LOCUS       sequence-343617-       10215 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SpCas9 prime editor 2 with P2A-BSD marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10215)
  AUTHORS   Chen PJ, Hussmann JA, Yan J, Knipping F, Ravisankar P, Chen PF, Chen
            C, Nelson JW, Newby GA, Sahin M, Osborn MJ, Weissman JS, Adamson B, 
            Liu DR
  TITLE     Enhanced prime editing systems by manipulating cellular determinants
            of editing outcomes.
  JOURNAL   Cell. 2021 Oct 28;184(22):5635-5652.e29. doi: 
            10.1016/j.cell.2021.09.018.
  PUBMED    34653350
REFERENCE   2  (bases 1 to 10215)
  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.cell.2021.09"; journalName: "Cell"; date: "2021-10-28- 
            28"; volume: "184"; issue: "22"; pages: "5635-5652.e29"
COMMENT     Sequence Label: pCMV-PE2-P2A-BSD
FEATURES             Location/Qualifiers
     source          1..10215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        42..421
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        422..625
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     576..596
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     667..686
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        667..685
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             735..755
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(756..3269,3270..3272,3273..4856)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the H840A mutation in the HNH catalytic domain"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             756..4856
                     /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
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7029..7049
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7059..7115
                     /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             7116..7514
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             7520..7537
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7560..7577)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7566..7790
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7861..7877)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7861..7877)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7874..7896)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7885..7901
                     /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(7909..7915,7916..7933,7934..7939))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7954..7975
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8092..8109)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8263..8851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8343..8362)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9022..9813,9814..9882))
                     /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     9645..9664
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9883..9987)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10066..10085)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
       61 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      121 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      181 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      241 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      301 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      361 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      421 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      481 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      541 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      601 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agagatccgc
      661 ggccgctaat acgactcact atagggagag ccgccaccat gaaacggaca gccgacggaa
      721 gcgagttcga gtcaccaaag aagaagcgga aagtcgacaa gaagtacagc atcggcctgg
      781 acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca
      841 agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag
      901 ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa
      961 gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga
     1021 tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg
     1081 ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg
     1141 agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg
     1201 acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga
     1261 tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc
     1321 agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg
     1381 ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc
     1441 ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc
     1501 ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca
     1561 cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt
     1621 ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca
     1681 ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc
     1741 aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga
     1801 ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg
     1861 aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc
     1921 tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca
     1981 tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt
     2041 acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct
     2101 actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg
     2161 aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga
     2221 gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca
     2281 agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg
     2341 tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg
     2401 acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca
     2461 agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct
     2521 ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg
     2581 aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag
     2641 agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc
     2701 agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca
     2761 tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca
     2821 acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga
     2881 aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca
     2941 gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag
     3001 tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca
     3061 cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag
     3121 agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga
     3181 agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca
     3241 tcaaccggct gtccgactac gatgtggacg ctatcgtgcc tcagagcttt ctgaaggacg
     3301 actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg
     3361 tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca
     3421 agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg
     3481 aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc
     3541 acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga
     3601 tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt
     3661 tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga
     3721 acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt
     3781 acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg
     3841 gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga
     3901 ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg
     3961 gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc
     4021 cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta
     4081 tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga
     4141 agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg
     4201 aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg
     4261 aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag
     4321 tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc
     4381 ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct
     4441 ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg
     4501 aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca
     4561 tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag
     4621 tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca
     4681 tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca
     4741 ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc
     4801 agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga ggtgactctg
     4861 gaggatctag cggaggatcc tctggcagcg agacaccagg aacaagcgag tcagcaacac
     4921 cagagagcag tggcggcagc agcggcggca gcagcaccct aaatatagaa gatgagtatc
     4981 ggctacatga gacctcaaaa gagccagatg tttctctagg gtccacatgg ctgtctgatt
     5041 ttcctcaggc ctgggcggaa accgggggca tgggactggc agttcgccaa gctcctctga
     5101 tcatacctct gaaagcaacc tctacccccg tgtccataaa acaatacccc atgtcacaag
     5161 aagccagact ggggatcaag ccccacatac agagactgtt ggaccaggga atactggtac
     5221 cctgccagtc cccctggaac acgcccctgc tacccgttaa gaaaccaggg actaatgatt
     5281 ataggcctgt ccaggatctg agagaagtca acaagcgggt ggaagacatc caccccaccg
     5341 tgcccaaccc ttacaacctc ttgagcgggc tcccaccgtc ccaccagtgg tacactgtgc
     5401 ttgatttaaa ggatgccttt ttctgcctga gactccaccc caccagtcag cctctcttcg
     5461 cctttgagtg gagagatcca gagatgggaa tctcaggaca attgacctgg accagactcc
     5521 cacagggttt caaaaacagt cccaccctgt ttaatgaggc actgcacaga gacctagcag
     5581 acttccggat ccagcaccca gacttgatcc tgctacagta cgtggatgac ttactgctgg
     5641 ccgccacttc tgagctagac tgccaacaag gtactcgggc cctgttacaa accctaggga
     5701 acctcgggta tcgggcctcg gccaagaaag cccaaatttg ccagaaacag gtcaagtatc
     5761 tggggtatct tctaaaagag ggtcagagat ggctgactga ggccagaaaa gagactgtga
     5821 tggggcagcc tactccgaag acccctcgac aactaaggga gttcctaggg aaggcaggct
     5881 tctgtcgcct cttcatccct gggtttgcag aaatggcagc ccccctgtac cctctcacca
     5941 aaccggggac tctgtttaat tggggcccag accaacaaaa ggcctatcaa gaaatcaagc
     6001 aagctcttct aactgcccca gccctggggt tgccagattt gactaagccc tttgaactct
     6061 ttgtcgacga gaagcagggc tacgccaaag gtgtcctaac gcaaaaactg ggaccttggc
     6121 gtcggccggt ggcctacctg tccaaaaagc tagacccagt agcagctggg tggccccctt
     6181 gcctacggat ggtagcagcc attgccgtac tgacaaagga tgcaggcaag ctaaccatgg
     6241 gacagccact agtcattctg gccccccatg cagtagaggc actagtcaaa caaccccccg
     6301 accgctggct ttccaacgcc cggatgactc actatcaggc cttgcttttg gacacggacc
     6361 gggtccagtt cggaccggtg gtagccctga acccggctac gctgctccca ctgcctgagg
     6421 aagggctgca acacaactgc cttgatatcc tggccgaagc ccacggaacc cgacccgacc
     6481 taacggacca gccgctccca gacgccgacc acacctggta cacggatgga agcagtctct
     6541 tacaagaggg acagcgtaag gcgggagctg cggtgaccac cgagaccgag gtaatctggg
     6601 ctaaagccct gccagccggg acatccgctc agcgggctga actgatagca ctcacccagg
     6661 ccctaaagat ggcagaaggt aagaagctaa atgtttatac tgatagccgt tatgcttttg
     6721 ctactgccca tatccatgga gaaatataca gaaggcgtgg gtggctcaca tcagaaggca
     6781 aagagatcaa aaataaagac gagatcttgg ccctactaaa agccctcttt ctgcccaaaa
     6841 gacttagcat aatccattgt ccaggacatc aaaagggaca cagcgccgag gctagaggca
     6901 accggatggc tgaccaagcg gcccgaaagg cagccatcac agagactcca gacacctcta
     6961 ccctcctcat agaaaattca tcaccctctg gcggctcaaa aagaaccgcc gacggcagcg
     7021 aattcgagcc caagaagaag aggaaagtcg gatccggcgc aacaaacttc tctctgctga
     7081 aacaagccgg agatgtcgaa gagaatcctg gaccgatggc caagcctttg tctcaagaag
     7141 aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact
     7201 acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat
     7261 atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg
     7321 cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc
     7381 cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga
     7441 aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg
     7501 tgtgggaggg ctaaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     7561 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     7621 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgagaaaa ttgcatcgca
     7681 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     7741 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     7801 ggaaagaacc agctggggct cgataccgtc gacctctagc tagagcttgg cgtaatcatg
     7861 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     7921 cggaagcata aagtgtaaag cctagggtgc ctaatgagtg agctaactca cattaattgc
     7981 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     8041 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     8101 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     8161 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     8221 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     8281 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     8341 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     8401 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     8461 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     8521 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     8581 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     8641 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     8701 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     8761 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     8821 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     8881 tgacactcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     8941 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     9001 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     9061 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     9121 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     9181 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     9241 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     9301 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     9361 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     9421 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     9481 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     9541 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     9601 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     9661 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     9721 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     9781 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     9841 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     9901 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     9961 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
    10021 cggatcggga gatcgatctc ccgatcccct agggtcgact ctcagtacaa tctgctctga
    10081 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
    10141 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
    10201 gcttagggtt aggcg
//
