactcc attgagtaat
    15421 tccagagcag agtttatgac tttgctcttg tccagtctag acatcttatc gtcatcgtct
    15481 ttgtaatcca tggtggcgga tcccgcgtca cgacacctgt gttctggcgg caaacccgtt
    15541 gcgaaaaaga acgttcacgg cgactactgc acttatatac ggttctcccc caccctcggg
    15601 aaaaaggcgg agccagtaca cgacatcact ttcccagttt accccgcgcc accttctcta
    15661 ggcaccggtt caattgccga cccctccccc caacttctcg gggactgtgg gcgatgtgcg
    15721 ctctgcccac tgacgggcac cggagccact cgagtggaat t
//
