LOCUS       sequence-352525-       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_155
FEATURES             Location/Qualifiers
     source          1..12797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    89..110
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(125..130,131..148,149..155)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    163..179
                     /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     168..190
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     187..203
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     187..203
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     222..242
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        224..242
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        270..496
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             497..677
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    724..849
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1342..1575
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1760..1804
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2038..2155
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2294..3472
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2525..3463
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3420..3440
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3489..3509
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3516..6914,6915..6917,6918..7514,7515..7517,7518..7520,
                     7521..7523,7524..7616)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             3516..7616
                     /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
                     SILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7623..7670
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7683..7736
                     /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             7743..8138
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             8139..8195
                     /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             8202..8900
                     /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    8919..9507
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8972..8992)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9390..9401)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9579..9812
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9890..10011
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9927..9946)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9981..10000
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10051..10186
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10206..10225)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10207..10225)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10235..10252)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10235..10251)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10244..10266)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10393..10848
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10480..10499)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10690..10711
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10874..10978
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10979..11047,11048..11839)
                     /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(11197..11216)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12010..12598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12499..12518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12752..12769
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
       61 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
      121 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
      181 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
      241 gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg
      301 caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg
      361 ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct
      421 gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct
      481 agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
      541 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
      601 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
      661 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag
      721 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
      781 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
      841 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg
      901 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa
      961 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga
     1021 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta
     1081 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac
     1141 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt
     1201 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
     1261 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
     1321 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
     1381 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
     1441 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
     1501 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
     1561 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
     1621 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
     1681 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
     1741 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
     1801 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
     1861 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt
     1921 taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga ggggaccctg
     1981 gtacccatgc atgcatgaat tctagatctt gagacaaatg gcagtattca tccacaattt
     2041 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     2101 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt
     2161 ttattacagg gacagcagag atccactttg gcgccggctc gagggggccc gggtgcaaag
     2221 atggataaag ttttaaacag agaggaatct ttgcagctaa tggaccttct aggtcttgaa
     2281 aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
     2341 cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt
     2401 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
     2461 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
     2521 acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg
     2581 cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg agcttcgggt
     2641 tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg
     2701 agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc
     2761 ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac
     2821 gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc
     2881 ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag
     2941 gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc
     3001 tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc
     3061 ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag
     3121 ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa
     3181 aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc
     3241 caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg
     3301 gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg
     3361 gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct
     3421 caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt gagggatcca
     3481 ccatggcacc aaagaagaag cggaaggtcg cagccgacaa gaagtacagc atcggcctgg
     3541 acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca
     3601 agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag
     3661 ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa
     3721 gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga
     3781 tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg
     3841 ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg
     3901 agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg
     3961 acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga
     4021 tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc
     4081 agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgta gacgccaagg
     4141 ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc
     4201 ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc
     4261 ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca
     4321 cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt
     4381 ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca
     4441 ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc
     4501 aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga
     4561 ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg
     4621 aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc
     4681 tcgtgaagct gaacagagag gacctgctgc ggaagcagcg tacgttcgac aacggcagca
     4741 tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt
     4801 acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct
     4861 actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg
     4921 aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct agcgcccaga
     4981 gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca
     5041 agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg
     5101 tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg
     5161 acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca
     5221 agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct
     5281 ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg
     5341 aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag
     5401 agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc
     5461 agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca
     5521 tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca
     5581 acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga
     5641 aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca
     5701 gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag
     5761 tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca
     5821 cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag
     5881 agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga
     5941 agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca
     6001 tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg
     6061 actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg
     6121 tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca
     6181 agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg
     6241 aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc
     6301 acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga
     6361 tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt
     6421 tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga
     6481 acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt
     6541 acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg
     6601 gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga
     6661 ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg
     6721 gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc
     6781 cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta
     6841 tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga
     6901 agtacggcgg cttcgtgagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg
     6961 aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg
     7021 aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag
     7081 tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc
     7141 ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct
     7201 ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg
     7261 aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca
     7321 tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag
     7381 tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca
     7441 tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca
     7501 ccatcgaccg gaagcagtac aggagcacca aagaggtgct ggacgccacc ctgatccacc
     7561 agagcatcac cggcctgtac gagacacgaa tcgatttgtc tcagctggga ggcgacgccg
     7621 caaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag gcaggctcag
     7681 gcgagggcag aggaagtctg ctaacatgcg gtgacgtcga ggagaatcct ggcccaagcg
     7741 ctatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa
     7801 tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg
     7861 gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg
     7921 tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg
     7981 gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc
     8041 tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga
     8101 ttcgtgaatt gctgccctct ggttatgtgt gggagggcgc aacaaacttc tctctgctga
     8161 aacaagccgg agatgtcgaa gagaatcctg gaccgactag tatggtgagc gagctgatta
     8221 aggagaacat gcacatgaag ctatacatgg agggcaccgt gaacaaccac cacttcaagt
     8281 gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga atcaaggcgg
     8341 tcgagggcgg ccctctcccc ttcgccttcg acatcctggc taccagcttc atgtacggca
     8401 gcaaaacctt catcaaccac acccagggca tccccgactt ctttaagcag tccttccccg
     8461 agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc
     8521 aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga ggggtgaact
     8581 tcccatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc tccaccgaga
     8641 ccctgtaccc cgctgacggc ggcctggaag gcagagccga catggccctg aagctcgtgg
     8701 gcgggggcca cctgatctgc aacttgaaga ccacatacag atccaagaaa cccgctaaga
     8761 acctcaagat gcccggcgtc tactatgtgg acagaagact ggaaagaatc aaggaggccg
     8821 acaaagagac ctacgtcgag cagcacgagg tggctgtggc cagatactgc gacctcccta
     8881 gcaaactggg gcacagatga tagacgcgtt aagtcgacaa tcaacctctg gattacaaaa
     8941 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     9001 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     9061 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     9121 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     9181 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     9241 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     9301 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     9361 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     9421 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     9481 ggatctccct ttgggccgcc tccccgcgtc gactttaaga ccaatgactt acaaggcagc
     9541 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     9601 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     9661 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     9721 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     9781 acccttttag tcagtgtgga aaatctctag cagtacgtat agtagttcat gtcatcttat
     9841 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat
     9901 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     9961 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
    10021 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
    10081 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
    10141 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg gacgtaccca
    10201 attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg
    10261 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
    10321 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
    10381 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
    10441 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
    10501 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
    10561 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
    10621 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
    10681 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
    10741 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
    10801 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
    10861 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
    10921 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
    10981 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
    11041 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
    11101 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
    11161 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
    11221 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
    11281 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
    11341 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
    11401 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
    11461 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
    11521 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
    11581 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
    11641 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
    11701 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
    11761 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
    11821 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
    11881 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
    11941 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
    12001 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
    12061 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
    12121 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
    12181 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
    12241 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
    12301 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
    12361 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
    12421 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
    12481 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
    12541 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
    12601 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
    12661 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
    12721 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
    12781 gcgcccaata cgcaaac
//
