iSpyMac

LOCUS       iSpyMac               10111 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses the codon-optimized, engineered iSpyMac Cas9 nuclease in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    iSpyMac
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10111)
  AUTHORS   Chatterjee P, Lee J, Nip L, Koseki SRT, Tysinger E, Sontheimer EJ, 
            Jacobson JM, Jakimo N
  TITLE     A Cas9 with PAM recognition for adenine dinucleotides.
  JOURNAL   Nat Commun. 2020 May 18;11(1):2474. doi: 10.1038/s41467-020-16117-8.
  PUBMED    32424114
REFERENCE   2  (bases 1 to 10111)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..10111
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1292..3010
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRKL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLKREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     CDS             5426..5446
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5456..5512
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5513..6229
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5558..5579)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5819..5838)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6166..6187
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6242..6262
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6271..6288
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    6321..6909
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6374..6394)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6792..6803)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6981..7214
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7292..7413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7329..7348)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7383..7402
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7453..7588
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7515..7534
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(7608..7627)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7609..7627)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7637..7654)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7637..7653)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7760..7777)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7931..8519)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8011..8030)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8690..9550)
                     /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     9313..9332
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9551..9655)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9734..9753)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_feature    9887..9958
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      9963..10009
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    10027..10075
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 ctagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcgt ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgtggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat cagcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg cccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagcac aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcca gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgaa aactacccct aaaagccaaa
     1201 agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgct gccaagcttc
     1261 cgagctctcg aattcaaagg aggtacccac catggacaag aagtacagca tcggcctgga
     1321 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
     1381 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
     1441 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
     1501 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
     1561 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
     1621 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
     1681 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
     1741 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
     1801 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
     1861 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
     1921 catcctgtct gccagactga gcaagagcag aaagctggaa aatctgatcg cccagctgcc
     1981 cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc
     2041 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
     2101 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
     2161 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
     2221 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
     2281 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
     2341 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
     2401 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
     2461 cgtgaagctg aagagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
     2521 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
     2581 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
     2641 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
     2701 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
     2761 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
     2821 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
     2881 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
     2941 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
     3001 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
     3061 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
     3121 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
     3181 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
     3241 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
     3301 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
     3361 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
     3421 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
     3481 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
     3541 gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
     3601 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
     3661 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
     3721 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
     3781 caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga
     3841 ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt
     3901 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
     3961 gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
     4021 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
     4081 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
     4141 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
     4201 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctaaa
     4261 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta
     4321 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg
     4381 caaggctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
     4441 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
     4501 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
     4561 ccaagtgaat atcgtgaaaa agaccgagat ccagaccgtg ggacagaatg gaggcctgtt
     4621 cgacgacaac cccaagagcc ccctggaggt gacccccagc aaactggtgc ccctgaagaa
     4681 agagctgaac cccaagaaat acggcggcta ccagaagccc accaccgcct accccgtgct
     4741 gctgatcacc gacaccaagc agctgatccc catcagcgtg atgaacaaga aacagttcga
     4801 gcagaacccc gtgaagttcc tgcgggaccg gggctaccag caggtgggca agaacgactt
     4861 catcaagctg cccaagtaca ccctggtgga catcggcgac ggcatcaagc ggctgtgggc
     4921 cagcagcaag gagatccaca agggcaacca gctggtggtg agcaagaaga gccagatcct
     4981 gctgtaccac gcccaccacc tggacagcga cctgagcaac gactacctgc agaaccacaa
     5041 ccagcagttc gacgtgctgt tcaacgagat catcagcttc agcaaaaagt gcaagctggg
     5101 caaggagcac atccagaaga tcgagaacgt gtacagcaac aagaagaaca gcgccagcat
     5161 cgaggagctg gccgagagct ttatcaaact gctcggcttc acccagctgg gcgccaccag
     5221 ccccttcaac ttcctgggcg tgaagctgaa ccagaagcag tacaagggca agaaggacta
     5281 catcctgccc tgcaccgagg gcaccctgat cagacagagc atcaccggac tgtacgagac
     5341 cagagtggat ctgagcaaga tcggcgagga tagcggcgga agcggaggca gcaagagaac
     5401 cgccgatgga agcgagttcg agtctcccaa gaagaagagg aaagtcggaa gcggagctac
     5461 taacttcagc ctgctgaagc aggctggaga cgtggaggag aaccctggac ctatggtgag
     5521 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     5581 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     5641 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     5701 caccctgacc tatggagtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     5761 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     5821 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     5881 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     5941 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     6001 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     6061 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     6121 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     6181 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt ctggtggttc
     6241 tcccaagaag aaaaggaaag tctaaccggt catcatcacc atcaccattg agtttaaacc
     6301 cgctgatcag caaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     6361 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     6421 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     6481 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     6541 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     6601 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     6661 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     6721 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     6781 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     6841 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     6901 cctccccgcg tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     6961 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct
     7021 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     7081 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     7141 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     7201 gaaaatctct agcagtacgt atagtagttc atgtcatctt attattcagt atttataact
     7261 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     7321 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     7381 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     7441 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     7501 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     7561 aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt
     7621 cgtattacgc gcgctcactg gccgtcgttt tacactcact gcccgctttc cagtcgggaa
     7681 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc gggaagaggc ggtttgcgta
     7741 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     7801 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     7861 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     7921 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     7981 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     8041 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     8101 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     8161 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     8221 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     8281 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     8341 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     8401 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     8461 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     8521 aagatccttt gatcttttct acggggtctg acactcagtg gaacgaaaac tcacgttaag
     8581 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     8641 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     8701 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     8761 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     8821 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     8881 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     8941 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     9001 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     9061 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     9121 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     9181 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     9241 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     9301 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     9361 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     9421 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     9481 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     9541 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     9601 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     9661 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tcgatctccc gatcccctag
     9721 ggtcgactct cagtacaatc tgctctgatg ccgctcgact ctggctctat cgaatctccg
     9781 tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg
     9841 tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt
     9901 agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa
     9961 ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc
    10021 gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta
    10081 actgctgatc gagtgtagcc agatctcgtg a
//