LOCUS       sequence-340312-        7136 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of Cas9-T2A-mClover3 synthetic construct .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7136)
  TITLE     Verstreken lab plasmids
REFERENCE   2  (bases 1 to 7136)
  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"
COMMENT     Sequence Label: pIVTR(T7A)-Cas9-T2A-mClover3
FEATURES             Location/Qualifiers
     source          1..7136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      102..111
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             111..176
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             183..203
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             228..4328
                     /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
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             228..4328
                     /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
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4329..4376
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4392..4445
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(4446..4448,4449..5162)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     5096..5117
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     rep_origin      complement(5381..5969)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5461..5480)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6140..6931,6932..7000))
                     /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     6763..6782
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7001..7105)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taatacgact cactataagg acatttgctt ctgacacaac tgtgttcact agcaacctca
       61 aacagaattc gagctcggta cccggggatc ctctagagtc tgccaccatg gactataagg
      121 accacgacgg agactacaag gatcatgata ttgattacaa agacgatgac gataagatgg
      181 ccccaaagaa gaagcggaag gtcggtatcc acggagtccc agcagccgac aagaagtaca
      241 gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca
      301 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga
      361 acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga
      421 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct
      481 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc
      541 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg
      601 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca
      661 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg
      721 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca
      781 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg
      841 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga
      901 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc
      961 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc
     1021 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt
     1081 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc
     1141 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg
     1201 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga
     1261 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg
     1321 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca
     1381 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg
     1441 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc
     1501 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct
     1561 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga
     1621 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg
     1681 cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga
     1741 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca
     1801 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa
     1861 aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag
     1921 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc
     1981 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact
     2041 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt
     2101 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca
     2161 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc
     2221 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg
     2281 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag
     2341 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca
     2401 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg
     2461 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag
     2521 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag
     2581 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc
     2641 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc
     2701 aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct
     2761 ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca
     2821 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc
     2881 tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag
     2941 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc
     3001 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga
     3061 atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
     3121 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg
     3181 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa
     3241 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg
     3301 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt
     3361 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa
     3421 acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag
     3481 tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca
     3541 gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact
     3601 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg
     3661 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga
     3721 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg
     3781 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc
     3841 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac
     3901 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga
     3961 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc
     4021 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta
     4081 atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg
     4141 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt
     4201 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca
     4261 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg
     4321 gaggcgacaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag aaaaaggaat
     4381 tcggcagtgg agagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg
     4441 gcccagtgtc taagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     4501 acggcgacgt aaacggccac aagttcagcg tccgcggcga gggcgagggc gatgccacca
     4561 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     4621 ccctcgtgac caccttcggc tacggcgtgg cctgcttcag ccgctacccc gaccacatga
     4681 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     4741 ctttcaagga cgacggtacc tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     4801 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     4861 acaagctgga gtacaacttc aacagccact acgtctatat cacggccgac aagcagaaga
     4921 actgcatcaa ggctaacttc aagatccgcc acaacgttga ggacggcagc gtgcagctcg
     4981 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     5041 actacctgag ccatcagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     5101 tcctgctgga gttcgtgacc gccgccggga ttacacatgg catggacgag ctgtacaagt
     5161 aagacctgca ggcatgcaag cttgctcgct ttcttgctgt ccaatttcta ttaaaggttc
     5221 ctttgttccc taagtccaac tactaaactg ggggatatta tgaagggcct tgagcatctg
     5281 gattctgcct aataaaaaac atttattttc attgcaaaac catgggagca aaaggccagc
     5341 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5401 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5461 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5521 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5581 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5641 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5701 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5761 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5821 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5881 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5941 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6001 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6061 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6121 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6181 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6241 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6301 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6361 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6421 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6481 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6541 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6601 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6661 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6721 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6781 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6841 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6901 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6961 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7021 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7081 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
