LOCUS       sequence-352532-       14128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BE Cas expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14128)
  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 14128)
  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_336
FEATURES             Location/Qualifiers
     source          1..14128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    112..229
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        280..528
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     280..300
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     451..470
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        550..625
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        709..920
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             970..990
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             991..1674
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG
                     RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS
                     RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE
                     AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL
                     WATGLK"
     CDS             1771..5871
                     /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
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SIRPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARFLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPRAFKYFDTTIDRKVYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5902..6150
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6193..6213
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6223..6279
                     /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             6280..6876
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     6773..6793
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6892..7480
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6945..6965)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7363..7374)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7552..7785
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7811..7828)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7817..8041
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8087..8515
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8174..8193)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8384..8405
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8524..8544)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8529..8858
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8709..8844
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8771..8790
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8906..8953
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8972..9346
                     /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    9476..9597
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9513..9532)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9567..9586
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9646..9662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9646..9662)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9659..9681)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9670..9686
                     /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(9694..9700,9701..9718,9719..9724))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9739..9760
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9877..9894)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10048..10636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10128..10147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10807..11598,11599..11667))
                     /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     11430..11449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11668..11772)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(11847..11866)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        12038..12417
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        12419..12617
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     12572..12592
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             12635..12815
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    12862..12987
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    13480..13713
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             13898..13942
                     /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 ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga
       61 tcggcactgc gtgcgccaat tctgcagaca aatggcagta ttcatccaca attttaaaag
      121 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
      181 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
      241 cagggacagc agagatccag tttggttaat taaggtaccg agggcctatt tcccatgatt
      301 ccttcatatt tgcatatacg atacaaggct gttagagaga taattagaat taatttgact
      361 gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag
      421 tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag
      481 tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg agacgctttt
      541 ttcgtctcag tttgagagct agaaatagca agttcaaata aggctagtcc gttatcaact
      601 tgaaaaagtg gcaccgagtc ggtgcgctca cctattagcg gctaaggttt tttgaattcg
      661 ctagctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca
      721 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgatcc ggtgcctaga
      781 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
      841 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
      901 ggtttgccgc cagaacacag gaccggtgcc accatgaaac ggacagccga cggaagcgag
      961 ttcgagtcac caaagaagaa gcggaaagtc tcctcagaga ctgggcctgt cgccgtcgat
     1021 ccaaccctgc gccgccggat tgaacctcac gagtttgaag tgttctttga cccccgggag
     1081 ctgagaaagg agacatgcct gctgtacgag atcaactggg gaggcaggca ctccatctgg
     1141 aggcacacct ctcagaacac aaataagcac gtggaggtga acttcatcga gaagtttacc
     1201 acagagcggt acttctgccc caataccaga tgtagcatca catggtttct gagctggtcc
     1261 ccttgcggag agtgtagcag ggccatcacc gagttcctgt ccagatatcc acacgtgaca
     1321 ctgtttatct acatcgccag gctgtatcac cacgcagacc caaggaatag gcagggcctg
     1381 cgcgatctga tcagctccgg cgtgaccatc cagatcatga cagagcagga gtccggctac
     1441 tgctggcgga acttcgtgaa ttattctcct agcaacgagg cccactggcc taggtaccca
     1501 cacctgtggg tgcgcctgta cgtgctggag ctgtattgca tcatcctggg cctgccccct
     1561 tgtctgaata tcctgcggag aaagcagccc cagctgacct tctttacaat cgccctgcag
     1621 tcttgtcact atcagaggct gccaccccac atcctgtggg ccacaggcct gaagtctgga
     1681 ggatctagcg gaggatcctc tggcagcgag acaccaggaa caagcgagtc agcaacacca
     1741 gagagcagtg gcggcagcag cggcggcagc gacaagaagt acagcatcgg cctggccatc
     1801 ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa
     1861 ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg
     1921 ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga
     1981 tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc
     2041 aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag
     2101 aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag
     2161 taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg
     2221 cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag
     2281 ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc
     2341 tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc
     2401 ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc
     2461 gagaagaaga atggcctgtt cggaaacctg attgccctga gcctgggcct gacccccaac
     2521 ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac
     2581 gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg
     2641 gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag
     2701 atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac
     2761 ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc
     2821 ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag
     2881 ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg
     2941 aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc
     3001 caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca
     3061 ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac
     3121 gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa
     3181 accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc
     3241 atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac
     3301 agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc
     3361 gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg
     3421 ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa
     3481 atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg
     3541 ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa
     3601 aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg
     3661 atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg
     3721 aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg
     3781 gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga
     3841 aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc
     3901 caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc
     3961 gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg
     4021 ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag
     4081 aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg
     4141 ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg
     4201 tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac
     4261 cggctgtccg actacgatgt ggaccatatc gtgcctcaga gctttctgaa ggacgactcc
     4321 atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc
     4381 tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg
     4441 attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg
     4501 gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg
     4561 gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg
     4621 gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag
     4681 ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc
     4741 gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc
     4801 gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag
     4861 gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc
     4921 ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga aaccggggag
     4981 atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa
     5041 gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatccgg
     5101 cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac
     5161 ggcggcttcg tgagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag
     5221 ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga
     5281 agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa
     5341 aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag
     5401 agaatgctgg cctctgcccg gttcctgcag aagggaaacg aactggccct gccctccaaa
     5461 tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat
     5521 aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag
     5581 cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg
     5641 tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac
     5701 ctgtttaccc tgaccaatct gggagcccct cgggccttca agtactttga caccaccatc
     5761 gaccggaagg tgtaccggag caccaaagag gtgctggacg ccaccctgat ccaccagagc
     5821 atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggtga cagcggcggg
     5881 agcggcggga gcggggggag cactaatctg agcgacatca ttgagaagga gactgggaaa
     5941 cagctggtca ttcaggagtc catcctgatg ctgcctgagg aggtggagga agtgatcggc
     6001 aacaagccag agtctgacat cctggtgcac accgcctacg acgagtccac agatgagaat
     6061 gtgatgctgc tgacctctga cgcccccgag tataagcctt gggccctggt catccaggat
     6121 tctaacggcg agaataagat caagatgctg agcggaggct ccaaaagaac cgccgacggc
     6181 agcgagttcg agcccaagaa gaagaggaaa gtcagcgctg gcgcaacaaa cttctctctg
     6241 ctgaaacaag ccggagatgt cgaagagaat cctggaccga ccgagtacaa gcccacggtg
     6301 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     6361 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     6421 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     6481 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     6541 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     6601 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     6661 ggagtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     6721 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     6781 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     6841 cgcacctggt gcatgacccg caagcccggt gcctgaacgc gttaagtcga caatcaacct
     6901 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     6961 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     7021 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     7081 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     7141 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     7201 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     7261 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     7321 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     7381 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     7441 cctcagacga gtcggatctc cctttgggcc gcctccccgc gtcgacttta agaccaatga
     7501 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     7561 taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag
     7621 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     7681 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     7741 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc
     7801 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     7861 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     7921 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     7981 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     8041 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     8101 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     8161 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     8221 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     8281 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     8341 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     8401 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     8461 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     8521 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     8581 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     8641 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     8701 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     8761 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     8821 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     8881 ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat
     8941 agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt
     9001 gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc
     9061 ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat
     9121 cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg
     9181 cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc
     9241 cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga
     9301 cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga
     9361 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     9421 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     9481 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     9541 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     9601 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     9661 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     9721 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     9781 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     9841 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     9901 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     9961 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
    10021 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
    10081 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
    10141 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
    10201 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
    10261 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
    10321 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
    10381 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
    10441 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
    10501 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
    10561 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
    10621 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
    10681 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
    10741 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
    10801 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
    10861 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
    10921 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
    10981 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
    11041 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
    11101 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
    11161 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
    11221 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
    11281 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
    11341 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
    11401 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
    11461 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
    11521 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
    11581 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
    11641 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
    11701 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
    11761 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
    11821 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
    11881 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
    11941 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
    12001 gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat
    12061 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
    12121 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
    12181 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
    12241 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
    12301 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
    12361 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
    12421 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
    12481 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
    12541 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
    12601 gaggtctata taagcagcgc gttttgcctg tactgggtct ctctggttag accagatctg
    12661 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
    12721 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
    12781 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
    12841 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
    12901 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
    12961 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
    13021 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
    13081 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
    13141 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
    13201 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
    13261 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
    13321 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
    13381 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
    13441 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
    13501 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
    13561 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
    13621 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
    13681 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
    13741 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
    13801 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
    13861 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
    13921 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
    13981 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
    14041 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
    14101 aaccccgagg ggacccgaca ggcccgaa
//
