LOCUS       sequence-352530-       12797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  nuclease Cas expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12797)
  AUTHORS   Sangree AK, Griffith AL, Szegletes ZM, Roy P, DeWeirdt PC, Hegde M, 
            McGee AV, Hanna RE, Doench JG
  TITLE     Benchmarking of SpCas9 variants enables deeper base editor screens 
            of BRCA1 and BCL2.
  JOURNAL   Nat Commun. 2022 Mar 14;13(1):1318. doi: 10.1038/s41467-022-28884-7.
  PUBMED    35288574
REFERENCE   2  (bases 1 to 12797)
  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"; journalName: "Nat 
            Commun."; date: "2022-03-14"; volume: "13(1):1318. doi"; pages: " 
            10.1038/s41467-022-28884-7"
COMMENT     Sequence Label: pRDA_449
FEATURES             Location/Qualifiers
     source          1..12797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    105..222
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        361..1539
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          592..1530
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1487..1507
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1556..1576
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1583..5683
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFLWPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAKQLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5690..5737
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5750..5803
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5810..6205
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             6206..6262
                     /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             6269..6967
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     misc_feature    6986..7574
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7039..7059)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7457..7468)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7646..7879
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7957..8078
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7994..8013)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8048..8067
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8118..8253
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8273..8292)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8274..8292)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8302..8319)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8302..8318)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8311..8333)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8460..8915
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8547..8566)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8757..8778
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8941..9045
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9046..9114,9115..9906)
                     /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     complement(9264..9283)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10077..10665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10566..10585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10819..10836
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10953..10974
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10989..10994,10995..11012,11013..11019)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11027..11043
                     /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)."
     primer_bind     11032..11054
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11051..11067
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11051..11067
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11086..11106
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11088..11106
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        11134..11360
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             11361..11541
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11588..11713
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12206..12439
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12624..12668
                     /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 tcaccattat cgtttcagac ccacctccca accccgaggg gaccctggta cccatgcatg
       61 catgaattct agatcttgag acaaatggca gtattcatcc acaattttaa aagaaaaggg
      121 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
      181 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
      241 agcagagatc cactttggcg ccggctcgag ggggcccggg tgcaaagatg gataaagttt
      301 taaacagaga ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt
      361 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
      421 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      481 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      541 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      601 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      661 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      721 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      781 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      841 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      901 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      961 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
     1021 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
     1081 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1141 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1201 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1261 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1321 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1381 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1441 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1501 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag ggatccacca tggcaccaaa
     1561 gaagaagcgg aaggtcgcag ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa
     1621 ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt
     1681 gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga
     1741 cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag
     1801 acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga
     1861 cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga
     1921 gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac
     1981 catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat
     2041 ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct
     2101 gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca
     2161 gctgttcgag gaaaacccca tcaacgccag cggcgtagac gccaaggcca tcctgtctgc
     2221 cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa
     2281 gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag
     2341 caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga
     2401 cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa
     2461 gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa
     2521 ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct
     2581 gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca
     2641 gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa
     2701 gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa
     2761 cagagaggac ctgctgcgga agcagcgtac gttcgacaac ggcagcatcc cccaccagat
     2821 ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa
     2881 ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc
     2941 tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac
     3001 cccctggaac ttcgaggaag tggtggacaa gggcgctagc gcccagagct tcatcgagcg
     3061 gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct
     3121 gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat
     3181 gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa
     3241 gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg
     3301 cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata
     3361 ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga
     3421 cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga
     3481 acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg
     3541 gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca
     3601 gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat
     3661 gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc
     3721 cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa
     3781 gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca
     3841 caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca
     3901 gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca
     3961 gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta
     4021 ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc
     4081 cgactacgat gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgacaa
     4141 caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga
     4201 ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca
     4261 gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc
     4321 cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat
     4381 cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa
     4441 agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa
     4501 agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg
     4561 aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa
     4621 ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc
     4681 caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa
     4741 cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg
     4801 ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat
     4861 cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag
     4921 gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt
     4981 cctgtggccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc
     5041 caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt
     5101 cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct
     5161 gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct
     5221 ggcctctgcc aagcagctgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa
     5281 cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca
     5341 gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag
     5401 cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta
     5461 caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac
     5521 cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa
     5581 gcagtacaga agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg
     5641 cctgtacgag acacgaatcg atttgtctca gctgggaggc gacgccgcaa aaaggccggc
     5701 ggccacgaaa aaggccggcc aggcaaaaaa gaaaaaggca ggctcaggcg agggcagagg
     5761 aagtctgcta acatgcggtg acgtcgagga gaatcctggc ccaagcgcta tggccaagcc
     5821 tttgtctcaa gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc
     5881 catctctgaa gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac
     5941 tggtgtcaat gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac
     6001 tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag
     6061 gggcatcttg agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat
     6121 caaagccata gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct
     6181 gccctctggt tatgtgtggg agggcgcaac aaacttctct ctgctgaaac aagccggaga
     6241 tgtcgaagag aatcctggac cgactagtat ggtgagcgag ctgattaagg agaacatgca
     6301 catgaagcta tacatggagg gcaccgtgaa caaccaccac ttcaagtgca catccgaggg
     6361 cgaaggcaag ccctacgagg gcacccagac catgagaatc aaggcggtcg agggcggccc
     6421 tctccccttc gccttcgaca tcctggctac cagcttcatg tacggcagca aaaccttcat
     6481 caaccacacc cagggcatcc ccgacttctt taagcagtcc ttccccgagg gcttcacatg
     6541 ggagagagtc accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct
     6601 ccaggacggc tgcctcatct acaacgtcaa gatcagaggg gtgaacttcc catccaacgg
     6661 ccctgtgatg cagaagaaaa cactcggctg ggaggcctcc accgagaccc tgtaccccgc
     6721 tgacggcggc ctggaaggca gagccgacat ggccctgaag ctcgtgggcg ggggccacct
     6781 gatctgcaac ttgaagacca catacagatc caagaaaccc gctaagaacc tcaagatgcc
     6841 cggcgtctac tatgtggaca gaagactgga aagaatcaag gaggccgaca aagagaccta
     6901 cgtcgagcag cacgaggtgg ctgtggccag atactgcgac ctccctagca aactggggca
     6961 cagatgatag acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     7021 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     7081 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     7141 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     7201 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     7261 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     7321 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     7381 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     7441 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     7501 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     7561 ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc
     7621 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     7681 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     7741 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     7801 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     7861 gtgtggaaaa tctctagcag tacgtatagt agttcatgtc atcttattat tcagtattta
     7921 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     7981 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     8041 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     8101 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     8161 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     8221 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag
     8281 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     8341 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     8401 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     8461 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     8521 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     8581 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     8641 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     8701 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     8761 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     8821 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     8881 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     8941 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     9001 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     9061 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     9121 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     9181 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     9241 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     9301 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     9361 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     9421 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     9481 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     9541 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     9601 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     9661 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     9721 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     9781 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     9841 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     9901 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     9961 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
    10021 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
    10081 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
    10141 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
    10201 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
    10261 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
    10321 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
    10381 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
    10441 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
    10501 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
    10561 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
    10621 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
    10681 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
    10741 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
    10801 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
    10861 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
    10921 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
    10981 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
    11041 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
    11101 taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt
    11161 cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg
    11221 catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg
    11281 gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt
    11341 gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct
    11401 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
    11461 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
    11521 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
    11581 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
    11641 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
    11701 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
    11761 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
    11821 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
    11881 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
    11941 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
    12001 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga
    12061 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
    12121 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
    12181 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
    12241 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
    12301 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
    12361 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
    12421 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
    12481 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
    12541 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
    12601 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
    12661 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
    12721 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
    12781 gagttaggca gggatat
//
