LOCUS       sequence-352527-       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_157
FEATURES             Location/Qualifiers
     source          1..12797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    85..106
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(121..126,127..144,145..151)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    159..175
                     /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     164..186
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     183..199
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     183..199
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     218..238
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        220..238
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        266..492
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             493..673
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    720..845
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1338..1571
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1756..1800
                     /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    2034..2151
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2290..3468
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2521..3459
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3416..3436
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3485..3505
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3512..5581,5582..5584,5585..5587,5588..5593,5594..5599,
                     5600..5602,5603..7612)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity without compromising on-target activity (Chen 
                     et al., 2017)"
                     /label=HypaCas9
                     /note="carries the mutations N692A, M694A, Q695A, and 
                     H698A"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRAFAA
                     LIADDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             3512..7612
                     /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
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRAFAA
                     LIADDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7619..7666
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7679..7732
                     /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             7739..8134
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             8135..8191
                     /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             8198..8896
                     /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    8915..9503
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8968..8988)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9386..9397)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9575..9808
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9886..10007
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9923..9942)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9977..9996
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10047..10182
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10202..10221)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10203..10221)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10231..10248)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10231..10247)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10240..10262)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10389..10844
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10476..10495)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10686..10707
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10870..10974
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10975..11043,11044..11835)
                     /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(11193..11212)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12006..12594
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12495..12514
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12748..12765
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
       61 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
      121 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
      181 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
      241 acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta gtcttgcaac
      301 atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg tgcatgccga
      361 ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca
      421 tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa gtgcctagct
      481 cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
      541 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
      601 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
      661 aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc
      721 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
      781 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
      841 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
      901 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
      961 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
     1021 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
     1081 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     1141 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga
     1201 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     1261 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     1321 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     1381 atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     1441 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     1501 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
     1561 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
     1621 tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg
     1681 gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg
     1741 caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg
     1801 tggaattggt ttaacataac aaattggctg tggtatataa aattattcat aatgatagta
     1861 ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg
     1921 cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg accctggtac
     1981 ccatgcatgc atgaattcta gatcttgaga caaatggcag tattcatcca caattttaaa
     2041 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
     2101 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat
     2161 tacagggaca gcagagatcc actttggcgc cggctcgagg gggcccgggt gcaaagatgg
     2221 ataaagtttt aaacagagag gaatctttgc agctaatgga ccttctaggt cttgaaagga
     2281 gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
     2341 aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac
     2401 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
     2461 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
     2521 gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg
     2581 ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga
     2641 agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt
     2701 gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt
     2761 ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt
     2821 tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt
     2881 tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg
     2941 ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct
     3001 ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg
     3061 tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca
     3121 aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg
     3181 gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg
     3241 cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt
     3301 tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac
     3361 ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag
     3421 cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgagg gatccaccat
     3481 ggcaccaaag aagaagcgga aggtcgcagc cgacaagaag tacagcatcg gcctggacat
     3541 cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa
     3601 attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct
     3661 gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag
     3721 atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc
     3781 caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa
     3841 gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa
     3901 gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct
     3961 gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga
     4021 gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac
     4081 ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtagacg ccaaggccat
     4141 cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg
     4201 cgagaagaag aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa
     4261 cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta
     4321 cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct
     4381 ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga
     4441 gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga
     4501 cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt
     4561 cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga
     4621 gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt
     4681 gaagctgaac agagaggacc tgctgcggaa gcagcgtacg ttcgacaacg gcagcatccc
     4741 ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc
     4801 attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta
     4861 cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga
     4921 aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgctagcg cccagagctt
     4981 catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca
     5041 cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac
     5101 cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct
     5161 gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa
     5221 aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct
     5281 gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga
     5341 aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat
     5401 gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct
     5461 gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg
     5521 ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag
     5581 agccttcgcc gccctgatcg ccgacgacag cctgaccttt aaagaggaca tccagaaagc
     5641 ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc
     5701 cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat
     5761 gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca
     5821 gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct
     5881 gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct
     5941 gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa
     6001 ccggctgtcc gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc
     6061 catcgacaac aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc
     6121 ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct
     6181 gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact
     6241 ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt
     6301 ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg
     6361 ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca
     6421 gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc
     6481 cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg
     6541 cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa
     6601 ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac
     6661 cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga
     6721 gatcgtgtgg gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca
     6781 agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct
     6841 gcccaagagg aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta
     6901 cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa
     6961 gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag
     7021 aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa
     7081 aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa
     7141 gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa
     7201 atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga
     7261 taatgagcag aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga
     7321 gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct
     7381 gtccgcctac aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca
     7441 cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat
     7501 cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag
     7561 catcaccggc ctgtacgaga cacgaatcga tttgtctcag ctgggaggcg acgccgcaaa
     7621 aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag aaaaaggcag gctcaggcga
     7681 gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc caagcgctat
     7741 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
     7801 cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg
     7861 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
     7921 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
     7981 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
     8041 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
     8101 tgaattgctg ccctctggtt atgtgtggga gggcgcaaca aacttctctc tgctgaaaca
     8161 agccggagat gtcgaagaga atcctggacc gactagtatg gtgagcgagc tgattaagga
     8221 gaacatgcac atgaagctat acatggaggg caccgtgaac aaccaccact tcaagtgcac
     8281 atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca aggcggtcga
     8341 gggcggccct ctccccttcg ccttcgacat cctggctacc agcttcatgt acggcagcaa
     8401 aaccttcatc aaccacaccc agggcatccc cgacttcttt aagcagtcct tccccgaggg
     8461 cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga
     8521 caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttccc
     8581 atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggcctcca ccgagaccct
     8641 gtaccccgct gacggcggcc tggaaggcag agccgacatg gccctgaagc tcgtgggcgg
     8701 gggccacctg atctgcaact tgaagaccac atacagatcc aagaaacccg ctaagaacct
     8761 caagatgccc ggcgtctact atgtggacag aagactggaa agaatcaagg aggccgacaa
     8821 agagacctac gtcgagcagc acgaggtggc tgtggccaga tactgcgacc tccctagcaa
     8881 actggggcac agatgataga cgcgttaagt cgacaatcaa cctctggatt acaaaatttg
     8941 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     9001 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     9061 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     9121 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     9181 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     9241 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     9301 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     9361 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     9421 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     9481 ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta
     9541 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     9601 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     9661 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     9721 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     9781 ttttagtcag tgtggaaaat ctctagcagt acgtatagta gttcatgtca tcttattatt
     9841 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     9901 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     9961 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
    10021 tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
    10081 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
    10141 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc
    10201 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
    10261 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
    10321 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
    10381 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
    10441 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
    10501 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
    10561 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
    10621 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
    10681 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
    10741 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
    10801 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
    10861 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
    10921 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
    10981 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
    11041 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
    11101 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
    11161 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
    11221 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
    11281 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
    11341 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
    11401 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
    11461 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
    11521 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
    11581 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
    11641 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
    11701 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
    11761 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
    11821 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
    11881 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
    11941 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
    12001 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
    12061 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
    12121 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
    12181 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
    12241 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
    12301 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
    12361 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
    12421 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
    12481 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
    12541 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
    12601 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
    12661 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
    12721 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
    12781 ccaatacgca aaccgcc
//
