LOCUS       sequence-329029-        9232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcription of TREX1 guide RNA and Cas9 expression for CRISPR/Cas9
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9232)
  AUTHORS   Mohr L, Toufektchan E, von Morgen P, Chu K, Kapoor A, Maciejowski J
  TITLE     ER-directed TREX1 limits cGAS activation at micronuclei.
  JOURNAL   Mol Cell. 2021 Jan 12. pii: S1097-2765(20)30958-8. doi: 
            10.1016/j.molcel.2020.12.037.
  PUBMED    33476576
REFERENCE   2  (bases 1 to 9232)
  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: "Mol Cell. 
            2021 Jan 12. pii: S1097-2765(20)30958-8. doi: 
            10.1016/j.molcel.2020.12.037."
COMMENT     Sequence Label: pU6-TREX1 g3 Cas9-T2A-mCherry
FEATURES             Location/Qualifiers
     source          1..9232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          133..360
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      372..381
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             381..446
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             453..473
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             498..4598
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             498..4598
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4599..4646
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4650..4703
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4710..5420
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4855..4873)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(5445..5462)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5451..5658
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5667..5807
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5667..5796
                     /label=AAV2 ITR
     rep_origin      5882..6337
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5969..5988)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6179..6200
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6354..6373)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6473..6495
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6533..6551)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6619..6723
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6724..6792,6793..7584)
                     /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(6942..6961)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7755..8343
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8244..8263
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8405..8653
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8405..8425
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8576..8595
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8671..8746
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8843..9128
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ggcggggggn nnnnnngggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg
       61 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
      121 gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc
      181 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
      241 ggcccttctc ctccgggctg taattagctg agcaagaggt aagggtttaa gggatggttg
      301 gttggtgggg tattaatgtt taattacctg gagcacctgc ctgaaatcac tttttttcag
      361 gttggaccgg tgccaccatg gactataagg accacgacgg agactacaag gatcatgata
      421 ttgattacaa agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcggtatcc
      481 acggagtccc agcagccgac aagaagtaca gcatcggcct ggacatcggc accaactctg
      541 tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg
      601 gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg
      661 gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga
      721 agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca
      781 gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc
      841 accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct
      901 accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc
      961 tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc
     1021 ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt
     1081 tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac
     1141 tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg
     1201 gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact
     1261 tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg
     1321 acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc
     1381 tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc
     1441 ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga
     1501 aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca
     1561 agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca
     1621 tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag
     1681 aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc
     1741 tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca
     1801 accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg
     1861 ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct
     1921 ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga
     1981 ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg
     2041 agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa
     2101 agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca
     2161 accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg
     2221 actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg
     2281 atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc
     2341 tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc
     2401 tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat
     2461 acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg
     2521 gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc
     2581 tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc
     2641 agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg
     2701 gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc
     2761 ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga
     2821 acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc
     2881 tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc
     2941 tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact
     3001 acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg
     3061 tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg
     3121 tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa
     3181 agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct
     3241 tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg
     3301 actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga
     3361 tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc
     3421 gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg
     3481 ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt
     3541 acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt
     3601 acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg
     3661 agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata
     3721 agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga
     3781 aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca
     3841 gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca
     3901 gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga
     3961 aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga
     4021 agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca
     4081 tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct
     4141 ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc
     4201 tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac
     4261 agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt
     4321 tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca
     4381 agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga
     4441 ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt
     4501 acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt
     4561 acgagacacg gatcgacctg tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa
     4621 aggccggcca ggcaaaaaag aaaaagcttg agggcagagg aagtctgcta acatgcggtg
     4681 acgtggagga gaatcccggc cctgctagca tggtgagcaa gggcgaggag gataacatgg
     4741 ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg
     4801 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     4861 tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca
     4921 tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt
     4981 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga
     5041 ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg
     5101 gcaccaactt cccctcagac ggccccgtaa tgcagaagaa aaccatgggc tgggaggcct
     5161 cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga
     5221 agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc
     5281 ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg
     5341 aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca
     5401 tggacgagct gtacaagtaa gaattcctag agctcgctga tcagcctcga ctgtgccttc
     5461 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     5521 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     5581 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagagaa
     5641 tagcaggcat gctggggagc ggccgcagga acccctagtg atggagttgg ccactccctc
     5701 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     5761 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     5821 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     5881 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     5941 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     6001 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     6061 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     6121 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     6181 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     6241 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
     6301 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     6361 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     6421 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     6481 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     6541 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     6601 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     6661 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     6721 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     6781 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     6841 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     6901 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6961 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     7021 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     7081 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     7141 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     7201 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     7261 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     7321 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     7381 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtgga
     7441 agccgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     7501 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     7561 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     7621 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     7681 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     7741 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     7801 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     7861 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     7921 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7981 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     8041 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     8101 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     8161 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     8221 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     8281 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     8341 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     8401 atgtgagggc ctatttccca tgattccttc atatttgcat atacgataca aggctgttag
     8461 agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg
     8521 tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta
     8581 tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa
     8641 aggacgaaac accgagcccc cccacctctc gttttagagc tagaaatagc aagttaaaat
     8701 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
     8761 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
     8821 gacaaatggc tctagaggta cccgttacat aacttacggt aaatggcccg cctggctgac
     8881 cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact ttccattgac
     8941 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     9001 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattgtgccc
     9061 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     9121 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
     9181 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg gg
//
