LOCUS       sequence-348845-       11588 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector plasmid for expressing spCas9-POLI fusion protein and sgRNA. 
            There are two copies of HBB 3' UTR in the 3'UTR of spCas9-POLI to 
            enhance expression. The ST2 loop of the sgRNA scaffold was replace.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11588)
  AUTHORS   Yoo KW, Yadav MK, Song Q, Atala A, Lu B
  TITLE     Targeting DNA polymerase to DNA double-strand breaks reduces DNA 
            deletion size and increases templated insertions generated by 
            CRISPR/Cas9.
  JOURNAL   Nucleic Acids Res. 2022 Mar 22. pii: 6552087. doi: 
            10.1093/nar/gkac186.
  PUBMED    35323942
REFERENCE   2  (bases 1 to 11588)
  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: "Nucleic 
            Acids Res. 2022 Mar 22. pii: 6552087. doi: 10.1093/nar/gkac186."
COMMENT     Sequence Label: pspCas9-POLI-ST2-com vector
FEATURES             Location/Qualifiers
     source          1..11588
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        6..291
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          571..798
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      810..819
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             819..884
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             891..911
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             936..5036
                     /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             936..5036
                     /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             5037..5084
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     RBS             5867..5875
                     /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)"
     primer_bind     complement(8247..8264)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8253..8460
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   8472..8612
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   8472..8601
                     /label=AAV2 ITR
     rep_origin      8687..9142
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8774..8793)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8984..9005
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9159..9178)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9278..9300
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9338..9356)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9424..9528
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9529..9597,9598..10389)
                     /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(9747..9766)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10560..11148
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11049..11068
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        11210..11458
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     11210..11230
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     11381..11400
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 gtacccgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
       61 ccattgacgt caatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      121 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      181 attgacgtca atgacggtaa atggcccgcc tggcattgtg cccagtacat gaccttatgg
      241 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg
      301 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat
      361 ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg
      421 ccrggsgggg sggggggggg sggggggsgg ggsggggsgr ggsggrgrgg kgcggcggca
      481 gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg
      541 ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg
      601 ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
      661 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagctgag caagaggtaa
      721 gggtttaagg gatggttggt tggtggggta ttaatgttta attacctgga gcacctgcct
      781 gaaatcactt tttttcaggt tggaccggtg ccaccatgga ctataaggac cacgacggag
      841 actacaagga tcatgatatt gattacaaag acgatgacga taagatggcc ccaaagaaga
      901 agcggaaggt cggtatccac ggagtcccag cagccgacaa gaagtacagc atcggcctgg
      961 acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca
     1021 agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag
     1081 ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa
     1141 gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga
     1201 tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg
     1261 ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg
     1321 agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg
     1381 acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga
     1441 tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc
     1501 agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg
     1561 ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc
     1621 ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc
     1681 ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca
     1741 cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt
     1801 ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca
     1861 ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc
     1921 aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga
     1981 ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg
     2041 aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc
     2101 tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca
     2161 tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt
     2221 acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct
     2281 actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg
     2341 aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga
     2401 gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca
     2461 agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg
     2521 tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg
     2581 acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca
     2641 agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct
     2701 ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg
     2761 aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag
     2821 agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc
     2881 agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca
     2941 tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca
     3001 acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga
     3061 aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca
     3121 gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag
     3181 tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca
     3241 cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag
     3301 agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga
     3361 agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca
     3421 tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg
     3481 actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg
     3541 tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca
     3601 agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg
     3661 aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc
     3721 acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga
     3781 tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt
     3841 tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga
     3901 acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt
     3961 acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg
     4021 gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga
     4081 ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg
     4141 gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc
     4201 cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta
     4261 tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga
     4321 agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg
     4381 aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg
     4441 aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag
     4501 tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc
     4561 ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct
     4621 ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg
     4681 aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca
     4741 tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag
     4801 tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca
     4861 tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca
     4921 ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc
     4981 agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacaaaa
     5041 ggccggcggc cacgaaaaag gccggccagg caaaaaagaa aaagggatcc tctgagacac
     5101 caggcacctc cgagtctgcc acccctgagg gaggaagcgg aggaagcggc tccgtgcaga
     5161 tcccacagaa ccccctgatc ctggtggacg gcagctccta cctgtatcgg gcctaccacg
     5221 ccttcccacc tctgacaaac tccgccggag agccaaccgg agccatgtat ggcgtgctga
     5281 atatgctgag gagcctgatc atgcagtaca agcctacaca cgccgccgtg gtgtttgatg
     5341 ccaagggcaa gaccttccgc gacgagctgt ttgagcacta caagagccac aggccaccaa
     5401 tgcctgacga tctgagggca cagatcgagc cactgcacgc aatggtgaag gccatgggcc
     5461 tgcctctgct ggccgtgagc ggagtggagg ccgacgatgt gatcggcaca ctggcaaggg
     5521 aggcagagaa ggcaggccgc ccagtgctga tctccaccgg cgacaaggat atggcccagc
     5581 tggtgacacc aaacatcacc ctgatcaaca ccatgacaaa taccatcctg ggccccgagg
     5641 aggtggtgaa taagtatggc gtgcctccag agctgatcat cgatttcctg gccctgatgg
     5701 gcgactctag cgataacatc cctggcgtgc caggagtggg agaaaagacc gcacaggccc
     5761 tgctgcaggg cctgggaggc ctggacaccc tgtacgccga gccagagaag atcgccggcc
     5821 tgtcctttag gggcgccaag acaatggccg ccaagctgga gcagaataag gaggtggcct
     5881 acctgtctta tcagctggcc acaatcaaga ccgacgtgga gctggagctg acctgcgagc
     5941 agctggaggt gcagcagcct gcagcagagg agctgctggg cctgttcaag aagtacgagt
     6001 ttaagagatg gacagccgat gtggaggccg gcaagtggct gcaggcaaag ggagcaaagc
     6061 cagcagcaaa gccacaggag acaagcgtgg cagacgaggc accagaggtg acagccaccg
     6121 tgatctccta cgataactat gtgacaatcc tggacgagga gacactgaag gcctggatcg
     6181 ccaagctgga gaaggccccc gtgttcgcct ttgatacaga gacagacagc ctggataaca
     6241 tctccgccaa tctggtgggc ctgtctttcg ccatcgagcc tggcgtggcc gcctatatcc
     6301 cagtggccca cgactacctg gatgcccccg accagatcag cagggagaga gccctggagc
     6361 tgctgaagcc tctgctggag gatgagaagg ccctgaaggt cggccagaac ctgaagtatg
     6421 acaggggcat cctggccaat tacggcatcg agctgagagg catcgccttt gacaccatgc
     6481 tggagtctta tatcctgaat agcgtggcag gccggcacga catggattcc ctggccgaga
     6541 ggtggctgaa gcacaagaca atcaccttcg aggagatcgc cggcaagggc aagaaccagc
     6601 tgaccttcaa ccagatcgcc ctggaggagg caggcaggta cgcagcagag gacgcagatg
     6661 tgaccctgca gctgcacctg aagatgtggc cagatctgca gaagcacaag ggccccctga
     6721 acgtctttga gaatatcgag atgcccctgg tgcctgtgct gagccggatc gagcgcaacg
     6781 gcgtgaagat cgaccctaag gtgctgcaca atcactccga ggagctgacc ctgagactgg
     6841 ccgagctgga gaagaaggcc cacgagatcg ccggcgagga gttcaacctg tcctctacaa
     6901 agcagctgca gaccatcctg tttgagaagc agggcatcaa gcccctgaag aaaacccctg
     6961 gaggagcacc atctaccagc gaggaggtgc tggaggagct ggccctggat tatcccctgc
     7021 ctaaagtgat cctggagtac cggggcctgg ccaagctgaa gtctacatat accgacaagc
     7081 tgcccctgat gatcaaccct aagacaggcc gggtgcacac cagctaccac caggcagtga
     7141 cagcaaccgg ccgcctgagc tccaccgatc caaacctgca gaatatcccc gtgaggaatg
     7201 aggagggcag gagaatcaga caggccttca tcgcccccga ggattatgtg atcgtgtccg
     7261 ccgactactc tcagatcgag ctgaggatca tggcccacct gtccagagat aagggcctgc
     7321 tgacagcctt cgccgagggc aaggacatcc acagggcaac cgcagcagag gtgtttggcc
     7381 tgcctctgga gacagtgacc tccgagcagc ggcgctctgc caaggccatc aacttcggcc
     7441 tgatctatgg catgtctgcc tttggcctgg ccaggcagct gaatatccct agaaaggagg
     7501 cccagaagta catggacctg tatttcgaga gataccccgg cgtgctggag tacatggaga
     7561 ggacacgcgc acaggcaaag gagcagggct atgtggagac actggatggc aggagactgt
     7621 acctgccaga catcaagtct agcaacggag caaggagggc agcagcagag agggccgcca
     7681 tcaatgcccc catgcagggc acagccgccg atatcatcaa gagagccatg atcgcagtgg
     7741 acgcctggct gcaggcagag cagccaaggg tgagaatgat catgcaggtg cacgatgagc
     7801 tggtgtttga ggtgcacaag gacgatgtgg acgccgtggc caagcagatc caccagctga
     7861 tggagaactg tacccgcctg gatgtgccac tgctggtgga agtgggaagc ggagagaatt
     7921 gggaccaggc ccactgagga tcctaagctc gctttcttgc tgtccaattt ctattaaagg
     7981 ttcctttgtt ccctaagtcc aactactaaa ctgggggata ttatgaaggg ccttgagcat
     8041 ctggattctg cctaataaaa aacatttatt ttcattgcta gctcgctttc ttgctgtcca
     8101 atttctatta aaggttcctt tgttccctaa gtccaactac taaactgggg gatattatga
     8161 agggccttga gcatctggat tctgcctaat aaaaaacatt tattttcatt gcgaattcta
     8221 gaaaattcct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     8281 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     8341 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     8401 tggggtgggg caggacagca agggggagga ttgggaagag aatagcaggc atgctgggga
     8461 ggtacggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     8521 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     8581 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     8641 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     8701 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     8761 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     8821 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     8881 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     8941 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     9001 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     9061 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     9121 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     9181 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     9241 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     9301 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     9361 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     9421 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     9481 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     9541 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     9601 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     9661 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     9721 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     9781 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     9841 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     9901 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     9961 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
    10021 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
    10081 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
    10141 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
    10201 gctggctggt ttattgctga taaatctgga gccggtgagc gtggaagccg cggtatcatt
    10261 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
    10321 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
    10381 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
    10441 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
    10501 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
    10561 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
    10621 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
    10681 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
    10741 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
    10801 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
    10861 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
    10921 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
    10981 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
    11041 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
    11101 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
    11161 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtg agggcctatt
    11221 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat
    11281 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
    11341 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
    11401 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg
    11461 tgtcttcctc gaggaagacc cgtttgagag ctagaaatag caagttcaaa taaggctagt
    11521 ccgttatcaa cttggctgaa tgcctgcgag catcccaccc aagtggcacc gagtcggtgc
    11581 tttttttg
//
