LOCUS       sequence-77408-e        9318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9 nickase optimized for nuclear import..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9318)
  AUTHORS   Shen B, Zhang W, Zhang J, Zhou J, Wang J, Chen L, Wang L, Hodgkins 
            A, Iyer V, Huang X, Skarnes WC
  TITLE     Efficient genome modification by CRISPR-Cas9 nickase with minimal 
            off-target effects.
  JOURNAL   Nat Methods. 2014 Mar 2. doi: 10.1038/nmeth.2857.
  PUBMED    24584192
REFERENCE   2  (bases 1 to 9318)
  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 
            Methods. 2014 Mar 2. doi: 10.1038/nmeth.2857."
COMMENT     Sequence Label: pST1374-N-NLS-flag-linker-Cas9-H840A
FEATURES             Location/Qualifiers
     source          1..9318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             910..930
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             940..963
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(1066..3579,3580..3582,3583..5169)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the H840A mutation in the HNH catalytic domain"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5182..5223
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5233..5250
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5273..5290)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5279..5503
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5549..5977
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5636..5655)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5846..5867
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5986..6006)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5991..6321
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6172..6307
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6234..6253
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6369..6416
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6435..6833
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    6991..7112
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7028..7047)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7082..7101
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7161..7177)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7161..7177)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7174..7196)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7185..7201
                     /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(7209..7215,7216..7233,7234..7239))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7254..7275
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7392..7409)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7563..8151)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7643..7662)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8322..9113,9114..9182))
                     /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     8945..8964
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9183..9287)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 accatgggac ctaagaaaaa gaggaaggtg gcggccgctg actacaagga tgacgacgat
      961 aaatctagac tggaacccgg cgagaagcca tacaaatgcc cagagtgtgg taaatctttc
     1021 tctcagtctg gtgctctgac cagacaccag cggacccaca ctagagacaa gaaatactct
     1081 attggactgg atatcgggac aaactccgtt ggctgggccg tcataaccga cgagtataag
     1141 gtgccaagca agaaattcaa ggtgctgggt aatactgacc gccattcaat caagaagaac
     1201 ctgatcggag cactcctctt cgactccggt gaaaccgctg aagctactcg gctgaagcgg
     1261 accgcaaggc ggagatacac ccgccgcaag aatcggatat gttatctgca agagatcttt
     1321 agcaacgaaa tggctaaggt ggacgactcc ttctttcacc gcctggaaga gagctttctg
     1381 gtggaggagg ataagaaaca cgagaggcac cctatattcg gaaatatcgt ggatgaggtg
     1441 gcttaccatg aaaagtatcc tacaatctac catctgagga agaagctggt ggacagcacc
     1501 gataaagcag acctgaggct catctatctg gccctggctc atatgataaa gtttagagga
     1561 cactttctga tcgagggcga cctgaatccc gataattccg atgtggataa actcttcatt
     1621 caactggtgc agacatataa ccaactgttc gaggagaatc ccataaacgc ttctggtgtg
     1681 gatgccaagg ctattctgtc cgctcggctg tccaagtcac gcagactgga gaatctgatt
     1741 gcccaactgc caggagaaaa gaagaacggc ctgtttggga acctcatcgc cctgagcctg
     1801 ggcctgacac ctaacttcaa gtccaatttt gatctggccg aagatgctaa actccagctc
     1861 tccaaggaca cctatgacga tgatctggac aacctgctcg cacagatagg cgaccagtac
     1921 gccgatctct ttctggctgc taagaatctc tccgacgcca ttctgctgag cgacatactc
     1981 cgggtcaaca ctgagatcac caaagcacct ctgagcgcct ccatgataaa acgctatgat
     2041 gaacaccatc aagacctgac tctgctcaaa gccctcgtga ggcaacagct gccagagaag
     2101 tacaaagaga tattcttcga ccagagcaag aatggatatg ccggatacat cgatggcgga
     2161 gcatcacagg aagaatttta caagttcatc aaaccaatcc tcgagaagat ggacggtact
     2221 gaagagctgc tggtgaagct gaacagggag gacctgctga ggaagcagag gacctttgat
     2281 aatggctcca ttccacatca gatacacctg ggagagctgc atgcaatcct ccgcaggcag
     2341 gaggatttct atcctttcct gaaggataac cgggagaaga tagagaagat cctgaccttc
     2401 aggatccctt attacgtcgg ccctctggct agaggcaact cccgcttcgc ttggatgacc
     2461 aggaaatctg aggagacaat tactccttgg aacttcgaag aggtcgtgga taagggcgca
     2521 agcgcccagt cattcatcga acggatgacc aatttcgata agaacctgcc caacgagaag
     2581 gtcctgccca aacattcact cctgtacgag tatttcaccg tctataacga gctgactaaa
     2641 gtgaagtacg tgaccgaggg catgaggaag cctgccttcc tgtccggaga gcagaagaag
     2701 gctatcgttg atctgctctt caagactaat agaaaggtga cagtgaagca gctcaaggag
     2761 gattacttta agaagatcga atgctttgac tcagtggaaa tctctggcgt ggaggaccgc
     2821 tttaatgcca gcctgggcac ttaccatgat ctgctgaaga taatcaaaga caaagatttc
     2881 ctcgataatg aggagaacga ggacatcctg gaagatatcg tgctgaccct gactctgttc
     2941 gaggatagag agatgatcga agagcgcctg aagacctatg cccatctgtt tgacgataaa
     3001 gtcatgaaac agctcaagcg gcggcgctac actgggtggg gtagactctc caggaaactc
     3061 ataaacggca tccgcgacaa acagagcgga aagaccatcc tggatttcct gaaatccgac
     3121 ggattcgcta acaggaactt catgcaactg attcacgatg actctctgac atttaaagag
     3181 gacatccaga aggcacaggt gagcggtcaa ggcgacagcc tgcacgagca catcgccaac
     3241 ctcgctggat cacccgccat aaagaaggga atactgcaga cagtcaaggt cgtggacgaa
     3301 ctcgtcaaag tgatgggtcg gcacaagcca gagaatatcg ttatcgaaat ggcaagggag
     3361 aaccaaacca cccagaaggg ccagaagaac tctcgggaac ggatgaaaag aatcgaagag
     3421 ggaattaagg agctgggatc tcagatactg aaggagcacc ctgtggagaa tacacagctc
     3481 cagaacgaga aactctacct gtactacctc cagaacgggc gggacatgta cgttgaccag
     3541 gaactcgaca tcaaccggct gtccgattat gacgtggacg ctattgttcc acagtccttc
     3601 ctcaaagatg actccattga caacaaggtg ctgaccagat ccgataagaa tcgcggtaag
     3661 tctgacaatg ttccatcaga agaggtggtc aagaagatga agaattactg gcggcagctc
     3721 ctcaacgcca aactgatcac ccagcggaag tttgacaatc tgactaaggc agaaagagga
     3781 ggtctgagcg aactcgacaa ggccggcttt attaagaggc aactggtcga aacacgccag
     3841 attaccaaac acgtggcaca aatcctcgac tctaggatga acactaagta cgatgagaac
     3901 gataagctga tcagggaagt gaaagtgata actctgaaga gcaagctggt gtctgacttc
     3961 cggaaggact ttcaattcta caaagttcgc gaaataaaca attaccatca tgctcacgat
     4021 gcctatctca atgctgtcgt tggcaccgcc ctgatcaaga aataccctaa actggagtct
     4081 gagttcgtgt acggtgacta taaagtctac gatgtgagga agatgatagc aaagtctgag
     4141 caagagattg gcaaagccac cgccaagtac ttcttctact ctaatatcat gaatttcttt
     4201 aagactgaga taaccctggc taacggcgaa atccggaagc gcccactgat cgaaacaaac
     4261 ggagaaacag gagaaatcgt gtgggataaa ggcagggact tcgcaactgt gcggaaggtg
     4321 ctgtccatgc cacaagtcaa tatcgtgaag aagaccgaag tgcagaccgg cggattctca
     4381 aaggagagca tcctgccaaa gcggaactct gacaagctga tcgccaggaa gaaagattgg
     4441 gacccaaaga agtatggcgg tttcgattcc cctacagtgg cttattccgt tctggtcgtg
     4501 gcaaaagtgg agaaaggcaa gtccaagaaa ctcaagtctg ttaaggagct gctcggaatt
     4561 actattatgg agagatccag cttcgagaag aatccaatcg atttcctgga agctaagggc
     4621 tataaagaag tgaagaaaga tctcatcatc aaactgccca agtactctct ctttgagctg
     4681 gagaatggta ggaagcggat gctggcctcc gccggagagc tgcagaaagg aaacgagctg
     4741 gctctgccct ccaaatacgt gaacttcctg tatctggcct cccactacga gaaactcaaa
     4801 ggtagccctg aagacaatga gcagaagcaa ctctttgttg agcaacataa acactacctg
     4861 gacgaaatca ttgaacagat tagcgagttc agcaagcggg ttattctggc cgatgcaaac
     4921 ctcgataaag tgctgagcgc atataataag cacagggaca agccaattcg cgaacaagca
     4981 gagaatatta tccacctctt tactctgact aatctgggcg ctcctgctgc cttcaagtat
     5041 ttcgatacaa ctattgacag gaagcggtac acctctacca aagaagttct cgatgccacc
     5101 ctgatacacc agtcaattac cggactgtac gagactcgca tcgacctgtc tcagctcggc
     5161 ggcgactagg ggcccttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct
     5221 acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact
     5281 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     5341 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     5401 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     5461 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     5521 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     5581 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     5641 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5701 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5761 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5821 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5881 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta
     5941 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     6001 tagggtgtgg aaagtcccca ggctccccag gcaggcagaa gtatgcaaag catgcatctc
     6061 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     6121 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     6181 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     6241 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     6301 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     6361 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     6421 tgaggaacta aaccatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag
     6481 caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc
     6541 tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt
     6601 gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta
     6661 tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgtcgacagg
     6721 tgcttctcga tctgcatcct gggatcaaag cgatagtgaa ggacagtgat ggacagccga
     6781 cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taagcacttc
     6841 gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat
     6901 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     6961 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     7021 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     7081 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     7141 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     7201 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     7261 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     7321 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     7381 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     7441 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     7501 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     7561 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     7621 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     7681 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     7741 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     7801 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     7861 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     7921 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7981 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     8041 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     8101 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     8161 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     8221 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     8281 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     8341 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     8401 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     8461 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     8521 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     8581 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     8641 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     8701 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     8761 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     8821 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     8881 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     8941 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     9001 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     9061 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     9121 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     9181 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     9241 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     9301 aaagtgccac ctgacgtc
//
