LOCUS       sequence-388882-       13491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized SpCas9-ABE8e(V106W) and blasticidin 
            resistance: EFS promoter-SpCas9-ABE8e(V106W)-NLS-FLAG-P2A-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13491)
  AUTHORS   Kim N, Choi S, Kim S, Song M, Seo JH, Min S, Park J, Cho SR, Kim HH
  TITLE     Deep learning models to predict the editing efficiencies and 
            outcomes of diverse base editors.
  JOURNAL   Nat Biotechnol. 2024 Mar;42(3):484-497. doi: 
            10.1038/s41587-023-01792-x
  PUBMED    37188916
REFERENCE   2  (bases 1 to 13491)
  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"; journalName: 
            "Nat Biotechnol"; date: "2024-03"; volume: "42"; issue: "3"; pages: 
            "484-497"
COMMENT     Sequence Label: SpCas9-ABE8e(V106W)
FEATURES             Location/Qualifiers
     source          1..13491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    417..538
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(454..473)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     508..527
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(587..603)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(587..603)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(600..622)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    611..627
                     /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(635..641,642..659,660..665))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    680..701
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(818..835)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(989..1577)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1069..1088)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1748..2539,2540..2608))
                     /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     2371..2390
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2609..2713)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2788..2807)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2979..3358
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3360..3558
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3513..3533
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3576..3756
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3803..3928
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4421..4654
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4839..4883
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5181..5298
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5391..5602
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             5653..5673
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(6268..6291,6292..6294,6295..10368)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6268..10368
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             10411..10431
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10432..10455
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             10465..10521
                     /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             10522..10920
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    10945..11533
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10998..11018)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11416..11427)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(11536..11552)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11537..11553)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             12058..12238
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(12264..12281)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12270..12494
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      12540..12968
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12627..12646)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12837..12858
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12977..12997)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      13142..13276
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     13203..13222
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        13338..13385
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(13404..13491,1..287)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
ORIGIN
        1 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
       61 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
      121 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
      181 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
      241 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
      301 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
      361 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
      421 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
      481 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
      541 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
      601 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
      661 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
      721 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
      781 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
      841 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
      901 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
      961 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1021 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1081 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1141 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1201 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1261 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1321 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1381 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1441 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1501 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1561 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1621 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1681 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1741 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     1801 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     1861 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     1921 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     1981 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     2041 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     2101 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     2161 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     2221 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     2281 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     2341 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     2401 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     2461 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     2521 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     2581 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     2641 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2701 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     2761 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     2821 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     2881 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     2941 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
     3001 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     3061 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     3121 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     3181 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     3241 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     3301 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     3361 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     3421 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     3481 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     3541 ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct
     3601 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     3661 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     3721 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     3781 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     3841 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     3901 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     3961 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     4021 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     4081 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     4141 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     4201 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     4261 gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     4321 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     4381 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     4441 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     4501 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     4561 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     4621 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     4681 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     4741 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     4801 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     4861 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     4921 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
     4981 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
     5041 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
     5101 gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa
     5161 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     5221 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     5281 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttggttaatt
     5341 agctagctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg
     5401 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgat ccggtgccta
     5461 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
     5521 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
     5581 cgggtttgcc gccagaacac aggaccggtt gccaccatga aacggacagc cgacggaagc
     5641 gagttcgagt caccaaagaa gaagcggaaa gtctctgagg tggagttttc ccacgagtac
     5701 tggatgagac atgccctgac cctggccaag agggcacggg atgagaggga ggtgcctgtg
     5761 ggagccgtgc tggtgctgaa caatagagtg atcggcgagg gctggaacag agccatcggc
     5821 ctgcacgacc caacagccca tgccgaaatt atggccctga gacagggcgg cctggtcatg
     5881 cagaactaca gactgattga cgccaccctg tacgtgacat tcgagccttg cgtgatgtgc
     5941 gccggcgcca tgatccactc taggatcggc cgcgtggtgt ttggatggag aaattctaaa
     6001 agaggcgccg caggctccct gatgaacgtg ctgaactacc ccggcatgaa tcaccgcgtc
     6061 gaaattaccg agggaatcct ggcagatgaa tgtgccgccc tgctgtgcga tttctatcgg
     6121 atgcctagac aggtgttcaa tgctcagaag aaggcccaga gctccatcaa ctccggagga
     6181 tctagcggag gctcctctgg ctctgagaca cctggcacaa gcgagagcgc aacacctgaa
     6241 agcagcgggg gcagcagcgg ggggtcagac aagaagtaca gcatcggcct ggccatcggc
     6301 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
     6361 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg
     6421 ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac
     6481 accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag
     6541 gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag
     6601 cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
     6661 cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg
     6721 ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc
     6781 gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac
     6841 aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg
     6901 tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag
     6961 aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc
     7021 aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac
     7081 gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc
     7141 gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc
     7201 accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg
     7261 accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc
     7321 gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc
     7381 tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag
     7441 ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac
     7501 cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc
     7561 ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg
     7621 ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc
     7681 atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc
     7741 gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc
     7801 ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag
     7861 ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg
     7921 ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc
     7981 gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc
     8041 acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac
     8101 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
     8161 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
     8221 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
     8281 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
     8341 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
     8401 gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc
     8461 attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc
     8521 cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag
     8581 ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc
     8641 agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac
     8701 ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg
     8761 ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc
     8821 gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc
     8881 gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt
     8941 acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat
     9001 aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca
     9061 cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa
     9121 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt
     9181 tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc
     9241 gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac
     9301 tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct
     9361 accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg
     9421 gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc
     9481 gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg
     9541 aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc
     9601 aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc
     9661 ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc
     9721 aagtccaaga aactgaagag tgtgaaagag ctactgggga tcaccatcat ggaaagaagc
     9781 agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag
     9841 gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga
     9901 atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat
     9961 gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat
    10021 gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag
    10081 atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc
    10141 gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg
    10201 tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac
    10261 cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc
    10321 accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgactc tggcggctca
    10381 aaaagaaccg ccgacggcag cgaattcgag cccaagaaga agaggaaagt cgattacaaa
    10441 gacgatgacg ataagggatc cggcgcaaca aacttctctc tgctgaaaca agccggagat
    10501 gtcgaagaga atcctggacc gatggccaag cctttgtctc aagaagaatc caccctcatt
    10561 gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag cgtcgccagc
    10621 gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca ttttactggg
    10681 ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc tggcaacctg
    10741 acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg cggacggtgc
    10801 cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga cagtgatgga
    10861 cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa
    10921 gaattcgata tcaagcttat cggtaatcaa cctctggatt acaaaatttg tgaaagattg
    10981 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
    11041 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
    11101 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
    11161 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
    11221 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
    11281 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
    11341 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
    11401 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
    11461 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
    11521 gccgcctccc cgcatcgata ccgtcgacct cgagacctag aaaaacatgg agcaatcaca
    11581 agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag
    11641 gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct
    11701 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
    11761 cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt ccctgattgg
    11821 cagaactaca caccagggcc agggatcaga tatccactga cctttggatg gtgctacaag
    11881 ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg aaggagagaa cacccgcttg
    11941 ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtatt agagtggagg
    12001 tttgacagcc gcctagcatt tcatcacatg gcccgagagc tgcatccgga ctgtactggg
    12061 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
    12121 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
    12181 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagg
    12241 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
    12301 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
    12361 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
    12421 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
    12481 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
    12541 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
    12601 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
    12661 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
    12721 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
    12781 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
    12841 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
    12901 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
    12961 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
    13021 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
    13081 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
    13141 gtcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
    13201 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
    13261 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
    13321 ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat
    13381 acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg
    13441 cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc t
//
