LOCUS       sequence-388897-       12838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized SpCas9-NRRH and blasticidin 
            resistance: EFS promoter-SpCas9-NRRH-NLS-FLAG-P2A-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12838)
  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 12838)
  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-NRRH
FEATURES             Location/Qualifiers
     source          1..12838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        86..465
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        467..665
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     620..640
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             683..863
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    910..1035
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1528..1761
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1946..1990
                     /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    2288..2405
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2498..2709
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2728..2737
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2734..6837
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMVKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGIIPHQIHLGELHAILRRQGDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRLRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGG
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKGNSDKLIARKKDWDPKKYGGFNSPTAAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIGFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLHKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGVPAAFKYFDTTIDKKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6838..6885
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6886..6909
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6919..6975
                     /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             6976..7374
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    7399..7987
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7452..7472)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7870..7881)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7990..8006)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7991..8007)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8512..8692
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8718..8735)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8724..8948
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8994..9422
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9081..9100)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9291..9312
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9431..9451)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      9596..9730
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9657..9676
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9792..9839
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9858..10232
                     /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    10362..10483
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10399..10418)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10453..10472
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10532..10548)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10532..10548)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10545..10567)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10556..10572
                     /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(10580..10586,10587..10604,10605..10610))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10625..10646
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10763..10780)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10934..11522)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11014..11033)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11693..12484,12485..12553))
                     /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     12316..12335
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12554..12658)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12733..12752)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
       61 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
      121 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      181 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      241 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      301 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      361 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      421 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      481 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      541 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      601 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      661 agcagcgcgt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct
      721 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
      781 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
      841 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
      901 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
      961 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
     1021 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
     1081 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
     1141 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
     1201 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     1261 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     1321 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct
     1381 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata
     1441 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt
     1501 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc
     1561 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc
     1621 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt
     1681 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata
     1741 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac
     1801 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac
     1861 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat
     1921 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg
     1981 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat
     2041 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa
     2101 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg
     2161 acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat
     2221 tcgattagtg aacggatcgg cactgcgtgc gccaattctg cagacaaatg gcagtattca
     2281 tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag
     2341 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
     2401 attttcgggt ttattacagg gacagcagag atccagtttg gttaattagc tagctaggtc
     2461 ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca
     2521 gtccccgaga agttgggggg aggggtcggc aattgatccg gtgcctagag aaggtggcgc
     2581 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
     2641 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
     2701 agaacacagg accggttcta gagcgctgcc accatggaca agaagtacag catcggcctg
     2761 gacatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc
     2821 aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga
     2881 gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga
     2941 agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag
     3001 atggccaagg tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag
     3061 gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac
     3121 gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc
     3181 gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg
     3241 atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg
     3301 cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag
     3361 gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg
     3421 cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct gggcctgacc
     3481 cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac
     3541 acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg
     3601 tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac
     3661 accgagatca ccaaggcccc cctgagcgcc tctatggtga agagatacga cgagcaccac
     3721 caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag
     3781 attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag
     3841 gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg
     3901 ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcatc
     3961 atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca gggcgatttt
     4021 tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc
     4081 tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc
     4141 gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag
     4201 agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc
     4261 aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac
     4321 gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg
     4381 gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc
     4441 aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc
     4501 tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat
     4561 gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga
     4621 gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag
     4681 cagctgaagc ggctgagata caccggctgg ggcaggctga gccggaagct gatcaacggc
     4741 atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc
     4801 aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag
     4861 aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc
     4921 agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa
     4981 gtgatgggcg gccacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc
     5041 acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa
     5101 gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag
     5161 aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac
     5221 atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt tctgaaggac
     5281 gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac
     5341 gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc
     5401 aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc
     5461 gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag
     5521 cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg
     5581 atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat
     5641 ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg
     5701 aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg
     5761 tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc
     5821 ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag
     5881 attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc
     5941 ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg
     6001 ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct
     6061 atcctgccca agggcaacag cgataagctg atcgccagaa agaaggactg ggaccctaag
     6121 aagtacggcg gcttcaacag ccccaccgcc gcctattctg tgctggtggt ggccaaagtg
     6181 gaaaagggca agtccaagaa actgaagagt gtgaaagagc tactggggat caccatcatg
     6241 gaaagaagca gcttcgagaa gaatcccatc ggctttctgg aagccaaggg ctacaaagaa
     6301 gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc
     6361 cggaagagaa tgctggcctc tgccggcgtg ctgcacaagg gaaacgaact ggccctgccc
     6421 tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc
     6481 gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc
     6541 atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa
     6601 gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc
     6661 atccacctgt ttaccctgac caatctggga gtgcctgccg ccttcaagta ctttgacacc
     6721 accatcgaca agaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac
     6781 cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacaag
     6841 cgacctgccg ccacaaagaa ggctggacag gctaagaaga agaaagatta caaagacgat
     6901 gacgataagg gatccggcgc aacaaacttc tctctgctga aacaagccgg agatgtcgaa
     6961 gagaatcctg gaccgatggc caagcctttg tctcaagaag aatccaccct cattgaaaga
     7021 gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct
     7081 ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct
     7141 tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt
     7201 atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag
     7261 gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg
     7321 acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaagaattc
     7381 gatatcaagc ttatcggtaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     7441 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     7501 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     7561 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     7621 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     7681 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     7741 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     7801 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     7861 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     7921 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     7981 tccccgcatc gataccgtcg acctcgagac ctagaaaaac atggagcaat cacaagtagc
     8041 aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg
     8101 ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat
     8161 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     8221 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaac
     8281 tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta caagctagta
     8341 ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg cttgttacac
     8401 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg gaggtttgac
     8461 agccgcctag catttcatca catggcccga gagctgcatc cggactgtac tgggtctctc
     8521 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     8581 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     8641 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagggcccgt
     8701 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     8761 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     8821 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     8881 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     8941 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     9001 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     9061 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     9121 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     9181 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     9241 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     9301 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     9361 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     9421 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     9481 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     9541 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     9601 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     9661 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     9721 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     9781 gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     9841 aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg
     9901 acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg
     9961 aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg
    10021 accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt
    10081 acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga
    10141 ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact
    10201 gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg
    10261 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
    10321 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
    10381 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
    10441 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
    10501 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
    10561 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
    10621 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
    10681 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
    10741 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
    10801 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
    10861 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
    10921 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
    10981 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
    11041 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
    11101 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
    11161 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
    11221 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
    11281 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
    11341 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
    11401 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
    11461 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
    11521 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
    11581 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
    11641 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
    11701 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
    11761 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
    11821 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
    11881 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
    11941 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
    12001 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
    12061 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
    12121 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
    12181 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
    12241 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
    12301 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
    12361 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
    12421 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
    12481 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
    12541 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
    12601 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
    12661 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
    12721 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
    12781 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaa
//
