LOCUS       sequence-314451-        6876 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pMAGIC R4-R3 entry plasmid, contains 2xNLS SpCas9 (nuclease active) 
            for 3 or 4-component MultiSite Gateway Pro assembly..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6876)
  TITLE     Unpublished pMVP and pMAGIC plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6876)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: LP102: pMAGIC (R4-R3) NLS-Sp Cas9-NLS
FEATURES             Location/Qualifiers
     source          1..6876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      79..88
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             91..111
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             136..4236
                     /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             136..4236
                     /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             4237..4284
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    4292..4415
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4470..4489)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4471..4489)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4494..4510)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4494..4510)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4612..4631)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4623..5432
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4690..4709)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5579..6167
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6068..6087
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6321..6338
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6497..6524)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6616..6702)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     6751..6773
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6765..6782
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6766..6782
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(join(6832..6876,1..79))
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 tttttatgca aaatctaatt taatatattg atatttatat cattttacgt ttctcgttca
       61 acttttctat acaaagttgc caccatggcc ccaaagaaga agcggaaggt cggtatccac
      121 ggagtcccag cagccgacaa gaagtacagc atcggcctgg acatcggcac caactctgtg
      181 ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc
      241 aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc
      301 gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag
      361 aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc
      421 ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac
      481 cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac
      541 cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg
      601 gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc
      661 gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc
      721 gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg
      781 agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc
      841 ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc
      901 gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac
      961 aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg
     1021 tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc
     1081 ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa
     1141 gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag
     1201 aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc
     1261 aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag
     1321 gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg
     1381 ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac
     1441 cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc
     1501 aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg
     1561 aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc
     1621 aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag
     1681 tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag
     1741 cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac
     1801 cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac
     1861 tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat
     1921 ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg
     1981 gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg
     2041 aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac
     2101 accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc
     2161 aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg
     2221 atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag
     2281 ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc
     2341 atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc
     2401 gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac
     2461 agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg
     2521 aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg
     2581 cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac
     2641 gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg
     2701 ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg
     2761 aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag
     2821 ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc
     2881 atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac
     2941 tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc
     3001 accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc
     3061 gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc
     3121 ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac
     3181 gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac
     3241 ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag
     3301 atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag
     3361 ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa
     3421 aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc
     3481 gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc
     3541 cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa
     3601 ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag
     3661 aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc
     3721 aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct
     3781 gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg
     3841 tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag
     3901 ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc
     3961 tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag
     4021 caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc
     4081 aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac
     4141 accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac
     4201 gagacacgga tcgacctgtc tcagctggga ggcgacaaac ggccggcagc cacgaagaag
     4261 gccggccagg caaaaaagaa aaagggatcc gcaactttgt ataataaagt tgaacgagaa
     4321 acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa acagactaca
     4381 taatactgta aaacacaaca tatccagtca ctatgaatca actacttaga tggtattagt
     4441 gacctgtact gcagggcggc cgcgatatcc cctatagtga gtcgtattac atggtcatag
     4501 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     4561 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     4621 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     4681 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     4741 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     4801 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     4861 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     4921 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     4981 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     5041 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     5101 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     5161 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     5221 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     5281 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     5341 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     5401 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     5461 ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg
     5521 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5581 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5641 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5701 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5761 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5821 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5881 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5941 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6001 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6061 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6121 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6181 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6241 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     6301 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     6361 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     6421 ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag
     6481 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct
     6541 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa
     6601 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
     6661 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta
     6721 ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca
     6781 gtcttaagct cgggccccta caggtcacta ataccatcta agtagttgat tcatagtgac
     6841 tggatatgtt gtgttttaca gtattatgta gtctgt
//
