LOCUS       sequence-274793-       12838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized SpCas9-QQR1 and blasticidin 
            resistance: EFS promoter-QQR1-NLS-FLAG-P2A-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12838)
  AUTHORS   Kim N, Kim HK, Lee S, Seo JH, Choi JW, Park J, Min S, Yoon S, Cho 
            SR, Kim HH
  TITLE     Prediction of the sequence-specific cleavage activity of Cas9 
            variants.
  JOURNAL   Nat Biotechnol. 2020 Nov;38(11):1328-1336. doi: 
            10.1038/s41587-020-0537-9. Epub 2020 Jun 8.
  PUBMED    32514125
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: 
            "10.1038/s41587-020-0537-9"; journalName: "Nat Biotechnol"; date: 
            "2020-11"; volume: "38"; issue: "11"; pages: "1328-1336"
COMMENT     Sequence Label: QQR1 variant
FEATURES             Location/Qualifiers
     source          1..12838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    119..236
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        329..540
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      559..568
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             565..4668
                     /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
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     AQLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTFKQKQYRSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4669..4716
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4717..4740
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4750..4806
                     /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             4807..5205
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    5230..5818
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5283..5303)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5701..5712)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5821..5837)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5822..5838)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6343..6523
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6549..6566)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6555..6779
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6825..7253
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6912..6931)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7122..7143
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7262..7282)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7427..7561
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7488..7507
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7623..7670
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7689..8063
                     /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    8193..8314
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8230..8249)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8284..8303
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8363..8379)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8363..8379)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8376..8398)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8387..8403
                     /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(8411..8417,8418..8435,8436..8441))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8456..8477
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8594..8611)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8765..9353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8845..8864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9524..10315,10316..10384))
                     /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     10147..10166
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10385..10489)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10564..10583)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        10755..11134
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11136..11334
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11289..11309
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             11352..11532
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11579..11704
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12197..12430
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12615..12659
                     /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"
ORIGIN
        1 gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt
       61 gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt
      121 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
      181 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg
      241 tttattacag ggacagcaga gatccagttt ggttaattag ctagctaggt cttgaaagga
      301 gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
      361 aagttggggg gaggggtcgg caattgatcc ggtgcctaga gaaggtggcg cggggtaaac
      421 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
      481 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
      541 gaccggttct agagcgctgc caccatggac aagaagtaca gcatcggcct ggacatcggc
      601 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
      661 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg
      721 ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac
      781 accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag
      841 gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag
      901 cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
      961 cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg
     1021 ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc
     1081 gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac
     1141 aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg
     1201 tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag
     1261 aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc
     1321 aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac
     1381 gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc
     1441 gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc
     1501 accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg
     1561 accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc
     1621 gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc
     1681 tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag
     1741 ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac
     1801 cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc
     1861 ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg
     1921 ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc
     1981 atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc
     2041 gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc
     2101 ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag
     2161 ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg
     2221 ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc
     2281 gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc
     2341 acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac
     2401 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
     2461 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
     2521 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
     2581 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
     2641 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
     2701 gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc
     2761 attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc
     2821 cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag
     2881 ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc
     2941 agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac
     3001 ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg
     3061 ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc
     3121 gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc
     3181 gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt
     3241 acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat
     3301 aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca
     3361 cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa
     3421 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt
     3481 tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc
     3541 gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac
     3601 tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct
     3661 accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg
     3721 gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc
     3781 gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg
     3841 aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc
     3901 aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc
     3961 ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc
     4021 aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc
     4081 agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag
     4141 gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga
     4201 atgctggcct ctgcccggga actgcagaag ggaaacgaac tggccctgcc ctccaaatat
     4261 gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat
     4321 gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag
     4381 atcagcgagt tctccaagag agtgatcctg gccgacgctc agctggacaa agtgctgtcc
     4441 gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg
     4501 tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac caccttcaag
     4561 cagaagcagt accggagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc
     4621 accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa gcgacctgcc
     4681 gccacaaaga aggctggaca ggctaagaag aagaaagatt acaaagacga tgacgataag
     4741 ggatccggcg caacaaactt ctctctgctg aaacaagccg gagatgtcga agagaatcct
     4801 ggaccgatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct
     4861 acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc
     4921 gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa
     4981 ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg
     5041 atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc
     5101 gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt
     5161 gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaagaatt cgatatcaag
     5221 cttatcggta atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     5281 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     5341 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     5401 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     5461 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     5521 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     5581 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     5641 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     5701 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     5761 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     5821 cgataccgtc gacctcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca
     5881 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca
     5941 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     6001 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc
     6061 cttgatctgt ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca
     6121 gggccaggga tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag
     6181 caagagaagg tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc
     6241 ctgcatggga tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta
     6301 gcatttcatc acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac
     6361 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     6421 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     6481 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagggcccg tttaaacccg
     6541 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     6601 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     6661 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     6721 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     6781 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     6841 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     6901 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     6961 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     7021 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     7081 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     7141 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     7201 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     7261 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     7321 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     7381 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     7441 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     7501 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     7561 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
     7621 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     7681 actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg
     7741 gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact
     7801 tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg
     7861 tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt
     7921 ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg
     7981 gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact
     8041 tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct
     8101 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     8161 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     8221 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     8281 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     8341 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     8401 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     8461 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     8521 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     8581 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     8641 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     8701 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     8761 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     8821 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     8881 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     8941 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     9001 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     9061 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     9121 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     9181 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     9241 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     9301 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     9361 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     9421 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     9481 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     9541 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     9601 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     9661 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     9721 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     9781 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     9841 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     9901 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     9961 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
    10021 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
    10081 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
    10141 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
    10201 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
    10261 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
    10321 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
    10381 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
    10441 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
    10501 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
    10561 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
    10621 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
    10681 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
    10741 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
    10801 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
    10861 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
    10921 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
    10981 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
    11041 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
    11101 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
    11161 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
    11221 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
    11281 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt
    11341 ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
    11401 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
    11461 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
    11521 aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct
    11581 ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact
    11641 ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag
    11701 cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg
    11761 gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt
    11821 cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct
    11881 acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac
    11941 cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat
    12001 agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac
    12061 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
    12121 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
    12181 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
    12241 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
    12301 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
    12361 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
    12421 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
    12481 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
    12541 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
    12601 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
    12661 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
    12721 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
    12781 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacag
//
