LOCUS       sequence-388903-       14235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized SpCas9-NRCH-YE1-BE4max and 
            blasticidin resistance: EFS 
            promoter-SpCas9-NRCH-YE1-BE4max-NLS-FLAG-P2A-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14235)
  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 14235)
  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-YE1-BE4max
FEATURES             Location/Qualifiers
     source          1..14235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        114..493
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        495..693
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     648..668
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             711..891
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    938..1063
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1556..1789
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1974..2018
                     /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    2316..2433
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2526..2737
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2788..2808
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3589..7689
                     /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             7720..7968
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7999..8247
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             8290..8310
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8311..8334
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8344..8400
                     /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             8401..8799
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8824..9412
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8877..8897)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9295..9306)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9415..9431)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9416..9432)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9937..10117
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10143..10160)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10149..10373
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10419..10847
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10506..10525)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10716..10737
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10856..10876)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      11021..11155
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11082..11101
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11217..11264
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11283..11657
                     /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    11787..11908
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11824..11843)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11878..11897
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11957..11973)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11957..11973)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11970..11992)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11981..11997
                     /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(12005..12011,12012..12029,12030..12035))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12050..12071
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12188..12205)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12359..12947)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12439..12458)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13118..13909,13910..13978))
                     /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     13741..13760
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13979..14083)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14158..14177)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
       61 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
      121 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      181 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      241 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      301 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      361 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      421 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      481 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      541 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      601 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      661 taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact gggtctctct
      721 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
      781 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
      841 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
      901 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
      961 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
     1021 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
     1081 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
     1141 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
     1201 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
     1261 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
     1321 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
     1381 taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg
     1441 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1501 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1561 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     1621 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1681 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1741 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1801 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1861 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     1921 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     1981 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     2041 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     2101 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     2161 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
     2221 gtggagagag agacagagac agatccattc gattagtgaa cggatcggca ctgcgtgcgc
     2281 caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     2341 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     2401 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     2461 ccagtttggt taattagcta gctaggtctt gaaaggagtg ggaattggct ccggtgcccg
     2521 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     2581 ttgatccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     2641 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     2701 cgttcttttt cgcaacgggt ttgccgccag aacacaggac cggttgccac catgaaacgg
     2761 acagccgacg gaagcgagtt cgagtcacca aagaagaagc ggaaagtctc ctcagagact
     2821 gggcctgtcg ccgtcgatcc aaccctgcgc cgccggattg aacctcacga gtttgaagtg
     2881 ttctttgacc cccgggagct gagaaaggag acatgcctgc tgtacgagat caactgggga
     2941 ggcaggcact ccatctggag gcacacctct cagaacacaa ataagcacgt ggaggtgaac
     3001 ttcatcgaga agtttaccac agagcggtac ttctgcccca ataccagatg tagcatcaca
     3061 tggtttctga gctattcccc ttgcggagag tgtagcaggg ccatcaccga gttcctgtcc
     3121 agatatccac acgtgacact gtttatctac atcgccaggc tgtatcacca cgcagaccca
     3181 gagaataggc agggcctgcg cgatctgatc agctccggcg tgaccatcca gatcatgaca
     3241 gagcaggagt ccggctactg ctggcggaac ttcgtgaatt attctcctag caacgaggcc
     3301 cactggccta ggtacccaca cctgtgggtg cgcctgtacg tgctggagct gtattgcatc
     3361 atcctgggcc tgcccccttg tctgaatatc ctgcggagaa agcagcccca gctgaccttc
     3421 tttacaatcg ccctgcagtc ttgtcactat cagaggctgc caccccacat cctgtgggcc
     3481 acaggcctga agtctggagg atctagcgga ggatcctctg gcagcgagac accaggaaca
     3541 agcgagtcag caacaccaga gagcagtggc ggcagcagcg gcggcagcga caagaagtac
     3601 agcatcggcc tggccatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac
     3661 aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag
     3721 aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag
     3781 agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc
     3841 ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc
     3901 ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag
     3961 gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc
     4021 accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg
     4081 ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc
     4141 atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc
     4201 gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg
     4261 atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc
     4321 ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag
     4381 ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag
     4441 tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc
     4501 ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatggt gaagagatac
     4561 gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag
     4621 aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc
     4681 ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc
     4741 accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc
     4801 gacaacggca tcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg
     4861 cagggcgatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc
     4921 ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg
     4981 accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc
     5041 gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag
     5101 aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc
     5161 aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa
     5221 aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa
     5281 gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat
     5341 cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac
     5401 ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg
     5461 tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac
     5521 aaagtgatga agcagctgaa gcggctgaga tacaccggct ggggcaggct gagccggaag
     5581 ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc
     5641 gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa
     5701 gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc
     5761 aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac
     5821 gagctcgtga aagtgatggg cggccacaag cccgagaaca tcgtgatcga aatggccaga
     5881 gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa
     5941 gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag
     6001 ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac
     6061 caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc
     6121 tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc
     6181 aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag
     6241 ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga
     6301 ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg
     6361 cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag
     6421 aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat
     6481 ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac
     6541 gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa
     6601 agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc
     6661 gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt
     6721 ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca
     6781 aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa
     6841 gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc
     6901 agcaaagagt ctatcctgcc caagggcaac agcgataagc tgatcgccag aaagaaggac
     6961 tgggacccta agaagtacgg cggcttcaac agccccaccg tggcctattc tgtgctggtg
     7021 gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctactgggg
     7081 atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag
     7141 ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag
     7201 ctggaaaacg gccggaagag aatgctggcc tctgccggcg tgctgcagaa gggaaacgaa
     7261 ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg
     7321 aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac
     7381 ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct
     7441 aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag
     7501 gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag
     7561 tactttgaca ccaccatcaa ccggaagcag tacaacacca ccaaagaggt gctggacgcc
     7621 accctgatcc ggcagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg
     7681 ggaggcgaca gcggcgggag cggcgggagc ggggggagca ctaatctgag cgacatcatt
     7741 gagaaggaga ctgggaaaca gctggtcatt caggagtcca tcctgatgct gcctgaggag
     7801 gtggaggaag tgatcggcaa caagccagag tctgacatcc tggtgcacac cgcctacgac
     7861 gagtccacag atgagaatgt gatgctgctg acctctgacg cccccgagta taagccttgg
     7921 gccctggtca tccaggattc taacggcgag aataagatca agatgctgag cggaggatcc
     7981 ggaggatctg gaggcagcac caacctgtct gacatcatcg agaaggagac aggcaagcag
     8041 ctggtcatcc aggagagcat cctgatgctg cccgaagaag tcgaagaagt gatcggaaac
     8101 aagcctgaga gcgatatcct ggtccatacc gcctacgacg agagtaccga cgaaaatgtg
     8161 atgctgctga catccgacgc cccagagtat aagccctggg ctctggtcat ccaggattcc
     8221 aacggagaga acaaaatcaa aatgctgtct ggcggctcaa aaagaaccgc cgacggcagc
     8281 gaattcgagc ccaagaagaa gaggaaagtc gattacaaag acgatgacga taagggatcc
     8341 ggcgcaacaa acttctctct gctgaaacaa gccggagatg tcgaagagaa tcctggaccg
     8401 atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc
     8461 aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc
     8521 cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg
     8581 gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga
     8641 aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg
     8701 catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt
     8761 cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag aattcgatat caagcttatc
     8821 ggtaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     8881 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     8941 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     9001 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     9061 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     9121 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     9181 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     9241 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     9301 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     9361 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     9421 cgtcgacctc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     9481 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca
     9541 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     9601 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat
     9661 ctgtggatct accacacaca aggctacttc cctgattggc agaactacac accagggcca
     9721 gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag
     9781 aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat
     9841 gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt
     9901 catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc
     9961 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
    10021 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
    10081 ctcagaccct tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc
    10141 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
    10201 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
    10261 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
    10321 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
    10381 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
    10441 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
    10501 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
    10561 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
    10621 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
    10681 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
    10741 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
    10801 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
    10861 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
    10921 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
    10981 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
    11041 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
    11101 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
    11161 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg
    11221 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
    11281 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
    11341 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
    11401 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
    11461 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
    11521 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
    11581 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
    11641 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
    11701 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
    11761 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
    11821 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
    11881 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
    11941 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
    12001 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
    12061 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
    12121 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
    12181 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
    12241 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
    12301 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
    12361 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
    12421 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
    12481 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
    12541 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
    12601 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
    12661 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
    12721 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
    12781 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
    12841 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
    12901 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
    12961 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
    13021 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
    13081 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
    13141 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
    13201 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
    13261 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
    13321 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
    13381 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
    13441 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
    13501 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
    13561 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
    13621 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
    13681 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
    13741 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
    13801 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
    13861 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
    13921 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
    13981 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
    14041 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
    14101 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
    14161 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
    14221 cgctgagtag tgcgc
//
