LOCUS       sequence-171413-        8269 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses the ATP1A1 G2 sgRNA in combination with FLAGless 
            eSpCas9(1.1) to target ATP1A1 exon 4. Px330-like plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8269)
  AUTHORS   Agudelo D, Duringer A, Bozoyan L, Huard CC, Carter S, Loehr J, 
            Synodinou D, Drouin M, Salsman J, Dellaire G, Laganiere J, Doyon Y
  TITLE     Marker-free coselection for CRISPR-driven genome editing in human 
            cells.
  JOURNAL   Nat Methods. 2017 Apr 17. doi: 10.1038/nmeth.4265.
  PUBMED    28417998
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: "Nat 
            Methods. 2017 Apr 17. doi: 10.1038/nmeth.4265."
COMMENT     Sequence Label: eSpCas9(1.1)_No_FLAG_ATP1A1_G2
FEATURES             Location/Qualifiers
     source          1..8269
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          5..232
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      244..253
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             256..276
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(301..2838,2839..2841,2842..3303,3304..3306,3307..3474,
                     3475..3477,3478..4401)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Slaymaker et al., 2016)"
                     /label=eSpCas9(1.1)
                     /note="carries the mutations K848A, K1003A, and R1060A"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4402..4449
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4477..4494)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4483..4690
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4699..4839
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4699..4828
                     /label=AAV2 ITR
     rep_origin      4914..5369
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5001..5020)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5211..5232
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5386..5405)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5505..5527
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5565..5583)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5651..5755
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5756..5824,5825..6616)
                     /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     complement(5974..5993)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6787..7375
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7276..7295
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7437..7685
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7437..7457
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7608..7627
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7706..7781
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        7878..8163
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
       61 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
      121 tcctccgggc tgtaattagc tgagcaagag gtaagggttt aagggatggt tggttggtgg
      181 ggtattaatg tttaattacc tggagcacct gcctgaaatc actttttttc aggttggacc
      241 ggtgccacca tggccccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc
      301 gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc
      361 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
      421 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
      481 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
      541 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
      601 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
      661 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
      721 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
      781 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
      841 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
      901 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
      961 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg
     1021 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
     1081 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
     1141 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
     1201 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
     1261 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
     1321 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
     1381 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
     1441 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
     1501 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
     1561 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
     1621 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
     1681 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
     1741 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
     1801 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
     1861 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
     1921 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
     1981 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
     2041 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
     2101 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
     2161 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
     2221 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
     2281 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
     2341 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
     2401 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
     2461 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
     2521 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
     2581 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
     2641 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
     2701 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
     2761 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc
     2821 gtgcctcaga gctttctggc cgacgactcc atcgacaaca aggtgctgac cagaagcgac
     2881 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
     2941 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
     3001 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
     3061 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
     3121 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
     3181 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
     3241 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
     3301 cctgcgctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
     3361 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
     3421 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaaggcgcct
     3481 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc
     3541 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
     3601 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc
     3661 agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat
     3721 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
     3781 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
     3841 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
     3901 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag
     3961 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
     4021 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
     4081 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
     4141 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc
     4201 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
     4261 gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag
     4321 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
     4381 ctgtctcagc tgggaggcga caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa
     4441 aagaaaaagt aagaattcct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     4501 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     4561 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     4621 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagag aatagcaggc
     4681 atgctgggga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     4741 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     4801 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     4861 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     4921 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4981 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5041 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     5101 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     5161 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     5221 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     5281 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5341 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     5401 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5461 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5521 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     5581 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     5641 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5701 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5761 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5821 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5881 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5941 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6001 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6061 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6121 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6181 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6241 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6301 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6361 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6421 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg gaagccgcgg
     6481 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6541 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6601 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6661 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6721 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6781 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6841 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6901 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6961 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7021 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7081 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7141 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7201 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7261 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7321 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7381 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtgagg
     7441 gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt agagagataa
     7501 ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt
     7561 aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc
     7621 ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa
     7681 acaccgatcc aagctgctac agaaggtttt agagctagaa atagcaagtt aaaataaggc
     7741 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgtt ttagagctag
     7801 aaatagcaag ttaaaataag gctagtccgt ttttagcgcg tgcgccaatt ctgcagacaa
     7861 atggctctag aggtacccgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     7921 aacgaccccc gcccattgac gtcaatagta acgccaatag ggactttcca ttgacgtcaa
     7981 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     8041 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcattg tgcccagtac
     8101 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     8161 atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc
     8221 ccaattttgt atttatttat tttttaatta ttttgtgcag cgatggggg
//
