LOCUS       sequence-319767-        8892 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses 3xFLAG tagged hSpCas9 (Ae. aegypti PUb promoter) with 
            cloning site for sgRNAs (Ae. aegypti U6 promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8892)
  AUTHORS   Rozen-Gagnon K, Yi S, Jacobson E, Novack S, Rice CM
  TITLE     A selectable, plasmid-based system to generate CRISPR/Cas9 gene 
            edited and knock-in mosquito cell lines.
  JOURNAL   Sci Rep. 2021 Jan 12;11(1):736. doi: 10.1038/s41598-020-80436-5.
  PUBMED    33436886
REFERENCE   2  (bases 1 to 8892)
  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"; journalName: "Sci Rep."; 
            date: "2021-01-12"; volume: "11(1):736. doi"; pages: " 
            10.1038/s41598-020-80436-5"
COMMENT     Sequence Label: pKRG3-mU6-PUb-3xFLAG-hSpCas9
FEATURES             Location/Qualifiers
     source          1..8892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        194..269
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     regulatory      1840..1849
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1849..1914
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1921..1941
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1966..6066
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAKNIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6070..6090
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(6107..6131)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          6223..6288
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             6418..6438
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6863..6997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6900..6919)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6954..6973
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             join(7094..7162,7163..7954)
                     /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(7312..7331)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8125..8713
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8614..8633
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8867..8884
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gggttggccg attcattaat gcaggcaact
       61 cgtgaaaggt aggcggatca gcgaatgaaa tcgcccatcg agttgatacg tccatccatc
      121 gctagaaccg cgttcgctgt agaagactat ataagagcag aggcaagagt agtgaaatgg
      181 agacgacgtc tctgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
      241 aacttgaaaa agtggcaccg agtcggtgct tttttgtttt agagctagaa atagcaagtt
      301 aaaataaggc tagtccgttt ttagcgcttg cgccaattct gcagacaaat ggctatcttt
      361 acatgtagct tgtgcattga atccaattat aatttgcctt ggcaccagct gagccagaca
      421 agaaagaaag cttcccagaa gtatatcgat ttagaagggt tgacgtcact tgctgactgc
      481 actaatacag caaatgatgc aattagaatg attcaagtga aattcccaaa ttactgattt
      541 ttctctggat ttggttatca gattacattc gaagctaaga ttagctaccg aaattgtcga
      601 tcaaatcagg aaatcctttc tctatcgaaa aaggcattcg cacatcttcc tctctatgcc
      661 atatacacga agggtaggta cattgacgtc tttgccagaa gttgaactgc atcgttcaag
      721 gtacagaatg aacgactaac agacacaagc acgttttgct gtccattcag acacagggat
      781 ggtacccata ttcgatcgat atagagccat ccaaccgaac agaggtatat gtatgaatgt
      841 attgctgaaa ttttctagaa gtacaaccac cactacgaca gtgtctataa aacgcccctg
      901 caaaggcgaa accagctcaa tcgaatacgt ttcctagtgg agtgaacatt acgcggccca
      961 agtaagcagt gccagtgcaa gtgaagtgaa gtctctagtg aaaaagagtg atccaattag
     1021 ccagaggaga aaatttcaga gtgaacaaag ctttattcaa aggacaatta ctattaaatt
     1081 ggtgaaagtg catttcggtg aagggaatct tctagtgaag gtaggtaaat taattgatga
     1141 aattatagct atgagcgaaa actagtttgg tgaatgattc ctttgtcttt gaatgagcaa
     1201 actattttcc aagatggcga ctattgagct ttgagtgatt agtgaaaatt tgcaacgcag
     1261 tttcatcatc attgataaaa cccaattgtg attcacagcg ataatcatat ttcgttgaat
     1321 catcgctact aattgaatta aatttctaga ataataagaa taacgtattt gctccgtcac
     1381 atatctaaaa taaatatttt gatggtaatt acccattaag gtaatattaa cacatatcga
     1441 gaaaaacctt gaggaaatcg tgaaaacttg aagatacgca atttcaaaac tacgtagttc
     1501 aaagtcgaaa acaagttaat ttttcactta aaagtagggc gttgttgtga cgtcatcacc
     1561 ttcaagtgta tatttttcac ttggcctgcg actgcaaacg cagacaaagc aaaacaagtt
     1621 taaaacctgt cgtgtcgtgc tcgaagccaa aggcaatgaa tcaatatcaa atgagagttt
     1681 gcatttcaca accaattact caagcgtttc ctcgtttctt tttctgctca acagagattt
     1741 caacaccaag ctcgataagc ttgttcgaat ctcgagtgcg cgcttccgga ggtatacacc
     1801 taggcggtac cactgcagtg aattcggagc tctaccggtg ccaccatgga ctataaggac
     1861 cacgacggag actacaagga tcatgatatt gattacaaag acgatgacga taagatggcc
     1921 ccaaagaaga agcggaaggt cggtatccac ggagtcccag cagccgacaa gaagtacagc
     1981 atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag
     2041 gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac
     2101 ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga
     2161 accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc
     2221 agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg
     2281 gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg
     2341 gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc
     2401 gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc
     2461 cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc
     2521 cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg
     2581 gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc
     2641 gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg
     2701 ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg
     2761 agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac
     2821 gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg
     2881 agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac
     2941 gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag
     3001 tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga
     3061 gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc
     3121 gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac
     3181 aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag
     3241 gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc
     3301 cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc
     3361 agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct
     3421 tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag
     3481 gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa
     3541 gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag
     3601 gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag
     3661 gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg
     3721 ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc
     3781 ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt
     3841 gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa
     3901 gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg
     3961 atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac
     4021 ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag
     4081 gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat
     4141 ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag
     4201 ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag
     4261 aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag
     4321 ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg
     4381 cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag
     4441 gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt
     4501 ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag
     4561 agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg
     4621 ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc
     4681 ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag
     4741 atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat
     4801 gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc
     4861 cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac
     4921 gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc
     4981 gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag
     5041 caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc
     5101 aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac
     5161 ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg
     5221 ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc
     5281 aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg
     5341 gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg
     5401 gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc
     5461 accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc
     5521 tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg
     5581 gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg
     5641 gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag
     5701 ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg
     5761 gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat
     5821 ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc
     5881 aagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac
     5941 tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc
     6001 ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga
     6061 ggcgacagcc ccaagaagaa gagaaaggtg gaggccagct aataggaccc agctttcttg
     6121 tacaaagtgg tgacgtaagc tagcaggatc tttgtgaagg aaccttactt ctgtggtgtg
     6181 acataattgg acaaactacc tacagagatt taaagctcta aggtaaatat aaaattttta
     6241 agtgtataat gtgttaaact actgattcta attgtttgtg tattttagat tccaacctat
     6301 ggaactgatg aatgggagca gtggtggaat gcctttaatg aggaaaacct gttttgctca
     6361 gaagaaatgc catctagtga tgatgaggct actgctgact ctcaacattc tactcctcca
     6421 aaaaagaaga gaaaggtagt tgaccccaag gactttcctt cagaattgct aagttttttg
     6481 agtcatgctg tgtttagtaa tagaactctt gcttgctttg ctatttacac cacaaaggaa
     6541 aaagctgcac tgctatacaa gaaaattatg gaaaaatatt ctgtaacctt tataagtagg
     6601 cataacagtt ataatcataa catactgttt tttcttactc cacacaggca tagagtgtct
     6661 gctattaata actatgctca aaaattgtgt acctttagct ttttaatttg taaaggggtt
     6721 aataaggaat atttgatgta tagtgccttg actagagatc ataatcagcc ataccacatt
     6781 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     6841 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     6901 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6961 gtccaaactc atcaatgtat cttatcatgt ctggatcccg tttaaactac gcgtaattca
     7021 aacagggttc tggcgtcgtt ctcgtactgt tttccccagg ccagtgcttt agcgttattg
     7081 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     7141 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaagg atgctgaaga
     7201 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     7261 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     7321 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     7381 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     7441 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     7501 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     7561 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     7621 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     7681 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     7741 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     7801 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     7861 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     7921 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     7981 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     8041 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     8101 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     8161 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     8221 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     8281 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     8341 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     8401 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     8461 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     8521 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     8581 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     8641 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     8701 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     8761 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     8821 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     8881 cgaggacgaa ga
//
