LOCUS       sequence-388917-       14391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized SpCas9-NRCH-CGBE1 and blasticidin 
            resistance: EFS promoter-SpCas9-NRCH-CGBE1-NLS-FLAG-P2A-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14391)
  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 14391)
  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-NRCH-CGBE1
FEATURES             Location/Qualifiers
     source          1..14391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..498
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        500..698
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     653..673
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             716..896
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    943..1068
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1561..1794
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1979..2023
                     /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    2321..2438
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2531..2742
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2793..2813
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4308..8408
                     /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
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMVKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGIIPHQIHLGELHAILRRQGDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRLRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGGH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKGNSDKLIARKKDWDPKKYGGFNSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTINRKQYNTTKEVLD
                     ATLIRQSITGLYETRIDLSQLGGD"
     CDS             8451..8471
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8472..8495
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8505..8561
                     /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             8562..8960
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8985..9573
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9038..9058)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9456..9467)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9576..9592)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9577..9593)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10098..10278
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10304..10321)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10310..10534
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10580..11008
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10667..10686)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10877..10898
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11017..11037)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      11182..11316
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11243..11262
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11378..11425
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11444..11818
                     /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"
     polyA_signal    11948..12069
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11985..12004)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12039..12058
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12118..12134)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12118..12134)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12131..12153)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12142..12158
                     /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(12166..12172,12173..12190,12191..12196))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12211..12232
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12349..12366)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12520..13108)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12600..12619)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13279..14070,14071..14139))
                     /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     13902..13921
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14140..14244)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14319..14338)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
       61 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
      121 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      181 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      241 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      301 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      361 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      421 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      481 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      541 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      601 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      661 ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg cgttttgcct gtactgggtc
      721 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
      781 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
      841 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
      901 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
      961 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
     1021 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
     1081 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
     1141 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
     1201 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
     1261 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
     1321 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
     1381 aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat
     1441 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     1501 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     1561 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     1621 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     1681 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     1741 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     1801 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     1861 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     1921 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     1981 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     2041 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     2101 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     2161 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
     2221 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg
     2281 tgcgccaatt ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
     2341 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     2401 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
     2461 gagatccagt ttggttaatt agctagctag gtcttgaaag gagtgggaat tggctccggt
     2521 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     2581 ggcaattgat ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     2641 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     2701 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggaccggtt gccaccatga
     2761 aacggacagc cgacggaagc gagttcgagt caccaaagaa gaagcggaaa gtcgccaacg
     2821 aactgacctg gcacgacgtg ctggccgaag agaagcagca gccctatttc cttaacaccc
     2881 tgcagaccgt ggccagcgag cggcagagcg gcgtgaccat ctacccccca cagaaagacg
     2941 tgtttaacgc cttccgcttc acagagctgg gcgacgtgaa ggtggtgatc ctgggccagg
     3001 acccctacca cggccccggc caggcccatg gtctggcctt cagcgtgcgg cccggcatcg
     3061 ccatcccccc cagcctgctg aacatgtata aggagctgga aaacaccatc cccggcttca
     3121 cccggcccaa tcacggctac ctggagagct gggcgcggca gggcgtgctg ctgctcaaca
     3181 ccgtgctgac ggtacgggcc ggccaggcgc atagccacgc cagcctgggc tgggagacct
     3241 tcaccgataa ggtgatcagc ctgatcaacc agcaccggga gggcgtggtg tttctgctgt
     3301 ggggcagcca tgcccagaag aaaggcgcca tcatcgacaa gcagcggcat catgtgctga
     3361 aagcccccca ccccagcccc cttagcgccc accggggctt cttcggctgc aaccatttcg
     3421 tgctggccaa tcagtggctg gaacaacggg gcgagacccc cattgactgg atgcccgtgt
     3481 tgcccgccga gagcgagagc ggcgggagcg gcgggagcgg ggggagctcc tcagagactg
     3541 ggcctgtcgc cgtcgatcca accctgcgcc gccggattga acctcacgag tttgaagtgt
     3601 tctttgaccc ccgggagctg gccaaggaga catgcctgct gtacgagatc aactggggag
     3661 gcaggcactc catctggagg cacacctctc agaacacaaa taagcacgtg gaggtgaact
     3721 tcatcgagaa gtttaccaca gagcggtact tctgccccaa taccagatgt agcatcacat
     3781 ggtttctgag ctggtcccct tgcggagagt gtagcagggc catcaccgag ttcctgtcca
     3841 gatatccaca cgtgacactg tttatctaca tcgccaggct gtatcaccac gcagacccaa
     3901 ggaataggca gggcctgcgc gatctgatca gctccggcgt gaccatccag atcatgacag
     3961 agcaggagtc cggctactgc tggcggaact tcgtgaatta ttctcctagc aacgaggccc
     4021 actggcctag gtacccacac ctgtgggtgc gcctgtacgt gctggagctg tattgcatca
     4081 tcctgggcct gcccccttgt ctgaatatcc tgcggagaaa gcagccccag ctgaccttct
     4141 ttacaatcgc cctgcagtct tgtcactatc agaggctgcc accccacatc ctgtgggcca
     4201 caggcctgaa gtctggagga tctagcggag gatcctctgg cagcgagaca ccaggaacaa
     4261 gcgagtcagc aacaccagag agcagtggcg gcagcagcgg cggcagcgac aagaagtaca
     4321 gcatcggcct ggccatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca
     4381 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga
     4441 acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga
     4501 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct
     4561 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc
     4621 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg
     4681 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca
     4741 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg
     4801 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca
     4861 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg
     4921 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga
     4981 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc
     5041 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc
     5101 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt
     5161 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc
     5221 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatggtg aagagatacg
     5281 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga
     5341 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg
     5401 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca
     5461 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg
     5521 acaacggcat catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc
     5581 agggcgattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct
     5641 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga
     5701 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg
     5761 cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga
     5821 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca
     5881 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa
     5941 aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag
     6001 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc
     6061 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact
     6121 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt
     6181 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca
     6241 aagtgatgaa gcagctgaag cggctgagat acaccggctg gggcaggctg agccggaagc
     6301 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg
     6361 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag
     6421 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca
     6481 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg
     6541 agctcgtgaa agtgatgggc ggccacaagc ccgagaacat cgtgatcgaa atggccagag
     6601 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag
     6661 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc
     6721 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc
     6781 aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct
     6841 ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca
     6901 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc
     6961 tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag
     7021 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc
     7081 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga
     7141 atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
     7201 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg
     7261 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa
     7321 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg
     7381 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt
     7441 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa
     7501 acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag
     7561 tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca
     7621 gcaaagagtc tatcctgccc aagggcaaca gcgataagct gatcgccaga aagaaggact
     7681 gggaccctaa gaagtacggc ggcttcaaca gccccaccgt ggcctattct gtgctggtgg
     7741 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctactgggga
     7801 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg
     7861 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc
     7921 tggaaaacgg ccggaagaga atgctggcct ctgccggcgt gctgcagaag ggaaacgaac
     7981 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga
     8041 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc
     8101 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta
     8161 atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg
     8221 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt
     8281 actttgacac caccatcaac cggaagcagt acaacaccac caaagaggtg ctggacgcca
     8341 ccctgatccg gcagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg
     8401 gaggcgactc tggcggctca aaaagaaccg ccgacggcag cgaattcgag cccaagaaga
     8461 agaggaaagt cgattacaaa gacgatgacg ataagggatc cggcgcaaca aacttctctc
     8521 tgctgaaaca agccggagat gtcgaagaga atcctggacc gatggccaag cctttgtctc
     8581 aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg
     8641 aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca
     8701 atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg
     8761 ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct
     8821 tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca
     8881 tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg
     8941 gttatgtgtg ggagggctaa gaattcgata tcaagcttat cggtaatcaa cctctggatt
     9001 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     9061 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     9121 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     9181 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     9241 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     9301 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     9361 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     9421 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     9481 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     9541 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct cgagacctag
     9601 aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc
     9661 tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac
     9721 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     9781 aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac
     9841 aaggctactt ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga
     9901 cctttggatg gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg
     9961 aaggagagaa cacccgcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
    10021 gagaagtatt agagtggagg tttgacagcc gcctagcatt tcatcacatg gcccgagagc
    10081 tgcatccgga ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
    10141 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
    10201 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
    10261 tgtggaaaat ctctagcagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
    10321 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
    10381 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
    10441 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
    10501 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
    10561 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
    10621 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
    10681 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
    10741 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
    10801 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
    10861 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
    10921 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
    10981 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
    11041 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
    11101 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
    11161 tcaattagtc agcaaccata gtcccgcccc taactccgcc cagttccgcc cattctccgc
    11221 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
    11281 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
    11341 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
    11401 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
    11461 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
    11521 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
    11581 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
    11641 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
    11701 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
    11761 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
    11821 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
    11881 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
    11941 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
    12001 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
    12061 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
    12121 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
    12181 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
    12241 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
    12301 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
    12361 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
    12421 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
    12481 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
    12541 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
    12601 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
    12661 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
    12721 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
    12781 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
    12841 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
    12901 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
    12961 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
    13021 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
    13081 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
    13141 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
    13201 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
    13261 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
    13321 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
    13381 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
    13441 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
    13501 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
    13561 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
    13621 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
    13681 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
    13741 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
    13801 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
    13861 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
    13921 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
    13981 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
    14041 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
    14101 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
    14161 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
    14221 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
    14281 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
    14341 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta g
//
