LOCUS       sequence-350180-       13838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression SV40 and nucleoplasmin NLS SpCas9 with 3 high 
            affinity MS2 loops and fused to an N-terminal HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13838)
  AUTHORS   Hung ME, Leonard JN
  TITLE     A platform for actively loading cargo RNA to elucidate limiting 
            steps in EV-mediated delivery.
  JOURNAL   J Extracell Vesicles. 2016 May 13;5:31027. doi: 
            10.3402/jev.v5.31027. eCollection 2016.
  PUBMED    27189348
REFERENCE   2  (bases 1 to 13838)
  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: "J Extracell
            Vesicles."; date: "2016-05-13"; volume: "5:31027. doi"; pages: " 
            10.3402/jev.v5.31027. eCollection 2016"
COMMENT     Sequence Label: pMEH305 HA NLS SpCas9 3x high affinity MS2 loops
FEATURES             Location/Qualifiers
     source          1..13838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2063..2180
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(2245..2619)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     regulatory      2616..2625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2629..2647)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2639..2686)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     enhancer        2791..3094
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3095..3298
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3249..3269
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3295..3319
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3397..3417
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     CDS             3418..3444
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3463..3483
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3508..7608
                     /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
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVGQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7609..7656
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     misc_feature    7799..8387
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7852..7872)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8270..8281)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_RNA        8437..8455
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        8476..8494
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        8514..8532
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     LTR             8608..8841
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8942..8959)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8948..9172
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9218..9646
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9305..9324)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9515..9536
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9655..9675)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9660..9989
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9840..9975
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9902..9921
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             10038..11063
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             10038..11063
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    11193..11314
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11363..11379)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11363..11379)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11376..11398)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11387..11403
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(11411..11417,11418..11435,11436..11441))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11456..11477
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11594..11611)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11765..12353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11845..11864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12524..13315,13316..13384))
                     /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     13147..13166
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13385..13489)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    13537..13671
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
     2041 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     2101 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     2161 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag
     2221 taccgggccc gctctagtcc ggaatcagtc ctgctcctcg gccacgaagt gcacgcagtt
     2281 gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat
     2341 ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag
     2401 ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg
     2461 gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac
     2521 gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg
     2581 cgcggtgagc accggaacgg cactggtcaa cttggccatg gtggccctcc tatagtgagt
     2641 cgtattatac tatgccgata tactatgccg atgattaatt gtcaacacgt gctgcaggtc
     2701 cgaggttcta gacgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
     2761 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     2821 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     2881 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     2941 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     3001 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     3061 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     3121 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     3181 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     3241 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
     3301 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag
     3361 acaccgactc tactagagga tctgccacca gctagcgaga atctgtactt tcagatgtac
     3421 ccatacgatg tgccagatta cgctggtggt agtagcggtg ggccaaagaa gaagcggaag
     3481 gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcggcct ggacatcggc
     3541 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
     3601 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg
     3661 ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac
     3721 accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag
     3781 gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag
     3841 cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
     3901 cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg
     3961 ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc
     4021 gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac
     4081 aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg
     4141 tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag
     4201 aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc
     4261 aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac
     4321 gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc
     4381 gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc
     4441 accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg
     4501 accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc
     4561 gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc
     4621 tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag
     4681 ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac
     4741 cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc
     4801 ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg
     4861 ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc
     4921 atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc
     4981 gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc
     5041 ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag
     5101 ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg
     5161 ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc
     5221 gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc
     5281 acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac
     5341 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
     5401 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
     5461 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
     5521 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
     5581 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
     5641 gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc
     5701 attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc
     5761 cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag
     5821 ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc
     5881 agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac
     5941 ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg
     6001 ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc
     6061 gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc
     6121 gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt
     6181 acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat
     6241 aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca
     6301 cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa
     6361 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt
     6421 tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc
     6481 gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac
     6541 tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct
     6601 accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg
     6661 gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc
     6721 gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg
     6781 aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc
     6841 aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc
     6901 ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc
     6961 aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc
     7021 agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag
     7081 gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga
     7141 atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat
     7201 gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat
     7261 gagcagaaac agctgtttgt gggacagcac aagcactacc tggacgagat catcgagcag
     7321 atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc
     7381 gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg
     7441 tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac
     7501 cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc
     7561 accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa aaggccggcg
     7621 gccacgaaaa aggccggcca ggcaaaaaag aaaaagtaag gcgcgccagt ccagtgtggt
     7681 ggaattctgc agatatccag cacagtggcg gccgctcgag tctagagggc ccgtttgctc
     7741 cgagggccgc caccacctgt tcctgtacgg catggacgag ctgtacaagt aggtcgacaa
     7801 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     7861 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     7921 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     7981 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     8041 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     8101 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     8161 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     8221 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     8281 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     8341 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattcgagc
     8401 tcggtacctt cccgggttca ttagatccta aggtaaacat gaggatcacc catgtctgca
     8461 ggtcgactct agaaaacatg aggatcaccc atgtcttggc actggcgtcg taaacatgag
     8521 gatcacccat gtctgcggta cctttaagac caatgactta caaggcagct gtagatctta
     8581 gccacttttt aaaagaaaag aggggactgg aagggctaat tcactcccaa cgaagacaag
     8641 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     8701 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     8761 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     8821 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     8881 ttgcaaagaa atgaatatca gagagtgaga ggccttgaca ttgtttaaac ccgctgatca
     8941 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     9001 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     9061 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     9121 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     9181 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     9241 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     9301 cccgctcctt tcgctttctt cccttccttt ctcgtcacgt tcgccggctt tccccgtcaa
     9361 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     9421 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     9481 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     9541 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     9601 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     9661 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     9721 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     9781 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     9841 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     9901 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     9961 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
    10021 gatctgatca gcacgtgatg aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc
    10081 tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc
    10141 gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg
    10201 atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc
    10261 cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg
    10321 cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg
    10381 tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc
    10441 cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg
    10501 ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg
    10561 cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg
    10621 tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca
    10681 ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct
    10741 ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg
    10801 agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct
    10861 atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg
    10921 caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg
    10981 ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca
    11041 ctcgtccgag ggcaaaggaa tagcacgtgc tacgagattt cgattccacc gccgccttct
    11101 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
    11161 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
    11221 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
    11281 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
    11341 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
    11401 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
    11461 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
    11521 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
    11581 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
    11641 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
    11701 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
    11761 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
    11821 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
    11881 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
    11941 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
    12001 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
    12061 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
    12121 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
    12181 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
    12241 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
    12301 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
    12361 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
    12421 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
    12481 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
    12541 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
    12601 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
    12661 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
    12721 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
    12781 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
    12841 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
    12901 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
    12961 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
    13021 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
    13081 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
    13141 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
    13201 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
    13261 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
    13321 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
    13381 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
    13441 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
    13501 gaaaagtgcc acctgacgtc gacggatcgg gagatcaact tgtttattgc agcttataat
    13561 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
    13621 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat caactggata
    13681 actcaagcta accaaaatca tcccaaactt cccaccccat accctattac cactgccaat
    13741 tacctgtggt ttcatttact ctaaacctgt gattcctctg aattattttc attttaaaga
    13801 aattgtattt gttaaatatg tactacaaac ttagtagt
//
