LOCUS       sequence-211316-        8245 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  Mammalian expression vector for Streptococcus thermophilus CRISPR1 
            (St1Cas9 LMD-9) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8245)
  AUTHORS   Agudelo D, Carter S, Velimirovic M, Duringer A, Rivest JF, Levesque 
            S, Loehr J, Mouchiroud M, Cyr D, Waters PJ, Laplante M, Moineau S, 
            Goulet A, Doyon Y
  TITLE     Versatile and robust genome editing with Streptococcus thermophilus 
            CRISPR1-Cas9.
  JOURNAL   Genome Res. 2020 Jan;30(1):107-117. doi: 10.1101/gr.255414.119. Epub
            2020 Jan 3.
  PUBMED    31900288
REFERENCE   2  (bases 1 to 8245)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1101/gr.255414.119"; journalName: "Genome Res"; date: "2020-01";
            volume: "30"; issue: "1"; pages: "107-117"
COMMENT     Sequence Label: MSP1594_2x_NLS
FEATURES             Location/Qualifiers
     source          1..8245
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          117..1133
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1057..1079
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1141..1160
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1226..1235
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1241..1261
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1286..4645
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus 
                     thermophilus Type II CRISPR/Cas system (Esvelt et al., 
                     2013)"
                     /label=St1Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="SDLVLGLDIGIGSVGVGILNKVTGEIIHKNSRIFPAAQAENNLVR
                     RTNRQGRRLARRKKHRRVRLNRLFEESGLITDFTKISINLNPYQLRVKGLTDELSNEEL
                     FIALKNMVKHRGISYLDDASDDGNSSVGDYAQIVKENSKQLETKTPGQIQLERYQTYGQ
                     LRGDFTVEKDGKKHRLINVFPTSAYRSEALRILQTQQEFNPQITDEFINRYLEILTGKR
                     KYYHGPGNEKSRTDYGRYRTSGETLDNIFGILIGKCTFYPDEFRAAKASYTAQEFNLLN
                     DLNNLTVPTETKKLSKEQKNQIINYVKNEKAMGPAKLFKYIAKLLSCDVADIKGYRIDK
                     SGKAEIHTFEAYRKMKTLETLDIEQMDRETLDKLAYVLTLNTEREGIQEALEHEFADGS
                     FSQKQVDELVQFRKANSSIFGKGWHNFSVKLMMELIPELYETSEEQMTILTRLGKQKTT
                     SSSNKTKYIDEKLLTEEIYNPVVAKSVRQAIKIVNAAIKEYGDFDNIVIEMARETNEDD
                     EKKAIQKIQKANKDEKDAAMLKAANQYNGKAELPHSVFHGHKQLATKIRLWHQQGERCL
                     YTGKTISIHDLINNSNQFEVDHILPLSITFDDSLANKVLVYATANQEKGQRTPYQALDS
                     MDDAWSFRELKAFVRESKTLSNKKKEYLLTEEDISKFDVRKKFIERNLVDTRYASRVVL
                     NALQEHFRAHKIDTKVSVVRGQFTSQLRRHWGIEKTRDTYHHHAVDALIIAASSQLNLW
                     KKQKNTLVSYSEDQLLDIETGELISDDEYKESVFKAPYQHFVDTLKSKEFEDSILFSYQ
                     VDSKFNRKISDATIYATRQAKVGKDKADETYVLGKIKDIYTQDGYDAFMKIYKKDKSKF
                     LMYRHDPQTFEKVIEPILENYPNKQINEKGKEVPCNPFLKYKEEHGYIRKYSKKGNGPE
                     IKSLKYYDSKLGNHIDITPKDSNNKVVLQSVSPWRADVYFNKTTGKYEILGLKYADLQF
                     EKGTGTYKISQEKYNDIKKKEGVDSDSEFKFTLYKNDLLLVKDTETKEQQLFRFLSRTM
                     PKQKHYVELKPYDKQKFEGGEALIKVLGNVANSGQCKKGLGKSNISIYKVRTDVLGNQH
                     IIKNEGDKPKLDF"
     CDS             4658..4678
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4767..4786)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4832..4887
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4886..4905)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5248..5264)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5248..5264)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5261..5283)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5272..5288
                     /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(5296..5302,5303..5320,5321..5326))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5341..5362
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5421..5616
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5467..5602
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5622..5756
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5659..5678)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5713..5732
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5824..5841)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5995..6583)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6075..6094)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6754..7545,7546..7614))
                     /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     7377..7396
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7615..7719)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        7750..8129
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
       61 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
      121 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
      181 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
      241 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
      301 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt
      361 gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg
      421 ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg
      481 gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
      541 tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc
      601 ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg
      661 cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc
      721 cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct
      781 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
      841 gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct
      901 ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc
      961 ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg
     1021 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1081 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     1141 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattctgc agtcgacggt
     1201 accgcgggcc cgggatccac cggtcgccac catgggcgcc ccaaagaaga agcggaaggt
     1261 cggtatccac ggagtcccag cagccagcga cctggtgctg ggcctggaca tcggcatcgg
     1321 cagcgtgggc gtgggcatcc tgaacaaggt gaccggcgag atcatccaca agaacagtcg
     1381 catcttccct gctgctcagg ctgagaacaa cctggtgcgc cgcaccaacc gccagggtcg
     1441 ccggcttgct cgccgcaaga agcaccggcg cgtgcgcctg aaccgcctgt tcgaggagag
     1501 cggcctgatc accgacttca ccaagatcag catcaacctg aacccctacc agctgcgcgt
     1561 gaagggcctg accgacgagc tgagcaacga ggagctgttc atcgccctga agaacatggt
     1621 gaagcaccgc ggcatcagct acctggacga cgccagcgac gacggcaaca gcagcgtggg
     1681 cgactacgcc cagatcgtga aggagaacag caagcagctg gagaccaaga cccccggcca
     1741 gatccagctg gagcgctacc agacctacgg ccagctgcgc ggcgacttca ccgtggagaa
     1801 ggacggcaag aagcaccgcc tgatcaacgt gttccccacc agcgcctacc gcagcgaggc
     1861 cctgcgcatc ctgcagaccc agcaggagtt caacccccag atcaccgacg agttcatcaa
     1921 ccgctacctg gagatcctga ccggcaagcg caagtactac cacggccccg gcaacgagaa
     1981 gagccgcacc gactacggcc gctaccgcac cagcggcgag accctggaca acatcttcgg
     2041 catcctgatc ggcaagtgca ccttctaccc cgacgagttc cgcgccgcca aggccagcta
     2101 caccgcccag gagttcaatc tgctgaacga cctgaacaac ctgaccgtgc ccaccgagac
     2161 caagaagctg agcaaggagc agaagaacca gatcatcaac tacgtgaaga acgagaaggc
     2221 tatgggcccc gccaagctgt tcaagtacat cgccaagctg ctgagctgcg acgtggccga
     2281 catcaagggc taccgcatcg acaagagcgg caaggccgag atccacacct tcgaggccta
     2341 ccgcaagatg aagaccctgg agaccctgga catcgagcag atggaccgag agaccctgga
     2401 caagctggcc tacgtgctga ccctgaacac cgagcgcgag ggcatccagg aggccctgga
     2461 gcacgagttc gccgacggca gcttcagcca gaaacaggtg gacgagctgg tgcagttccg
     2521 caaggccaac agcagcatct tcggcaaggg ctggcacaac ttcagcgtga agctgatgat
     2581 ggagctgatc cccgagctgt acgagaccag cgaggagcag atgaccatcc tgacccgcct
     2641 gggcaagcag aagaccacca gcagcagcaa caagaccaag tacatcgacg agaagctgct
     2701 gaccgaggag atctacaacc ccgtggtggc caagagcgtg cgccaggcca tcaagatcgt
     2761 gaacgccgcc atcaaggagt acggcgactt cgacaacatc gtgatcgaga tggcccgcga
     2821 gaccaacgag gacgacgaga agaaggccat ccagaagatc cagaaggcca acaaggacga
     2881 gaaggacgcc gccatgctga aggccgccaa ccagtacaac ggcaaggccg agctgcccca
     2941 cagcgtgttc cacggccaca agcagctggc caccaagatc cgcctgtggc accagcaggg
     3001 cgagcgctgc ctgtacaccg gcaagaccat cagcatccac gacctgatca acaacagcaa
     3061 ccagttcgag gtggaccaca tcctgcccct gagcatcacc ttcgacgaca gcctggccaa
     3121 caaggtgctg gtgtacgcca ccgccaacca ggagaagggc cagcgcaccc cctaccaggc
     3181 cctggacagc atggacgacg cctggagctt ccgcgagctg aaggccttcg tgcgcgagag
     3241 caagaccctg agcaacaaga agaaggagta tctgctgacc gaggaggaca tcagcaagtt
     3301 cgacgtgcgc aagaagttca tcgagcgcaa cctggtggac acccgctacg ccagccgcgt
     3361 ggtgctgaac gccctgcagg agcacttccg cgcccacaag atcgacacca aggtgagcgt
     3421 ggtgcgcggc cagttcacca gccagctgcg ccgccactgg ggcatcgaga agacccgcga
     3481 cacctaccac caccacgccg tggacgccct gatcattgcg gcttctagcc agctgaacct
     3541 gtggaagaag cagaagaaca ccctggtgag ctacagcgag gaccagctgc tggacatcga
     3601 gaccggcgag ctgatcagcg acgacgagta caaggagagc gtgttcaagg ccccctacca
     3661 gcacttcgtg gacaccctga agagcaagga gttcgaggac agcatcctgt tcagctacca
     3721 ggtggacagc aagttcaacc gcaagatcag cgacgccacc atctacgcca cccgccaggc
     3781 caaggtgggc aaggacaagg ccgacgagac ctacgtgctg ggcaagatca aggacatcta
     3841 cacccaggac ggctacgacg ccttcatgaa gatctacaag aaggacaaga gcaagttcct
     3901 gatgtaccgc cacgaccccc agaccttcga gaaggtgatc gagcccatcc tggagaacta
     3961 ccccaacaag cagatcaacg agaaaggcaa ggaggtgccc tgcaacccct tcctgaagta
     4021 caaggaggag cacggctaca tccgcaagta cagcaagaag ggcaacggcc ccgagatcaa
     4081 gagcctgaag tactacgaca gcaagctggg caaccacatc gacatcaccc ccaaggacag
     4141 caacaacaag gtggtgctgc agagcgtgag cccctggcgc gccgacgtgt acttcaacaa
     4201 gaccaccggc aagtacgaga tcctggggct gaagtacgcc gatctgcagt ttgagaaagg
     4261 cacaggcacc tacaagatca gccaggagaa gtacaacgac atcaagaaga aggagggcgt
     4321 ggacagcgac agcgagttca agttcaccct gtacaagaac gaccttctgc tggtgaagga
     4381 caccgagacc aaggagcaac agctgttccg cttcctgagc cgcaccatgc ccaagcagaa
     4441 gcactacgtg gagctgaagc cctacgacaa gcagaagttc gagggcggcg aggccctgat
     4501 caaggtgctg ggcaacgtgg ccaacagcgg ccagtgcaag aagggcctgg gcaagagcaa
     4561 catcagcatc tacaaggtgc gcaccgacgt gctgggcaac cagcacatca tcaagaacga
     4621 gggcgacaag cccaagttgg acttcagcag ggctgacccc aagaagaaga ggaaggtgtg
     4681 agcggccgca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
     4741 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
     4801 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
     4861 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
     4921 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
     4981 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
     5041 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
     5101 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
     5161 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac
     5221 ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5281 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5341 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5401 cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5461 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5521 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     5581 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt
     5641 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5701 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct
     5761 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5821 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5881 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5941 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6001 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6061 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6121 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6181 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6241 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6301 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6361 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6421 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6481 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6541 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6601 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6661 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6721 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6781 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6841 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6901 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6961 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7021 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7081 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7141 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7201 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7261 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7321 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7381 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7441 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7501 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7561 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7621 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7681 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7741 acctgggtcg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     7801 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     7861 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     7921 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     7981 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     8041 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     8101 tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc
     8161 tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt
     8221 gtgcagcgat gggggcgggg ggggg
//
