LOCUS       sequence-352529-       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_381
FEATURES             Location/Qualifiers
     source          1..12797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     132..151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     385..402
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    519..540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(555..560,561..578,579..585)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    593..609
                     /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     598..620
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     617..633
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     617..633
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     652..672
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        654..672
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        700..926
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             927..1107
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1154..1279
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1772..2005
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2190..2234
                     /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    2468..2585
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2724..3902
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2955..3893
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3850..3870
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3919..3939
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3946..8046
                     /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
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANANFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             8053..8100
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8113..8166
                     /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             8173..8568
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             8569..8625
                     /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             8632..9330
                     /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    9349..9937
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9402..9422)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9820..9831)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10009..10242
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10320..10441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10357..10376)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10411..10430
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10481..10616
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10636..10655)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10637..10655)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10665..10682)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10665..10681)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10674..10696)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10823..11278
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10910..10929)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11120..11141
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11304..11408
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11409..11477,11478..12269)
                     /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(11627..11646)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(12440..12797,1..231)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
       61 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      121 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      181 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      241 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      301 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      361 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      421 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      481 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
      541 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      601 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
      661 cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct
      721 tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc
      781 accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac
      841 agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat
      901 ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg
      961 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     1021 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     1081 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
     1141 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
     1201 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
     1261 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
     1321 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     1381 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     1441 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     1501 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     1561 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     1621 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     1681 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     1741 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     1801 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     1861 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     1921 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     1981 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     2041 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     2101 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     2161 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     2221 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     2281 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     2341 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     2401 ggggaccctg gtacccatgc atgcatgaat tctagatctt gagacaaatg gcagtattca
     2461 tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag
     2521 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
     2581 attttcgggt ttattacagg gacagcagag atccactttg gcgccggctc gagggggccc
     2641 gggtgcaaag atggataaag ttttaaacag agaggaatct ttgcagctaa tggaccttct
     2701 aggtcttgaa aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc
     2761 ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt
     2821 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
     2881 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
     2941 ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt
     3001 atggcccttg cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg
     3061 agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc
     3121 ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc
     3181 accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac
     3241 ctgctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca
     3301 ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat
     3361 gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag
     3421 ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg
     3481 caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg
     3541 ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca
     3601 cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc
     3661 gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt
     3721 ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta
     3781 ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt
     3841 ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt
     3901 gagggatcca ccatggcacc aaagaagaag cggaaggtcg cagccgacaa gaagtacagc
     3961 atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag
     4021 gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac
     4081 ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga
     4141 accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc
     4201 agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg
     4261 gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg
     4321 gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc
     4381 gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc
     4441 cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc
     4501 cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgta
     4561 gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc
     4621 gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg
     4681 ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg
     4741 agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac
     4801 gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg
     4861 agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac
     4921 gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag
     4981 tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga
     5041 gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc
     5101 gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg tacgttcgac
     5161 aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag
     5221 gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc
     5281 cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc
     5341 agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct
     5401 agcgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag
     5461 gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa
     5521 gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag
     5581 gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag
     5641 gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg
     5701 ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc
     5761 ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt
     5821 gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa
     5881 gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg
     5941 atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac
     6001 ggcttcgcca acgccaactt catgcagctg atccacgacg acagcctgac ctttaaagag
     6061 gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat
     6121 ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag
     6181 ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag
     6241 aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag
     6301 ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg
     6361 cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag
     6421 gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt
     6481 ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag
     6541 agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg
     6601 ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc
     6661 ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag
     6721 atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat
     6781 gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc
     6841 cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac
     6901 gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc
     6961 gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag
     7021 caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc
     7081 aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac
     7141 ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg
     7201 ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc
     7261 aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg
     7321 gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg
     7381 gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc
     7441 accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc
     7501 tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg
     7561 gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg
     7621 gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag
     7681 ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg
     7741 gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat
     7801 ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc
     7861 gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac
     7921 tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc
     7981 ctgatccacc agagcatcac cggcctgtac gagacacgaa tcgatttgtc tcagctggga
     8041 ggcgacgccg caaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag
     8101 gcaggctcag gcgagggcag aggaagtctg ctaacatgcg gtgacgtcga ggagaatcct
     8161 ggcccaagcg ctatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca
     8221 acggctacaa tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc
     8281 tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt
     8341 gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc
     8401 gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg
     8461 cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg
     8521 gcagttggga ttcgtgaatt gctgccctct ggttatgtgt gggagggcgc aacaaacttc
     8581 tctctgctga aacaagccgg agatgtcgaa gagaatcctg gaccgactag tatggtgagc
     8641 gagctgatta aggagaacat gcacatgaag ctatacatgg agggcaccgt gaacaaccac
     8701 cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga
     8761 atcaaggcgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc taccagcttc
     8821 atgtacggca gcaaaacctt catcaaccac acccagggca tccccgactt ctttaagcag
     8881 tccttccccg agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg
     8941 accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga
     9001 ggggtgaact tcccatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc
     9061 tccaccgaga ccctgtaccc cgctgacggc ggcctggaag gcagagccga catggccctg
     9121 aagctcgtgg gcgggggcca cctgatctgc aacttgaaga ccacatacag atccaagaaa
     9181 cccgctaaga acctcaagat gcccggcgtc tactatgtgg acagaagact ggaaagaatc
     9241 aaggaggccg acaaagagac ctacgtcgag cagcacgagg tggctgtggc cagatactgc
     9301 gacctcccta gcaaactggg gcacagatga tagacgcgtt aagtcgacaa tcaacctctg
     9361 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     9421 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     9481 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     9541 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     9601 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     9661 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     9721 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     9781 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     9841 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     9901 cagacgagtc ggatctccct ttgggccgcc tccccgcgtc gactttaaga ccaatgactt
     9961 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
    10021 ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
    10081 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
    10141 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
    10201 gatccctcag acccttttag tcagtgtgga aaatctctag cagtacgtat agtagttcat
    10261 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
    10321 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
    10381 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
    10441 atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc
    10501 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
    10561 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
    10621 gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
    10681 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
    10741 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
    10801 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
    10861 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
    10921 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
    10981 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
    11041 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
    11101 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
    11161 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
    11221 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
    11281 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
    11341 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
    11401 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
    11461 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
    11521 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
    11581 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
    11641 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
    11701 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
    11761 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
    11821 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
    11881 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
    11941 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
    12001 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
    12061 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
    12121 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
    12181 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
    12241 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
    12301 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
    12361 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
    12421 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
    12481 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
    12541 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
    12601 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
    12661 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
    12721 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
    12781 ccagcttgga gcgaacg
//
