LOCUS       sequence-330622-        8724 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of a zebrafish optimized Cas9 driven by a tilapia 
            promoter in fish cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8724)
  AUTHORS   Hamar J, Kultz D
  TITLE     An efficient vector-based CRISPR/Cas9 system in an Oreochromis 
            mossambicus cell line using endogenous promoters.
  JOURNAL   Sci Rep. 2021 Apr 12;11(1):7854. doi: 10.1038/s41598-021-87068-3.
  PUBMED    33846462
REFERENCE   2  (bases 1 to 8724)
  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: "Sci Rep."; 
            date: "2021-04-12"; volume: "11(1):7854. doi"; pages: " 
            10.1038/s41598-021-87068-3"
COMMENT     Sequence Label: OmEF1aCas9P2ApuroSB
FEATURES             Location/Qualifiers
     source          1..8724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1575..1595
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1602..5702
                     /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             5615..5623
                     /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             5733..5789
                     /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             5790..6389
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5790..5809)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6286..6306
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6489..6508)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6554..6609
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6934..6951)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7105..7693)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7185..7204)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7864..8655,8656..8724))
                     /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     8487..8506
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct gatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
      181 ggaaattgta agcgttaata ttttgttaaa attggatccc tatacagttg aagtcggaag
      241 tttacataca cttaagttgg agtcattaaa actcgttttt caactactcc acaaatttct
      301 tgttaacaaa caatagtttt ggcaagtcag ttaggacatc tactttgtgc atgacacaag
      361 tcatttttcc aacaattgtt tacagacaga ttatttcact tataattcac tgtatcacaa
      421 ttccagtggg tcagaagttt acatacacta agttcgactc ctctgcagaa tgcggcgatg
      481 tttcggtaag gggtccgcta tctagtaggc ccagtcgagg tcgacgttgc agggtttcat
      541 tcggcggcgt gctgacgttt tttgcaggaa tgtgtgtgat ttgcggggtt ttgggccggg
      601 gagggtgacg ggaaatgagg ccgcctccca ttgagtataa aacctcccag tccggcccct
      661 gtcttcctct ttctgttacc tggcttaggg gcagcagctt tggccgtgaa aacccagaaa
      721 caccggtgag tttagctccc attcttcaca ttgagaggga cccggcctcg ggcctttaaa
      781 ggttttagtt aaacgtgatt tgttcaggcg ttagcatcct ctttgtgctt ctttctttta
      841 ctcccaaact ttgctttttc ggttacgtaa cttgacacta acctgtagat ttaattttcg
      901 agggaattaa ccctaaaaaa aatagtaaag gtcgttctcg cctgattgta tttaacatga
      961 tagttttttg tgaggtttgt gcgttagtgg atttactaga tgctgttttt taaacttttg
     1021 acgaaagatt ttttacgagg gaggagaaaa tgtagagtgg gcaacgggaa caaaggaggg
     1081 ctgcaagacc ggagatccga agatgcttga atggcatttc ttctctctcc tcactgaaac
     1141 aactgcccgt atttatccgg caacaggtgc ttgaagtccg gacaagatgg ctttgaatat
     1201 ttggcgatct ttgtaacttt atccgacgtg gcccagtgga gtaacacgtg tctcggctgg
     1261 cttgtggccg agtgaggcct agcagtagca gccgcctggt agcggctaaa gcctagcgac
     1321 tgggtagcca ttgagttttt aacttagtat aaattaaaaa aaaaacaata cgttttagta
     1381 ggctttcttt ttttttttta aacgcctttc attaggccta tatgccggga tggtttccta
     1441 atgtagctgg gctcaagatg cgatggttta gttgtacatg cacatctgtt aacgaagctt
     1501 gacggctact taatattaag cctaacgcaa tttttattta cttgcccaca gaaactaaaa
     1561 ccatggcttc tccacctaag aagaagagaa aggtgggaag catggataag aagtatagca
     1621 tcggcctgga tattggaact aactccgtgg gttgggcagt gattacagac gactacaagg
     1681 tccctagcaa gaaatttaag gtgctgggta acaccgacag gcacagcatc aagaaaaatc
     1741 tgattggagc cctgctgttc ggttctggag agactgccga agcaacacgc ctgaaaagaa
     1801 cagcaagaag gcgctatacc agaaggaaga atagaatctg ttacctgcag gagattttct
     1861 ctaacgaaat ggctaaggtg gacgattcat tctttcatag gctggaggaa agtttcctgg
     1921 tcgaggaaga taagaaacac gagcgccatc ctatctttgg aaacattgtg gacgaggtcg
     1981 cctatcacga aaaataccca accatctatc atctgcgcaa gaaactggct gactctactg
     2041 ataaagccga cctgagactg atctatctgg ctctggccca catgattaag ttcaggggtc
     2101 attttctgat cgagggcgat ctgaaccccg acaattccga tgtggacaag ctgttcatcc
     2161 agctggtcca gatttacaat cagctgtttg aggaaaaccc tattaatgct tccagagtgg
     2221 acgcaaaagc tatcctgtca gccaggctgt ccaagtcacg cagactggag aacctgattg
     2281 cacagctgcc cggagaaaag aggaacggtc tgtttggaaa tctgatcgct ctgagtctgg
     2341 gcctgactcc taacttcaaa agcaattttg atctggctga ggacgccaaa ctgcagctgt
     2401 caaaggacac atatgacgat gacctggata acctgctggc acagatcgga gatcagtacg
     2461 ctgacctgtt cctggctgcc aaaaatctgt ccgacgcaat cctgctgtca gatattctga
     2521 gagtgaacag cgagattaca aaagctcctc tgagtgccag catgatcaag agatatgacg
     2581 agcaccatca ggatctgacc ctgctgaagg ctctggtcag gcagcagctg ccagagaagt
     2641 acaaggaaat tttctttgat cagtccaaga acggctacgc cggttatatc gacggaggcg
     2701 catcacagga ggaattctac aagtttatca aacctattct ggagaagatg gacggaactg
     2761 aggaactgct ggtgaaactg aatagagagg acctgctgag gaagcagcgc acatttgata
     2821 acggttccat cccacaccag attcatctgg gagagctgca cgctatcctg aggcgccagg
     2881 aagacttcta cccctttctg aaagataacc gcgagaagat cgaaaaaatt ctgaccttca
     2941 gaatccctta ctatgtgggt ccactggctc gcggaaacag cagatttgcc tggatgactc
     3001 gcaaatccga ggaaaccatt actccttgga acttcgagga agtggtcgat aagggcgcct
     3061 ctgcacagtc cttcatcgag agaatgacta attttgacaa aaacctgccc aatgagaaag
     3121 tgctgcctaa gcactccctg ctgtacgagt atttcactgt ctataacgaa ctgacaaagg
     3181 tgaaatacgt caccgagggc atgagaaagc cagccttcct gtcaggagag cagaagaaag
     3241 caatcgtgga tctgctgttt aaaaccaaca ggaaagtgac tgtcaagcag ctgaaggagg
     3301 actacttcaa gaaaattgaa tgcttcgatt ccgtggagat cagcggagtc gaagacagat
     3361 ttaacgcaag cctgggcgct taccacgatc tgctgaagat cattaaggat aaagacttcc
     3421 tggacaacga ggaaaatgag gatatcctgg aagacattgt gctgacactg accctgtttg
     3481 aggacagagg aatgatcgag gaaagactga aaacctatgc tcatctgttc gatgacaagg
     3541 tgatgaaaca gctgaagaga aggcgctaca ctggctgggg tagactgagc aggaagctga
     3601 tcaacggcat tagggataaa cagtcaggaa agacaatcct ggactttctg aaaagtgatg
     3661 gcttcgccaa ccgcaatttt atgcagctga ttcacgatga cagtctgacc ttcaaagagg
     3721 acatccagaa ggctcaggtg tctggacagg gccactccct gcatgagcag attgcaaacc
     3781 tggctggaag cccagccatc aagaaaggca ttctgcagac agtgaaaatc gtcgatgagc
     3841 tggtgaaagt catgggccat aagcccgaaa acatcgtgat tgagatggct cgcgaaaatc
     3901 agacaaccca gaagggtcag aagaacagta gagagaggat gaaaagaatc gaggaaggca
     3961 ttaaggagct gggtagccag atcctgaaag agcacccagt ggaaaacaca cagctgcaga
     4021 atgagaagct gtatctgtac tatctgcaga atggaagaga tatgtacgtg gaccaggagc
     4081 tggatattaa caggctgtct gattacgacg tggatcatat cgtcccccag agtttcatca
     4141 aagatgacag cattgacaac aaggtgctga ccaggtccga caaaaacaga ggaaaatcag
     4201 ataatgtccc tagtgaggaa gtggtcaaga aaatgaagaa ctactggaga cagctgctga
     4261 atgccaaact gatcactcag aggaagtttg ataacctgac aaaagcagag cgcggtggac
     4321 tgtcagaact ggacaaagct ggattcatca agaggcagct ggtggaaaca cgccagatca
     4381 ctaaacacgt cgcacagatt ctggatagtc gcatgaacac aaagtacgat gagaatgaca
     4441 aactgatcag agaagtgaag gtcattaccc tgaagagtaa actggtcagc gactttagga
     4501 aagatttcca gttttataag gtccgcgaga ttaacaatta tcaccatgcc catgacgcat
     4561 acctgaacgc cgtggtcggt accgcactga tcaagaaata cccaaaactg gagagcgaat
     4621 tcgtgtacgg agactataag gtgtacgatg tcagaaaaat gatcgccaag tccgagcagg
     4681 aaattggaaa agctactgcc aagtatttct tttactcaaa catcatgaat ttctttaaga
     4741 cagagatcac cctggccaat ggagaaatcc gcaaaaggcc cctgattgag acaaacggag
     4801 agacaggcga aatcgtgtgg gacaaaggca gagattttgc aaccgtgagg aaggtcctga
     4861 gcatgcctca agtgaatatc gtcaagaaaa ctgaggtgca gacaggcggt ttctcaaaag
     4921 aaagtattct gccaaaacgc aactctgata agctgatcgc tagaaagaaa gactgggacc
     4981 ctaagaagta tggaggcttt gactctccca ctgtggcata ctccgtcctg gtggtcgcta
     5041 aggtggagaa gggcaaaagc aagaaactga aatctgtcaa ggagctgctg ggtatcacaa
     5101 ttatggagag aagctctttc gagaagaacc caatcgattt tctggaggcc aaaggttata
     5161 aggaagtgaa gaaagacctg atcattaaac tgcccaagta cagtctgttt gagctggaaa
     5221 acggcaggaa acgcatgctg gcaagcgctg gagagctgca gaaaggcaat gaactggccc
     5281 tgccttctaa gtacgtgaac ttcctgtatc tggcaagcca ctacgagaag ctgaaaggat
     5341 ctccagagga taacgaacag aaacagctgt ttgtggagca gcacaagcat tatctggacg
     5401 agatcattga acagattagc gagttctcta aaagagtgat cctggccgac gcaaatctgg
     5461 ataaggtcct gtctgcttac aacaaacaca gagataagcc catcagggag caggccgaaa
     5521 atatcattca tctgttcact ctgacaaacc tgggcgcacc tgcagctttc aagtacttcg
     5581 acactacaat cgatagaaag aggtacacct ccactaagga ggtgctggac gctacactga
     5641 tccatcagag tattaccggc ctgtacgaaa caaggattga cctgtctcag ctgggtggcg
     5701 actcaccggg ggatccacta gtgggcagtg gagctactaa cttcagcctg ctgaagcagg
     5761 ctggtgacgt cgaggagaat cctggcccca tgaccgagta caagcccacg gtgcgcctcg
     5821 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     5881 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     5941 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     6001 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     6061 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     6121 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     6181 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     6241 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     6301 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     6361 ggtgcatgac ccgcaagccc ggtgcctgat tcgaaggcct gtcgtgaatt cactcctcag
     6421 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
     6481 cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     6541 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     6601 tgtctctcac tcggaaggac atatgctggc tgccatgaac tagctactcg ggacccctta
     6661 ccgaaacatc gccgcattct gcagaggagt cgagtgtatg taaacttctg acccactggg
     6721 aatgtgatga aagaaataaa agctgaaatg aatcattctc tctactatta ttctgatatt
     6781 tcacattctt aaaataaagt ggtgatccta actgacctaa gacagggaat ttttactagg
     6841 attaaatgtc aggaattgtg aaaaagtgag tttaaatgta tttggctaag gtgtatgtaa
     6901 acttccgact tcaactgtat agggatccgc ttcctcgctc actgactcgc tgcgctcggt
     6961 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     7021 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     7081 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     7141 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7201 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7261 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7321 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7381 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7441 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7501 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     7561 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7621 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7681 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7741 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7801 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7861 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7921 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7981 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     8041 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     8101 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     8161 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     8221 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     8281 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8341 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8401 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8461 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8521 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8581 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8641 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8701 gacacggaaa tgttgaatac tcat
//
