LOCUS       in_vivo_NmeCas9+        8571 bp    DNA     circular SYN 11-APR-2025
DEFINITION  All-in-one plasmid. Contains expression cassette for NmeCas9 with N 
            and C NLS and HA tag at the C, plus cassette (for cloning of spacer 
            in BsmBI) for expressing sgRNA under the control of U6 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8571)
  AUTHORS   Amrani N, Gao XD, Liu P, Edraki A, Mir A, Ibraheim R, Gupta A, 
            Sasaki KE, Wu T, Donohoue PD, Settle AH, Lied AM, McGovern K, Fuller
            CK, Cameron P, Fazzio TG, Zhu LJ, Wolfe SA, Sontheimer EJ
  TITLE     NmeCas9 is an intrinsically high-fidelity genome-editing platform.
  JOURNAL   Genome Biol. 2018 Dec 5;19(1):214. doi: 10.1186/s13059-018-1591-1.
  PUBMED    30518407
REFERENCE   2  (bases 1 to 8571)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 27, 2025 from SnapGene 8.0.3
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genome 
            Biol."; date: "2018-12-5"; volume: "19(1):214. doi"; pages: " 
            10.1186/s13059-018-1591-1"
FEATURES             Location/Qualifiers
     source          1..8571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        complement(36..156)
                     /label=Nm tracrRNA
                     /note="trans-activating CRISPR RNA for the Neisseria 
                     meningitidis CRISPR/Cas9 system (Hou et al., 2013)"
     primer_bind     complement(132..180)
                     /label=引物 1
     misc_feature    157..180
                     /label=EZH guide
     primer_bind     181..201
                     /label=引物 2
     promoter        complement(183..431)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(241..260)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(411..431)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(488..507)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        657..1819
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          888..1810
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1767..1787
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1825..1834
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1837..1857
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1858..4788
                     /codon_start=1
                     /product="Cas9 endonuclease from the Neisseria meningitidis
                     Type II CRISPR/Cas system (Esvelt et al., 2013)"
                     /label=NmCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="AAFKPNSINYILGLAIGIASVGWAMVEIDEEENPIRLIDLGVRVF
                     ERAEVPKTGDSLAMARRLARSVRRLTRRRAHRLLRTRRLLKREGVLQAANFDENGLIKS
                     LPNTPWQLRAAALDRKLTPLEWSAVLLHLIKHRGYLSQRKNEGETADKELGALLKGVAG
                     NAHALQTGDFRTPAELALNKFEKESGHIRNQRSDYSHTFSRKDLQAELILLFEKQKEFG
                     NPHVSGGLKEGIETLLMTQRPALSGDAVQKMLGHCTFEPAEPKAAKNTYTAERFIWLTK
                     LNNLRILEQGSERPLTDTERATLMDEPYRKSKLTYAQARKLLGLEDTAFFKGLRYGKDN
                     AEASTLMEMKAYHAISRALEKEGLKDKKSPLNLSPELQDEIGTAFSLFKTDEDITGRLK
                     DRIQPEILEALLKHISFDKFVQISLKALRRIVPLMEQGKRYDEACAEIYGDHYGKKNTE
                     EKIYLPPIPADEIRNPVVLRALSQARKVINGVVRRYGSPARIHIETAREVGKSFKDRKE
                     IEKRQEENRKDREKAAAKFREYFPNFVGEPKSKDILKLRLYEQQHGKCLYSGKEINLGR
                     LNEKGYVEIDHALPFSRTWDDSFNNKVLVLGSENQNKGNQTPYEYFNGKDNSREWQEFK
                     ARVETSRFPRSKKQRILLQKFDEDGFKERNLNDTRYVNRFLCQFVADRMRLTGKGKKRV
                     FASNGQITNLLRGFWGLRKVRAENDRHHALDAVVVACSTVAMQQKITRFVRYKEMNAFD
                     GKTIDKETGEVLHQKTHFPQPWEFFAQEVMIRVFGKPDGKPEFEEADTLEKLRTLLAEK
                     LSSRPEAVHEYVTPLFVSRAPNRKMSGQGHMETVKSAKRLDEGVSVLRVPLTQLKLKDL
                     EKMVNREREPKLYEALKARLEAHKDDPAKAFAEPFYKYDKAGNRTQQVKAVRVEQVQKT
                     GVWVRNHNGIADNATMVRVHPNDYKYQIDELGKEIRPCRLKKRPPVR"
     primer_bind     1882..1920
                     /label=604_D14A_F
     primer_bind     complement(1882..1901)
                     /label=603_D16A_R
     misc_feature    1900..1902
                     /label=D16A
     primer_bind     complement(4685..4715)
                     /label=607_celldPI_R
     primer_bind     4697..4743
                     /label=608_celldPI_F
     misc_feature    4705..4719
                     /label=PI
     CDS             4789..4815
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4822..4848
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     primer_bind     complement(4864..4881)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4870..5094
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5259..6372
                     /label=mini-oriP
                     /note="internally truncated variant of the Epstein-Barr 
                     virus oriP replication origin (Tanaka et al., 1999)"
     primer_bind     complement(6417..6433)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6417..6433)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6430..6452)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6441..6457
                     /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(6465..6471,6472..6489,6490..6495))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6510..6531
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6648..6665)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6819..7407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6899..6918)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7578..8369,8370..8438))
                     /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     8201..8220
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8439..8543)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct gtttaaacaa aaaaataaac gatgcccctt aaagcagaag
       61 ctttaagggg cagagcgttg cggcacatct tttcagacgg ccttattgta gcaacggttc
      121 tcatttcgtt tccgaaatga gaaagggagc tacaaccctt gtggaccaca gtgttaccag
      181 ccggtgtttc gtcctttcca caagatatat aaagccaaga aatcgaaata ctttcaagtt
      241 acggtaagca tatgatagtc cattttaaaa cataatttta aaactgcaaa ctacccaaga
      301 aattattact ttctacgtca cgtattttgt actaatatct ttgtgtttac agtcaaatta
      361 attctaatta tctctctaac agccttgtat cgtatatgca aatatgaagg aatcatggga
      421 aataggccct cttcctgccc gaccttgacg tcgacagatc tcccgatccc ctatggtgca
      481 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
      541 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
      601 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcggaggct
      661 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
      721 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
      781 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
      841 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
      901 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
      961 ccacctggct ccagtacgtg attcttgatc ccgagctgga gccaggggcg ggccttgcgc
     1021 tttaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg
     1081 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat
     1141 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc
     1201 gggccaggat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg
     1261 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
     1321 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
     1381 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
     1441 gccgcttccc ggccctgctc cagggggctc aaaatggagg acgcggcgct cgggagagcg
     1501 ggcgggtgag tcacccacac aaaggaaagg ggcctttccg tcctcagccg tcgcttcatg
     1561 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctgga gcttttggag
     1621 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
     1681 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct
     1741 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
     1801 tccatttcag gtgtcgtgaa caccgccacc atggtgccta agaagaagag aaaggtggct
     1861 gccttcaaac ctaattcaat caactacatc ctcggcctcg ctatcggcat cgcatccgtc
     1921 ggctgggcga tggtagaaat tgacgaagaa gaaaacccca tccgcctgat tgatttgggc
     1981 gtgcgcgtat ttgagcgtgc cgaagtaccg aaaacaggcg actcccttgc catggcaagg
     2041 cgtttggcgc gcagtgttcg ccgcctgacc cgccgtcgcg cccaccgcct gcttcggacc
     2101 cgccgcctat tgaaacgcga aggcgtatta caagccgcca attttgacga aaacggcttg
     2161 attaaatcct taccgaatac accatggcaa cttcgcgcag ccgcattaga ccgcaaactg
     2221 acgcctttag agtggtcggc agtcttgttg catttaatca aacatcgcgg ctatttatcg
     2281 caacggaaaa acgagggcga aactgccgat aaggagcttg gcgctttgct taaaggcgta
     2341 gccggcaatg cccatgcctt acagacaggc gatttccgca caccggccga attggcttta
     2401 aataaatttg agaaagaaag cggccatatc cgcaatcagc gcagcgatta ttcgcatacg
     2461 ttcagccgca aagatttaca ggcggagctg attttgctgt ttgaaaaaca aaaagaattt
     2521 ggcaatccgc atgtttcagg cggccttaaa gaaggtattg aaaccctact gatgacgcaa
     2581 cgccctgccc tgtccggcga tgccgttcaa aaaatgttgg ggcattgcac cttcgaaccg
     2641 gcagagccga aagccgctaa aaacacctac acagccgaac gtttcatctg gctgaccaag
     2701 ctgaacaacc tgcgtatttt agagcaaggc agcgagcggc cattgaccga taccgaacgc
     2761 gccacgctta tggacgagcc atacagaaaa tccaaactga cttacgcaca agcccgtaag
     2821 ctgctgggtt tagaagatac cgcctttttc aaaggcttgc gctatggtaa agacaatgcc
     2881 gaagcctcaa cattgatgga aatgaaggcc taccatgcca tcagccgtgc actggaaaaa
     2941 gaaggattga aagacaaaaa atccccatta aacctttctc ccgaattaca agacgaaatc
     3001 ggcacggcat tctccctgtt caaaaccgat gaagacatta caggccgtct gaaagaccgt
     3061 atacagcccg aaatcttaga agcgctgttg aaacacatca gcttcgataa gttcgtccaa
     3121 atttccttga aagcattgcg ccgaattgtg cctctaatgg aacaaggcaa acgttacgat
     3181 gaagcctgcg ccgaaatcta cggagaccat tacggcaaga agaatacgga agaaaagatt
     3241 tatctgccgc cgattcccgc cgacgaaatc cgcaaccccg tcgtcttgcg cgccttatct
     3301 caagcacgta aggtcattaa cggcgtggta cgccgttacg gctccccagc tcgtatccat
     3361 attgaaactg caagggaagt aggtaaatcg tttaaagacc gcaaagaaat tgagaaacgc
     3421 caagaagaaa accgcaaaga ccgggaaaaa gccgccgcca aattccgaga gtatttcccc
     3481 aattttgtcg gagaacccaa atccaaagat attctgaaac tgcgcctgta cgagcaacaa
     3541 cacggcaaat gcctgtattc gggcaaagaa atcaacttag gccgtctgaa cgaaaaaggc
     3601 tatgtcgaaa tcgaccatgc cctgccgttc tcgcgcacat gggacgacag tttcaacaat
     3661 aaagtactgg tattgggcag cgaaaaccaa aacaaaggca atcaaacccc ttacgaatac
     3721 ttcaacggca aagacaacag ccgcgaatgg caggaattta aagcgcgtgt cgaaaccagc
     3781 cgtttcccgc gcagtaaaaa acaacggatt ctgctgcaaa aattcgatga agacggcttt
     3841 aaagaacgca atctgaacga cacgcgctac gtcaaccgtt tcctgtgtca atttgttgcc
     3901 gaccgtatgc ggctgacagg taaaggcaag aaacgtgtct ttgcatccaa cggacaaatt
     3961 accaatctgt tgcgcggctt ttggggattg cgcaaagtgc gtgcggaaaa cgaccgccat
     4021 cacgccttgg acgccgtcgt cgttgcctgc tcgaccgttg ccatgcagca gaaaattacc
     4081 cgttttgtac gctataaaga gatgaacgcg tttgacggta aaaccataga caaagaaaca
     4141 ggagaagtgc tgcatcaaaa aacacacttc ccacaacctt gggaattttt cgcacaagaa
     4201 gtcatgattc gcgtcttcgg caaaccggac ggcaaacccg aattcgaaga agccgatacc
     4261 ctagaaaaac tgcgcacgtt gcttgccgaa aaattatcat ctcgccccga agccgtacac
     4321 gaatacgtta cgccactgtt tgtttcacgc gcgcccaatc ggaagatgag cgggcaaggg
     4381 catatggaga ccgtcaaatc cgccaaacga ctggacgaag gcgtcagcgt gttgcgcgta
     4441 ccgctgacac agttaaaact gaaagacttg gaaaaaatgg tcaatcggga gcgcgaacct
     4501 aagctatacg aagcactgaa agcacggctg gaagcacata aagacgatcc tgccaaagcc
     4561 tttgccgagc cgttttacaa atacgataaa gcaggcaacc gcacccaaca ggtaaaagcc
     4621 gtacgcgtag agcaagtaca gaaaaccggc gtatgggtgc gcaaccataa cggtattgcc
     4681 gacaacgcaa ccatggtgcg cgtacaccct aatgattaca aataccaaat tgacgaactg
     4741 ggcaaagaaa tcagaccatg ccgtctgaaa aaacgcccgc ctgtccgtta cccatacgat
     4801 gttccagatt acgctgcagc tccagcagcg aagaaaaaga agctggatta actcgctgat
     4861 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     4921 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     4981 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     5041 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     5101 aggcggaaag aaccagctgg ggctcgaccg atgcccttga gagccttcaa cccagtcagc
     5161 tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc
     5221 atgcaactcg taggacaggt gccctggccg gggtcccgca ggaaaaggac aagcagcgaa
     5281 aattcacgcc cccttgggag gtggcggcat atgcaaagga tagcactccc actctactac
     5341 tgggtatcat atgctgactg tatatgcatg aggatagcat atgctacccg gatacagatt
     5401 aggatagcat atactaccca gatatagatt aggatagcat atgctaccca gatatagatt
     5461 aggatagcct atgctaccca gatataaatt aggatagcat atactaccca gatatagatt
     5521 aggatagcat atgctaccca gatatagatt aggatagcct atgctaccca gatatagatt
     5581 aggatagcat atgctaccca gatatagatt aggatagcat atgctatcca gatatttggg
     5641 tagtatatgc tacccagata taaattagga tagcatatac taccctaatc tctattagga
     5701 tagcatatgc tacccggata cagattagga tagcatatac tacccagata tagattagga
     5761 tagcatatgc tacccagata tagattagga tagcctatgc tacccagata taaattagga
     5821 tagcatatac tacccagata tagattagga tagcatatgc tacccagata tagattagga
     5881 tagcctatgc tacccagata tagattagga tagcatatgc tatccagata tttgggtagt
     5941 atatgctatc catggcaaca ttagcccacc gtgctctcag cgacctcgtg aatatgagga
     6001 ccaacaaccc tgtgcttggc gctcaggcgc aagtgtgtgt aatttgtcct ccagatcgca
     6061 gcaatcgcgc ccctatcttg gcccgcccac ctacttatgc aggtattccc cggggtgcca
     6121 ttagtggttt tgtgggcaag tggtttgacc gcagtggtta gcggggttac aatcagccaa
     6181 gttattacac ccttatttta cagtccaaaa ccgcagggcg gcgtgtgggg gctgacgcga
     6241 tcgctgttcc ttaggaccct tttactaacc ctaattcgat agcatatgct tcccgttggg
     6301 taacatatgc tattgaatta gggttagtct ggatagtata tactactacc cgggaagcat
     6361 atgctacccg tttagggttt accgtcgacc tctagctaga gcttggcgta atcatggtca
     6421 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     6481 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     6541 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6601 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6661 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6721 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6781 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6841 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6901 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6961 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7021 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7081 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7141 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7201 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     7261 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7321 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7381 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7441 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7501 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7561 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7621 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7681 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7741 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7801 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7861 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7921 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7981 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8041 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8101 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8161 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8221 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8281 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8341 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8401 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8461 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8521 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga c
//
