LOCUS       sequence-221986-        8877 bp    DNA     circular SYN 15-DEC-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 8877)
  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 8877)
  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: "Genome 
            Biol."; date: "2018-12-5"; volume: "19(1):214. doi"; pages: " 
            10.1186/s13059-018-1591-1"
COMMENT     Sequence Label: pSimpleII-U6-sgRNA-BsmBI-NLS-NmCas9-HA-NLS
FEATURES             Location/Qualifiers
     source          1..8877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        complement(36..128)
                     /label=Nm tracrRNA
                     /note="trans-activating CRISPR RNA for the Neisseria 
                     meningitidis CRISPR/Cas9 system (Hou et al., 2013)"
     promoter        complement(177..425)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(235..254)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(405..425)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(482..501)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        651..1813
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          882..1804
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1761..1781
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1819..1828
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1831..1851
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1852..5094
                     /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="AAFKPNSINYILGLDIGIASVGWAMVEIDEEENPIRLIDLGVRVF
                     ERAEVPKTGDSLAMARRLARSVRRLTRRRAHRLLRTRRLLKREGVLQAANFDENGLIKS
                     LPNTPWQLRAAALDRKLTPLEWSAVLLHLIKHRGYLSQRKNEGETADKELGALLKGVAG
                     NAHALQTGDFRTPAELALNKFEKESGHIRNQRSDYSHTFSRKDLQAELILLFEKQKEFG
                     NPHVSGGLKEGIETLLMTQRPALSGDAVQKMLGHCTFEPAEPKAAKNTYTAERFIWLTK
                     LNNLRILEQGSERPLTDTERATLMDEPYRKSKLTYAQARKLLGLEDTAFFKGLRYGKDN
                     AEASTLMEMKAYHAISRALEKEGLKDKKSPLNLSPELQDEIGTAFSLFKTDEDITGRLK
                     DRIQPEILEALLKHISFDKFVQISLKALRRIVPLMEQGKRYDEACAEIYGDHYGKKNTE
                     EKIYLPPIPADEIRNPVVLRALSQARKVINGVVRRYGSPARIHIETAREVGKSFKDRKE
                     IEKRQEENRKDREKAAAKFREYFPNFVGEPKSKDILKLRLYEQQHGKCLYSGKEINLGR
                     LNEKGYVEIDHALPFSRTWDDSFNNKVLVLGSENQNKGNQTPYEYFNGKDNSREWQEFK
                     ARVETSRFPRSKKQRILLQKFDEDGFKERNLNDTRYVNRFLCQFVADRMRLTGKGKKRV
                     FASNGQITNLLRGFWGLRKVRAENDRHHALDAVVVACSTVAMQQKITRFVRYKEMNAFD
                     GKTIDKETGEVLHQKTHFPQPWEFFAQEVMIRVFGKPDGKPEFEEADTLEKLRTLLAEK
                     LSSRPEAVHEYVTPLFVSRAPNRKMSGQGHMETVKSAKRLDEGVSVLRVPLTQLKLKDL
                     EKMVNREREPKLYEALKARLEAHKDDPAKAFAEPFYKYDKAGNRTQQVKAVRVEQVQKT
                     GVWVRNHNGIADNATMVRVDVFEKGDKYYLVPIYSWQVAKGILPDRAVVQGKDEEDWQL
                     IDDSFNFKFSLHPNDLVEVITKKARMFGYFASCHRGTGNINIRIHDLDHKIGKNGILEG
                     IGVKTALSFQKYQIDELGKEIRPCRLKKRPPVR"
     CDS             5095..5121
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5128..5154
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     primer_bind     complement(5170..5187)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5176..5400
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5565..6678
                     /label=mini-oriP
                     /note="internally truncated variant of the Epstein-Barr 
                     virus oriP replication origin (Tanaka et al., 1999)"
     primer_bind     complement(6723..6739)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6723..6739)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6736..6758)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6747..6763
                     /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(6771..6777,6778..6795,6796..6801))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6816..6837
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6954..6971)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7125..7713)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7205..7224)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7884..8675,8676..8744))
                     /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     8507..8526
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8745..8849)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct gtttaaacaa aaaaataaac gatgcccctt aaagcagaag
       61 ctttaagggg cagagcgttg cggcacatct tttcagacgg ccttattgta gcaacggttc
      121 tcatttcgtt tccgaaatga gaaagggagc tacaactgag acgtggaatc gtctccggtg
      181 tttcgtcctt tccacaagat atataaagcc aagaaatcga aatactttca agttacggta
      241 agcatatgat agtccatttt aaaacataat tttaaaactg caaactaccc aagaaattat
      301 tactttctac gtcacgtatt ttgtactaat atctttgtgt ttacagtcaa attaattcta
      361 attatctctc taacagcctt gtatcgtata tgcaaatatg aaggaatcat gggaaatagg
      421 ccctcttcct gcccgacctt gacgtcgaca gatctcccga tcccctatgg tgcactctca
      481 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
      541 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
      601 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgga ggctccggtg
      661 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      721 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      781 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      841 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      901 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      961 ggctccagta cgtgattctt gatcccgagc tggagccagg ggcgggcctt gcgctttagg
     1021 agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg
     1081 aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa
     1141 tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca
     1201 ggatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc
     1261 ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg
     1321 gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc
     1381 gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct
     1441 tcccggccct gctccagggg gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg
     1501 tgagtcaccc acacaaagga aaggggcctt tccgtcctca gccgtcgctt catgtgactc
     1561 cacggagtac cgggcgccgt ccaggcacct cgattagttc tggagctttt ggagtacgtc
     1621 gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga
     1681 gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga
     1741 gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt
     1801 tcaggtgtcg tgaacaccgc caccatggtg cctaagaaga agagaaaggt ggctgccttc
     1861 aaacctaatt caatcaacta catcctcggc ctcgatatcg gcatcgcatc cgtcggctgg
     1921 gcgatggtag aaattgacga agaagaaaac cccatccgcc tgattgattt gggcgtgcgc
     1981 gtatttgagc gtgccgaagt accgaaaaca ggcgactccc ttgccatggc aaggcgtttg
     2041 gcgcgcagtg ttcgccgcct gacccgccgt cgcgcccacc gcctgcttcg gacccgccgc
     2101 ctattgaaac gcgaaggcgt attacaagcc gccaattttg acgaaaacgg cttgattaaa
     2161 tccttaccga atacaccatg gcaacttcgc gcagccgcat tagaccgcaa actgacgcct
     2221 ttagagtggt cggcagtctt gttgcattta atcaaacatc gcggctattt atcgcaacgg
     2281 aaaaacgagg gcgaaactgc cgataaggag cttggcgctt tgcttaaagg cgtagccggc
     2341 aatgcccatg ccttacagac aggcgatttc cgcacaccgg ccgaattggc tttaaataaa
     2401 tttgagaaag aaagcggcca tatccgcaat cagcgcagcg attattcgca tacgttcagc
     2461 cgcaaagatt tacaggcgga gctgattttg ctgtttgaaa aacaaaaaga atttggcaat
     2521 ccgcatgttt caggcggcct taaagaaggt attgaaaccc tactgatgac gcaacgccct
     2581 gccctgtccg gcgatgccgt tcaaaaaatg ttggggcatt gcaccttcga accggcagag
     2641 ccgaaagccg ctaaaaacac ctacacagcc gaacgtttca tctggctgac caagctgaac
     2701 aacctgcgta ttttagagca aggcagcgag cggccattga ccgataccga acgcgccacg
     2761 cttatggacg agccatacag aaaatccaaa ctgacttacg cacaagcccg taagctgctg
     2821 ggtttagaag ataccgcctt tttcaaaggc ttgcgctatg gtaaagacaa tgccgaagcc
     2881 tcaacattga tggaaatgaa ggcctaccat gccatcagcc gtgcactgga aaaagaagga
     2941 ttgaaagaca aaaaatcccc attaaacctt tctcccgaat tacaagacga aatcggcacg
     3001 gcattctccc tgttcaaaac cgatgaagac attacaggcc gtctgaaaga ccgtatacag
     3061 cccgaaatct tagaagcgct gttgaaacac atcagcttcg ataagttcgt ccaaatttcc
     3121 ttgaaagcat tgcgccgaat tgtgcctcta atggaacaag gcaaacgtta cgatgaagcc
     3181 tgcgccgaaa tctacggaga ccattacggc aagaagaata cggaagaaaa gatttatctg
     3241 ccgccgattc ccgccgacga aatccgcaac cccgtcgtct tgcgcgcctt atctcaagca
     3301 cgtaaggtca ttaacggcgt ggtacgccgt tacggctccc cagctcgtat ccatattgaa
     3361 actgcaaggg aagtaggtaa atcgtttaaa gaccgcaaag aaattgagaa acgccaagaa
     3421 gaaaaccgca aagaccggga aaaagccgcc gccaaattcc gagagtattt ccccaatttt
     3481 gtcggagaac ccaaatccaa agatattctg aaactgcgcc tgtacgagca acaacacggc
     3541 aaatgcctgt attcgggcaa agaaatcaac ttaggccgtc tgaacgaaaa aggctatgtc
     3601 gaaatcgacc atgccctgcc gttctcgcgc acatgggacg acagtttcaa caataaagta
     3661 ctggtattgg gcagcgaaaa ccaaaacaaa ggcaatcaaa ccccttacga atacttcaac
     3721 ggcaaagaca acagccgcga atggcaggaa tttaaagcgc gtgtcgaaac cagccgtttc
     3781 ccgcgcagta aaaaacaacg gattctgctg caaaaattcg atgaagacgg ctttaaagaa
     3841 cgcaatctga acgacacgcg ctacgtcaac cgtttcctgt gtcaatttgt tgccgaccgt
     3901 atgcggctga caggtaaagg caagaaacgt gtctttgcat ccaacggaca aattaccaat
     3961 ctgttgcgcg gcttttgggg attgcgcaaa gtgcgtgcgg aaaacgaccg ccatcacgcc
     4021 ttggacgccg tcgtcgttgc ctgctcgacc gttgccatgc agcagaaaat tacccgtttt
     4081 gtacgctata aagagatgaa cgcgtttgac ggtaaaacca tagacaaaga aacaggagaa
     4141 gtgctgcatc aaaaaacaca cttcccacaa ccttgggaat ttttcgcaca agaagtcatg
     4201 attcgcgtct tcggcaaacc ggacggcaaa cccgaattcg aagaagccga taccctagaa
     4261 aaactgcgca cgttgcttgc cgaaaaatta tcatctcgcc ccgaagccgt acacgaatac
     4321 gttacgccac tgtttgtttc acgcgcgccc aatcggaaga tgagcgggca agggcatatg
     4381 gagaccgtca aatccgccaa acgactggac gaaggcgtca gcgtgttgcg cgtaccgctg
     4441 acacagttaa aactgaaaga cttggaaaaa atggtcaatc gggagcgcga acctaagcta
     4501 tacgaagcac tgaaagcacg gctggaagca cataaagacg atcctgccaa agcctttgcc
     4561 gagccgtttt acaaatacga taaagcaggc aaccgcaccc aacaggtaaa agccgtacgc
     4621 gtagagcaag tacagaaaac cggcgtatgg gtgcgcaacc ataacggtat tgccgacaac
     4681 gcaaccatgg tgcgcgtaga tgtgtttgag aaaggcgaca agtattatct ggtaccgatt
     4741 tacagttggc aggtagcgaa agggattttg ccggataggg ctgttgtaca aggaaaagat
     4801 gaagaagatt ggcaacttat tgatgatagt ttcaacttta aattctcatt acaccctaat
     4861 gatttagtcg aggttataac aaaaaaagct agaatgtttg gttactttgc cagctgccat
     4921 cgaggcacag gtaatatcaa tatacgcatt catgatcttg atcataaaat tggcaaaaat
     4981 ggaatactgg aaggtatcgg cgtcaaaacc gccctttcat tccaaaaata ccaaattgac
     5041 gaactgggca aagaaatcag accatgccgt ctgaaaaaac gcccgcctgt ccgttaccca
     5101 tacgatgttc cagattacgc tgcagctcca gcagcgaaga aaaagaagct ggattaactc
     5161 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     5221 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     5281 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     5341 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     5401 cttctgaggc ggaaagaacc agctggggct cgaccgatgc ccttgagagc cttcaaccca
     5461 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
     5521 tttatcatgc aactcgtagg acaggtgccc tggccggggt cccgcaggaa aaggacaagc
     5581 agcgaaaatt cacgccccct tgggaggtgg cggcatatgc aaaggatagc actcccactc
     5641 tactactggg tatcatatgc tgactgtata tgcatgagga tagcatatgc tacccggata
     5701 cagattagga tagcatatac tacccagata tagattagga tagcatatgc tacccagata
     5761 tagattagga tagcctatgc tacccagata taaattagga tagcatatac tacccagata
     5821 tagattagga tagcatatgc tacccagata tagattagga tagcctatgc tacccagata
     5881 tagattagga tagcatatgc tacccagata tagattagga tagcatatgc tatccagata
     5941 tttgggtagt atatgctacc cagatataaa ttaggatagc atatactacc ctaatctcta
     6001 ttaggatagc atatgctacc cggatacaga ttaggatagc atatactacc cagatataga
     6061 ttaggatagc atatgctacc cagatataga ttaggatagc ctatgctacc cagatataaa
     6121 ttaggatagc atatactacc cagatataga ttaggatagc atatgctacc cagatataga
     6181 ttaggatagc ctatgctacc cagatataga ttaggatagc atatgctatc cagatatttg
     6241 ggtagtatat gctatccatg gcaacattag cccaccgtgc tctcagcgac ctcgtgaata
     6301 tgaggaccaa caaccctgtg cttggcgctc aggcgcaagt gtgtgtaatt tgtcctccag
     6361 atcgcagcaa tcgcgcccct atcttggccc gcccacctac ttatgcaggt attccccggg
     6421 gtgccattag tggttttgtg ggcaagtggt ttgaccgcag tggttagcgg ggttacaatc
     6481 agccaagtta ttacaccctt attttacagt ccaaaaccgc agggcggcgt gtgggggctg
     6541 acgcgatcgc tgttccttag gaccctttta ctaaccctaa ttcgatagca tatgcttccc
     6601 gttgggtaac atatgctatt gaattagggt tagtctggat agtatatact actacccggg
     6661 aagcatatgc tacccgttta gggtttaccg tcgacctcta gctagagctt ggcgtaatca
     6721 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6781 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6841 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6901 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6961 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     7021 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     7081 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     7141 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     7201 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     7261 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     7321 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     7381 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     7441 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7501 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7561 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7621 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7681 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7741 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7801 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7861 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7921 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7981 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     8041 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     8101 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     8161 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     8221 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     8281 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     8341 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     8401 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     8461 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     8521 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     8581 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8641 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8701 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8761 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8821 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgac
//
