LOCUS       sequence-333554-        8269 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of an optimized Cas9 for genome-editing in zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8269)
  AUTHORS   Jao LE, Wente SR, Chen W
  TITLE     Efficient multiplex biallelic zebrafish genome editing using a 
            CRISPR nuclease system.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Aug 5.
  PUBMED    23918387
REFERENCE   2  (bases 1 to 8269)
  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: "Proc Natl 
            Acad Sci U S A. 2013 Aug 5."
COMMENT     Sequence Label: pCS2-nCas9n
FEATURES             Location/Qualifiers
     source          1..8269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..1007
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1042..1060
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1042..1059
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1106..1115
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1124..1144
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1151..5251
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes MGAS15252 Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDDYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFGSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLADSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQIYNQLFEENPINASRVDAKAILSARLSKSRRL
                     ENLIAQLPGEKRNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNSEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGAYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDRGMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGHSLHEQIANLAGSPAIKKGILQTVKIVDELVKVMGH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFIKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             5164..5172
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             5267..5287
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5328..5462
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5352..5371)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5406..5425
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5575..5595)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5575..5593)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5614..5630)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5614..5630)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5627..5649)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5638..5654
                     /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(5662..5668,5669..5686,5687..5692))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5707..5728
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5845..5862)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6016..6604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6096..6115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6775..7566,7567..7635))
                     /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     7398..7417
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7636..7740)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7766..8221)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7903..7924)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8115..8134
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gggcctcttc gctattacgc cagtcgaccg ccaattcaat atggcgtata tggactcatg
       61 ccaattcaat atggtggatc tggacctgtg ccaattcaat atggcgtata tggactcgtg
      121 ccaattcaat atggtggatc tggaccccag ccaattcaat atggcggact tggcaccatg
      181 ccaattcaat atggcggacc tggcactgtg ccaactgggg aggggtctac ttggcacggt
      241 gccaagtttg aggaggggtc ttggccctgt gccaagtccg ccatattgaa ttggcatggt
      301 gccaataatg gcggccatat tggctatatg ccaggatcaa tatataggca atatccaata
      361 tggccctatg ccaatatggc tattggccag gttcaatact atgtattggc cctatgccat
      421 atagtattcc atatatgggt tttcctattg acgtagatag cccctcccaa tgggcggtcc
      481 catataccat atatggggct tcctaatacc gcccatagcc actcccccat tgacgtcaat
      541 ggtctctata tatggtcttt cctattgacg tcatatgggc ggtcctattg acgtatatgg
      601 cgcctccccc attgacgtca attacggtaa atggcccgcc tggctcaatg cccattgacg
      661 tcaataggac cacccaccat tgacgtcaat gggatggctc attgcccatt catatccgtt
      721 ctcacgcccc ctattgacgt caatgacggt aaatggccca cttggcagta catcaatatc
      781 tattaatagt aacttggcaa gtacattact attggaagta cgccagggta cattggcagt
      841 actcccattg acgtcaatgg cggtaaatgg cccgcgatgg ctgccaagta catccccatt
      901 gacgtcaatg gggaggggca atgacgcaaa tgggcgttcc attgacgtaa atgggcggta
      961 ggcgtgccta atgggaggtc tatataagca atgctcgttt agggaaccgc cattctgcct
     1021 ggggacgtcg gagcaagctt gatttaggtg acactataga atacaagcta cttgttcttt
     1081 ttgcaggatc cgatccaatg tcgacgccac catggcttct ccacctaaga agaagagaaa
     1141 ggtgggaagc atggataaga agtatagcat cggcctggat attggaacta actccgtggg
     1201 ttgggcagtg attacagacg actacaaggt ccctagcaag aaatttaagg tgctgggtaa
     1261 caccgacagg cacagcatca agaaaaatct gattggagcc ctgctgttcg gttctggaga
     1321 gactgccgaa gcaacacgcc tgaaaagaac agcaagaagg cgctatacca gaaggaagaa
     1381 tagaatctgt tacctgcagg agattttctc taacgaaatg gctaaggtgg acgattcatt
     1441 ctttcatagg ctggaggaaa gtttcctggt cgaggaagat aagaaacacg agcgccatcc
     1501 tatctttgga aacattgtgg acgaggtcgc ctatcacgaa aaatacccaa ccatctatca
     1561 tctgcgcaag aaactggctg actctactga taaagccgac ctgagactga tctatctggc
     1621 tctggcccac atgattaagt tcaggggtca ttttctgatc gagggcgatc tgaaccccga
     1681 caattccgat gtggacaagc tgttcatcca gctggtccag atttacaatc agctgtttga
     1741 ggaaaaccct attaatgctt ccagagtgga cgcaaaagct atcctgtcag ccaggctgtc
     1801 caagtcacgc agactggaga acctgattgc acagctgccc ggagaaaaga ggaacggtct
     1861 gtttggaaat ctgatcgctc tgagtctggg cctgactcct aacttcaaaa gcaattttga
     1921 tctggctgag gacgccaaac tgcagctgtc aaaggacaca tatgacgatg acctggataa
     1981 cctgctggca cagatcggag atcagtacgc tgacctgttc ctggctgcca aaaatctgtc
     2041 cgacgcaatc ctgctgtcag atattctgag agtgaacagc gagattacaa aagctcctct
     2101 gagtgccagc atgatcaaga gatatgacga gcaccatcag gatctgaccc tgctgaaggc
     2161 tctggtcagg cagcagctgc cagagaagta caaggaaatt ttctttgatc agtccaagaa
     2221 cggctacgcc ggttatatcg acggaggcgc atcacaggag gaattctaca agtttatcaa
     2281 acctattctg gagaagatgg acggaactga ggaactgctg gtgaaactga atagagagga
     2341 cctgctgagg aagcagcgca catttgataa cggttccatc ccacaccaga ttcatctggg
     2401 agagctgcac gctatcctga ggcgccagga agacttctac ccctttctga aagataaccg
     2461 cgagaagatc gaaaaaattc tgaccttcag aatcccttac tatgtgggtc cactggctcg
     2521 cggaaacagc agatttgcct ggatgactcg caaatccgag gaaaccatta ctccttggaa
     2581 cttcgaggaa gtggtcgata agggcgcctc tgcacagtcc ttcatcgaga gaatgactaa
     2641 ttttgacaaa aacctgccca atgagaaagt gctgcctaag cactccctgc tgtacgagta
     2701 tttcactgtc tataacgaac tgacaaaggt gaaatacgtc accgagggca tgagaaagcc
     2761 agccttcctg tcaggagagc agaagaaagc aatcgtggat ctgctgttta aaaccaacag
     2821 gaaagtgact gtcaagcagc tgaaggagga ctacttcaag aaaattgaat gcttcgattc
     2881 cgtggagatc agcggagtcg aagacagatt taacgcaagc ctgggcgctt accacgatct
     2941 gctgaagatc attaaggata aagacttcct ggacaacgag gaaaatgagg atatcctgga
     3001 agacattgtg ctgacactga ccctgtttga ggacagagga atgatcgagg aaagactgaa
     3061 aacctatgct catctgttcg atgacaaggt gatgaaacag ctgaagagaa ggcgctacac
     3121 tggctggggt agactgagca ggaagctgat caacggcatt agggataaac agtcaggaaa
     3181 gacaatcctg gactttctga aaagtgatgg cttcgccaac cgcaatttta tgcagctgat
     3241 tcacgatgac agtctgacct tcaaagagga catccagaag gctcaggtgt ctggacaggg
     3301 ccactccctg catgagcaga ttgcaaacct ggctggaagc ccagccatca agaaaggcat
     3361 tctgcagaca gtgaaaatcg tcgatgagct ggtgaaagtc atgggccata agcccgaaaa
     3421 catcgtgatt gagatggctc gcgaaaatca gacaacccag aagggtcaga agaacagtag
     3481 agagaggatg aaaagaatcg aggaaggcat taaggagctg ggtagccaga tcctgaaaga
     3541 gcacccagtg gaaaacacac agctgcagaa tgagaagctg tatctgtact atctgcagaa
     3601 tggaagagat atgtacgtgg accaggagct ggatattaac aggctgtctg attacgacgt
     3661 ggatcatatc gtcccccaga gtttcatcaa agatgacagc attgacaaca aggtgctgac
     3721 caggtccgac aaaaacagag gaaaatcaga taatgtccct agtgaggaag tggtcaagaa
     3781 aatgaagaac tactggagac agctgctgaa tgccaaactg atcactcaga ggaagtttga
     3841 taacctgaca aaagcagagc gcggtggact gtcagaactg gacaaagctg gattcatcaa
     3901 gaggcagctg gtggaaacac gccagatcac taaacacgtc gcacagattc tggatagtcg
     3961 catgaacaca aagtacgatg agaatgacaa actgatcaga gaagtgaagg tcattaccct
     4021 gaagagtaaa ctggtcagcg actttaggaa agatttccag ttttataagg tccgcgagat
     4081 taacaattat caccatgccc atgacgcata cctgaacgcc gtggtcggta ccgcactgat
     4141 caagaaatac ccaaaactgg agagcgaatt cgtgtacgga gactataagg tgtacgatgt
     4201 cagaaaaatg atcgccaagt ccgagcagga aattggaaaa gctactgcca agtatttctt
     4261 ttactcaaac atcatgaatt tctttaagac agagatcacc ctggccaatg gagaaatccg
     4321 caaaaggccc ctgattgaga caaacggaga gacaggcgaa atcgtgtggg acaaaggcag
     4381 agattttgca accgtgagga aggtcctgag catgcctcaa gtgaatatcg tcaagaaaac
     4441 tgaggtgcag acaggcggtt tctcaaaaga aagtattctg ccaaaacgca actctgataa
     4501 gctgatcgct agaaagaaag actgggaccc taagaagtat ggaggctttg actctcccac
     4561 tgtggcatac tccgtcctgg tggtcgctaa ggtggagaag ggcaaaagca agaaactgaa
     4621 atctgtcaag gagctgctgg gtatcacaat tatggagaga agctctttcg agaagaaccc
     4681 aatcgatttt ctggaggcca aaggttataa ggaagtgaag aaagacctga tcattaaact
     4741 gcccaagtac agtctgtttg agctggaaaa cggcaggaaa cgcatgctgg caagcgctgg
     4801 agagctgcag aaaggcaatg aactggccct gccttctaag tacgtgaact tcctgtatct
     4861 ggcaagccac tacgagaagc tgaaaggatc tccagaggat aacgaacaga aacagctgtt
     4921 tgtggagcag cacaagcatt atctggacga gatcattgaa cagattagcg agttctctaa
     4981 aagagtgatc ctggccgacg caaatctgga taaggtcctg tctgcttaca acaaacacag
     5041 agataagccc atcagggagc aggccgaaaa tatcattcat ctgttcactc tgacaaacct
     5101 gggcgcacct gcagctttca agtacttcga cactacaatc gatagaaaga ggtacacctc
     5161 cactaaggag gtgctggacg ctacactgat ccatcagagt attaccggcc tgtacgaaac
     5221 aaggattgac ctgtctcagc tgggtggcga ctcaccggtg agatctccta agaagaagag
     5281 aaaggtgtga ccgcggaatc tagaactata gtgagtcgta ttacgtagat ccagacatga
     5341 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     5401 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     5461 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     5521 tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta
     5581 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5641 ttatccgctc acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg
     5701 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5761 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5821 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5881 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5941 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6001 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6061 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6121 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6181 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6241 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6301 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6361 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6421 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6481 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6541 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6601 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6661 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6721 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6781 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6841 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6901 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6961 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7021 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7081 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7141 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7201 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7261 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7321 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7381 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7441 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7501 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7561 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7621 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7681 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7741 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     7801 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     7861 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     7921 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     7981 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     8041 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     8101 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     8161 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     8221 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgc
//
