LOCUS       sequence-388894-       14358 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized SpCas9-NRCH-SsAPOBEC3B and 
            blasticidin resistance: EFS 
            promoter-SpCas9-NRCH-SsAPOBEC3B-NLS-FLAG-P2A-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14358)
  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 14358)
  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-SsAPOBEC3B
FEATURES             Location/Qualifiers
     source          1..14358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        80..459
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        461..659
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     614..634
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             677..857
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    904..1029
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1522..1755
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1940..1984
                     /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    2282..2399
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2492..2703
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2712..2721
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3654..7754
                     /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             7785..8033
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             8064..8312
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             8313..8333
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8379..8399
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8400..8423
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8433..8489
                     /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             8490..8888
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8913..9501
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8966..8986)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9384..9395)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9504..9520)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9505..9521)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10026..10206
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10232..10249)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10238..10462
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10508..10936
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10595..10614)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10805..10826
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10945..10965)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      11110..11244
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11171..11190
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11306..11353
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11372..11746
                     /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    11876..11997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11913..11932)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11967..11986
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12046..12062)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12046..12062)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12059..12081)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12070..12086
                     /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(12094..12100,12101..12118,12119..12124))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12139..12160
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12277..12294)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12448..13036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12528..12547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13207..13998,13999..14067))
                     /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     13830..13849
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14068..14172)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14247..14266)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
       61 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
      121 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      181 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      241 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      301 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      361 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      421 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
      481 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      541 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
      601 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcagc
      661 gcgttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
      721 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
      781 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
      841 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag
      901 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
      961 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
     1021 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg
     1081 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa
     1141 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga
     1201 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta
     1261 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac
     1321 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt
     1381 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
     1441 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
     1501 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
     1561 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
     1621 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
     1681 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
     1741 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
     1801 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
     1861 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
     1921 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
     1981 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
     2041 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt
     2101 taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga
     2161 caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt
     2221 agtgaacgga tcggcactgc gtgcgccaat tctgcagaca aatggcagta ttcatccaca
     2281 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     2341 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc
     2401 gggtttatta cagggacagc agagatccag tttggttaat tagctagcta ggtcttgaaa
     2461 ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     2521 gagaagttgg ggggaggggt cggcaattga tccggtgcct agagaaggtg gcgcggggta
     2581 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     2641 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     2701 caggaccggt tgccaccatg gacccacaga ggctgcgcca gtggcccggc cctggcccag
     2761 caagcagggg cggctacggc cagcggccaa gaatcaggaa ccccgaggag tggtttcacg
     2821 agctgtctcc ccggaccttc agctttcact tccgcaacct gaggttcgca tccggccgca
     2881 atcggtctta tatctgctgt caggtggagg gcaagaactg cttctttcag ggcatctttc
     2941 agaatcaggt gccacctgac ccaccatgcc acgcagagct gtgcttcctg tcttggttcc
     3001 agagctgggg cctgtccccc gatgagcact actatgtgac atggtttatc tcttggagcc
     3061 cttgctgtga gtgtgccgcc aaggtggccc agttcctgga ggagaaccgc aacgtgagcc
     3121 tgtctctgag cgccgcaagg ctgtactatt tctggaagtc cgagtctaga gagggactgc
     3181 ggagactgag cgacctggga gcacaagtgg gaatcatgtc ctttcaggat ttccagcact
     3241 gctggaacaa ttttgtgcac aacctgggca tgcccttcca gccttggaag aagctgcaca
     3301 agaattacca gaggctggtg accgagctga agcagatcct gcgcgaggag cctgccacat
     3361 atggctctcc acaggcccag ggcaaggtga gaatcggaag caccgcagca ggactgaggc
     3421 acagccactc ccacacacgc tccgaggcac acctgaggcc taaccacagc tccagacagc
     3481 acaggatcct gaatcctcca cgggaggcca gagccaggac ctgcgtgctg gtggatgcct
     3541 cttggatctg ttacagaagc ggaggatcta gcggcggctc tagcggatct gagacacctg
     3601 gcacaagcga gtctgccaca cctgagagta gcggcggatc ttctggtggc tctgacaaga
     3661 agtacagcat cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg
     3721 agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca
     3781 agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc
     3841 tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag
     3901 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt
     3961 ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg
     4021 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg
     4081 acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt
     4141 tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc
     4201 tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca
     4261 gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa
     4321 atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc
     4381 tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac
     4441 tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg
     4501 accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg
     4561 acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct atggtgaaga
     4621 gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc
     4681 ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg
     4741 acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg
     4801 acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga
     4861 ccttcgacaa cggcatcatc ccccaccaga tccacctggg agagctgcac gccattctgc
     4921 ggcggcaggg cgatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc
     4981 tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct
     5041 ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca
     5101 agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca
     5161 acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc
     5221 tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc
     5281 agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc
     5341 tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg
     5401 aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca
     5461 aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga
     5521 cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg
     5581 acgacaaagt gatgaagcag ctgaagcggc tgagatacac cggctggggc aggctgagcc
     5641 ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga
     5701 agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct
     5761 ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca
     5821 ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg
     5881 tggacgagct cgtgaaagtg atgggcggcc acaagcccga gaacatcgtg atcgaaatgg
     5941 ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga
     6001 tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca
     6061 cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg
     6121 tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc
     6181 agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc
     6241 ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc
     6301 ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg
     6361 agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa
     6421 cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg
     6481 acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt
     6541 ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg
     6601 cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc
     6661 tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca
     6721 agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga
     6781 actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg
     6841 agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc
     6901 ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg
     6961 gcttcagcaa agagtctatc ctgcccaagg gcaacagcga taagctgatc gccagaaaga
     7021 aggactggga ccctaagaag tacggcggct tcaacagccc caccgtggcc tattctgtgc
     7081 tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctac
     7141 tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag
     7201 ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt
     7261 tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgtgctg cagaagggaa
     7321 acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga
     7381 agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc
     7441 actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg
     7501 acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag
     7561 agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct
     7621 tcaagtactt tgacaccacc atcaaccgga agcagtacaa caccaccaaa gaggtgctgg
     7681 acgccaccct gatccggcag agcatcaccg gcctgtacga gacacggatc gacctgtctc
     7741 agctgggagg cgactctggt ggaagcggag gatctggcgg cagcaccaat ctgagcgaca
     7801 tcatcgagaa agagacaggc aagcagctgg tcatccaaga gtccatcctg atgctgcctg
     7861 aagaggtgga agaagtgatc ggcaacaagc ccgagtccga catcctggtg cacaccgcct
     7921 acgatgagag caccgacgag aacgtgatgc tgctgacctc tgacgcccct gagtacaagc
     7981 cttgggctct cgtgatccag gacagcaacg gcgagaacaa gatcaagatg ctgagcggcg
     8041 gctctggtgg ctctggcgga tctacaaacc tgtccgatat tattgagaaa gaaaccggga
     8101 aacagctcgt gattcaagag tctattctca tgctcccgga agaagtcgag gaagtcattg
     8161 gaaacaagcc tgagagcgat attctggtcc atacagccta cgacgagtct accgatgaga
     8221 atgtcatgct cctcaccagc gacgctcccg agtataagcc atgggcactt gtcattcagg
     8281 actccaatgg ggaaaacaaa atcaaaatgc tcccaaagaa aaaacgcaag gtggagggag
     8341 ctgataagcg caccgccgat ggttccgagt tcgaaagccc caagaagaag aggaaagtcg
     8401 attacaaaga cgatgacgat aagggatccg gcgcaacaaa cttctctctg ctgaaacaag
     8461 ccggagatgt cgaagagaat cctggaccga tggccaagcc tttgtctcaa gaagaatcca
     8521 ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg
     8581 tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt
     8641 ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg
     8701 gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg
     8761 gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca
     8821 gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg
     8881 agggctaaga attcgatatc aagcttatcg gtaatcaacc tctggattac aaaatttgtg
     8941 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     9001 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     9061 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     9121 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     9181 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     9241 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     9301 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     9361 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     9421 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     9481 ccctttgggc cgcctccccg catcgatacc gtcgacctcg agacctagaa aaacatggag
     9541 caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag
     9601 aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca
     9661 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     9721 actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc
     9781 ctgattggca gaactacaca ccagggccag ggatcagata tccactgacc tttggatggt
     9841 gctacaagct agtaccagtt gagcaagaga aggtagaaga agccaatgaa ggagagaaca
     9901 cccgcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtattag
     9961 agtggaggtt tgacagccgc ctagcatttc atcacatggc ccgagagctg catccggact
    10021 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
    10081 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
    10141 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
    10201 ctagcagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
    10261 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
    10321 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
    10381 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
    10441 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
    10501 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
    10561 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
    10621 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
    10681 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
    10741 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
    10801 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
    10861 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
    10921 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
    10981 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
    11041 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
    11101 caaccatagt cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
    11161 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
    11221 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
    11281 ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat
    11341 agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt
    11401 gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc
    11461 ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat
    11521 cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg
    11581 cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc
    11641 cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga
    11701 cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga
    11761 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
    11821 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
    11881 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
    11941 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
    12001 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
    12061 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
    12121 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
    12181 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
    12241 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
    12301 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
    12361 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
    12421 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
    12481 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
    12541 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
    12601 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
    12661 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
    12721 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
    12781 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
    12841 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
    12901 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
    12961 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
    13021 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
    13081 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
    13141 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
    13201 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
    13261 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
    13321 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
    13381 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
    13441 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
    13501 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
    13561 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
    13621 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
    13681 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
    13741 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
    13801 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
    13861 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
    13921 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
    13981 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
    14041 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
    14101 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
    14161 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
    14221 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
    14281 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
    14341 acaacaaggc aaggcttg
//
