pSaCas9_GFP

LOCUS       pSaCas9_GFP                8398 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Co-expression of human codon-optimized Staphylococcus aureus Cas9 
            nuclease and GFP, plasmid optimized for expression in human 
            pluripotent stem cells and other mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pSaCas9_GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8398)
  TITLE     Staphylococcus aureus Cas9
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8398)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8398
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        8..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..665
                     /label=chicken beta-actin promoter
     intron          666..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1739..1748
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1748..1813
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1820..1840
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1850..5008
                     /codon_start=1
                     /product="Cas9 endonuclease from the Staphylococcus aureus 
                     Type II CRISPR/Cas system"
                     /label=SaCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MKRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNE
                     GRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEE
                     FSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGE
                     VRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFG
                     WKDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKF
                     QIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKE
                     IIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKA
                     INLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSI
                     KVINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENA
                     KYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVK
                     QEENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRF
                     SVQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKER
                     NKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEI
                     FITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKD
                     NDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKK
                     DNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKN
                     LDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLL
                     NRIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQ
                     IIKKG"
     CDS             5024..5077
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5078..5797
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5123..5144)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5384..5403)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5731..5752
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5840..5961
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5877..5896)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5931..5950
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6010..6026)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6010..6026)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6023..6045)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6034..6050
                     /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(6058..6088)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6103..6124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6241..6258)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6412..7000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6492..6511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7171..8031)
                     /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     7794..7813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8032..8136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8215..8234)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gactatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatgggt cgaggtgagc cccacgttct gcttcactct
      421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      481 tgcagcgatg ggggcggggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg
      541 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
      601 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
      661 gggcgggagt cgctgcgttg ccttcgcccc gtgccccgct ccgcgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggctcgt ttcttttctg tggctgcgtg
      841 aaagccttaa agggctccgg gagggccctt tgtgcggggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggcccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcgtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccat ctccagcctc ggggctgtcc
     1561 gcagggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagtgcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcgcggccgc
     1741 caccatggac tataaggacc acgacggaga ctacaaggat catgatattg attacaaaga
     1801 cgatgacgat aagatggccc cgaagaaaaa gcgcaaggtc gaagcgtcca tgaaaaggaa
     1861 ctacattctg gggctggaca tcgggattac aagcgtgggg tatgggatta ttgactatga
     1921 aacaagggac gtgatcgacg caggcgtcag actgttcaag gaggccaacg tggaaaacaa
     1981 tgagggacgg agaagcaaga ggggagccag gcgcctgaaa cgacggagaa ggcacagaat
     2041 ccagagggtg aagaaactgc tgttcgatta caacctgctg accgaccatt ctgagctgag
     2101 tggaattaat ccttatgaag ccagggtgaa aggcctgagt cagaagctgt cagaggaaga
     2161 gttttccgca gctctgctgc acctggctaa gcgccgagga gtgcataacg tcaatgaggt
     2221 ggaagaggac accggcaacg agctgtctac aaaggaacag atctcacgca atagcaaagc
     2281 tctggaagag aagtatgtcg cagagctaca gctggaacgg ctgaagaaag atggcgaggt
     2341 gagagggtca attaataggt tcaagacaag cgactacgtc aaagaagcca agcagctgct
     2401 gaaagtgcag aaggcttacc accagctgga tcagagcttc atcgatactt atatcgacct
     2461 gctggagact cggagaacct actatgaggg accaggagaa gggagcccct tcggatggaa
     2521 agacatcaag gaatggtacg agatgctgat gggacattgc acctattttc cagaagagct
     2581 gagaagcgtc aagtacgctt ataacgcaga tctgtacaac gccctgaatg acctgaacaa
     2641 cctggtcatc accagggatg aaaacgagaa actggaatac tatgagaagt tccagatcat
     2701 cgaaaacgtg tttaagcaga agaaaaagcc tacactgaaa cagattgcta aggagatcct
     2761 ggtcaacgaa gaggacatca agggctaccg ggtgacaagc actggaaaac cagagttcac
     2821 caatctgaaa gtgtatcacg atattaagga catcacagca cggaaagaaa tcattgagaa
     2881 cgccgaactg ctggatcaga ttgctaagat cctgactatc taccagagtt ccgaggacat
     2941 ccaggaagag ctgactaacc tgaacagcga gctgacccag gaagagatcg aacagattag
     3001 taatctgaag gggtacaccg gaacacacaa cctgtccctg aaagctatca atctgattct
     3061 ggatgagctg tggcatacaa acgacaatca gattgcaatc tttaaccggc tgaagctggt
     3121 accaaaaaag gtggacctga gtcagcagaa agagatccca accacactgg tggacgattt
     3181 cattctgtca cccgtggtca agcggagctt catccagagc atcaaagtga tcaacgccat
     3241 catcaagaag tacggcctgc ccaatgatat cattatcgag ctggctaggg agaagaacag
     3301 caaggacgca cagaagatga tcaatgagat gcagaaacga aaccggcaga ccaatgaacg
     3361 cattgaagag attatccgaa ctaccgggaa agagaacgca aagtacctga ttgaaaaaat
     3421 caagctgcac gatatgcagg agggaaagtg tctgtattct ctggaggcca tccccctgga
     3481 ggacctgctg aacaatccat tcaactacga ggtcgatcat attatcccca gaagcgtgtc
     3541 cttcgacaat tcctttaaca acaaggtgct ggtcaagcag gaagagaact ctaaaaaggg
     3601 caataggact cctttccagt acctgtctag ttcagattcc aagatctctt acgaaacctt
     3661 taaaaagcac attctgaatc tggccaaagg aaagggccgc atcagcaaga ccaaaaagga
     3721 gtacctgctg gaagagcggg acatcaacag attctccgtc cagaaggatt ttattaaccg
     3781 gaatctggtg gacacaagat acgctactcg cggcctgatg aatctgctgc gatcctattt
     3841 ccgggtgaac aatctggatg tgaaagtcaa gtccatcaac ggcgggttca catcttttct
     3901 gaggcgcaaa tggaagttta aaaaggagcg caacaaaggg tacaagcacc atgccgaaga
     3961 tgctctgatt atcgcaaatg ccgacttcat ctttaaggag tggaaaaagc tggacaaagc
     4021 caagaaagtg atggagaacc agatgttcga agagaagcag gccgaatcta tgcccgaaat
     4081 cgagacagaa caggagtaca aggagatttt catcactcct caccagatca agcatatcaa
     4141 ggatttcaag gactacaagt actctcaccg ggtggataaa aagcccaaca gagagctgat
     4201 caatgacacc ctgtatagta caagaaaaga cgataagggg aataccctga ttgtgaacaa
     4261 tctgaacgga ctgtacgaca aagataatga caagctgaaa aagctgatca acaaaagtcc
     4321 cgagaagctg ctgatgtacc accatgatcc tcagacatat cagaaactga agctgattat
     4381 ggagcagtac ggcgacgaga agaacccact gtataagtac tatgaagaga ctgggaacta
     4441 cctgaccaag tatagcaaaa aggataatgg ccccgtgatc aagaagatca agtactatgg
     4501 gaacaagctg aatgcccatc tggacatcac agacgattac cctaacagtc gcaacaaggt
     4561 ggtcaagctg tcactgaagc catacagatt cgatgtctat ctggacaacg gcgtgtataa
     4621 atttgtgact gtcaagaatc tggatgtcat caaaaaggag aactactatg aagtgaatag
     4681 caagtgctac gaagaggcta aaaagctgaa aaagattagc aaccaggcag agttcatcgc
     4741 ctccttttac aacaacgacc tgattaagat caatggcgaa ctgtataggg tcatcggggt
     4801 gaacaatgat ctgctgaacc gcattgaagt gaatatgatt gacatcactt accgagagta
     4861 tctggaaaac atgaatgata agcgcccccc tcgaattatc aaaacaatcg cctctaagac
     4921 tcagagtatc aaaaagtact caaccgacat tctgggaaac ctgtatgagg tgaagagcaa
     4981 aaagcaccct cagattatca aaaagggcag gtccggcggc ggagagggca gaggaagtct
     5041 tctaacatgc ggtgacgtgg aggagaatcc cggcccaatg gtgagcaagg gcgaggagct
     5101 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     5161 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     5221 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     5281 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     5341 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     5401 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     5461 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     5521 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     5581 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     5641 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     5701 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     5761 cgggatcact ctcggcatgg acgagctgta caagtaactg cagcgcgggg atctcatgct
     5821 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5881 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5941 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     6001 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6061 catacgagcc ggaagcataa agtgtaaagc ctagggtgcc taatgagtga gctaactcac
     6121 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6181 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6241 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6301 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6361 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6421 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6481 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6541 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6601 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6661 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6721 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6781 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6841 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6901 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6961 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7021 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7081 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7141 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7201 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7261 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7321 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7381 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7441 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7501 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7561 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7621 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7681 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7741 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7801 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7861 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7921 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7981 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8041 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8101 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8161 tgacgtcgac ggatcgggag atcgatctcc cgatccccta gggtcgactc tcagtacaat
     8221 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     8281 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     8341 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatat
//