pPUR-mU6-sgRNA-2XPP7

LOCUS       pPUR-mU6-sgRNA-2XPP7            8026 bp DNA     circular SYN 09-MAY-2026
DEFINITION  Empty backbone for sgRNA-2XPP7 in CRIPSR FISHer system.
ACCESSION   .
VERSION     .
KEYWORDS    pPUR-mU6-sgRNA-2XPP7
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8026)
  AUTHORS   Lyu XY, Deng Y, Huang XY, Li ZZ, Fang GQ, Yang D, Wang FL, Kang W, 
            Shen EZ, Song CQ
  TITLE     CRISPR FISHer enables high-sensitivity imaging of nonrepetitive DNA 
            in living cells through phase separation-mediated signal 
            amplification.
  JOURNAL   Cell Res. 2022 Nov;32(11):969-981. doi: 10.1038/s41422-022-00712-z. 
            Epub 2022 Sep 14.
  PUBMED    36104507
REFERENCE   2  (bases 1 to 8026)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 15, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41422-022-00712-z"; journalName: "Cell Res"; date: 
            "2022-11"; volume: "32"; issue: "11"; pages: "969-981"
FEATURES             Location/Qualifiers
     source          1..8026
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        52..562
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     481..499
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             577..1173
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(577..596)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1073..1093
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1183..1239
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1255..1965
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGIYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             1966..1986
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     LTR             2082..2315
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2387..2508
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2424..2443)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2478..2497
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2548..2683
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2703..2722)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2704..2722)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2732..2749)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2732..2748)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2741..2763)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2890..3345
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2977..2996)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3187..3208
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3371..3475
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3476..4336
                     /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(3694..3713)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4507..5095
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4996..5015
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5249..5266
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5383..5404
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5419..5449
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5457..5473
                     /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)."
     primer_bind     5462..5484
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5481..5497
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5481..5497
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5516..5536
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5518..5536
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5564..5790
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5791..5971
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6018..6143
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6636..6869
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7054..7098
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        7399..7712
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     7631..7651
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     misc_feature    join(7912..8026,1..3)
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 tttcgggttt attacaggga cagcagagat ccactttggc cgcggctcga gggggttggg
       61 gttgcgcctt ttccaaggca gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg
      121 ttccgggaaa cgcagcggcg ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag
      181 cgtcacccgg atcttcgccg ctacccttgt gggccccccg gcgacgcttc ctgctccgcc
      241 cctaagtcgg gaaggttcct tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc
      301 acgtctcact agtaccctcg cagacggaca gcgccaggga gcaatggcag cgcgccgacc
      361 gcgatgggct gtggccaata gcggctgctc agcagggcgc gccgagagca gcggccggga
      421 aggggcggtg cgggaggcgg ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg
      481 gtgttccgca ttctgcaagc ctccggagcg cacgtcggca gtcggctccc tcgttgaccg
      541 aatcaccgac ctctctcccc agggggatcc gccaccatga ccgagtacaa gcccacggtg
      601 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
      661 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
      721 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
      781 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
      841 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
      901 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
      961 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     1021 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     1081 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     1141 cgcacctggt gcatgacccg caagcccggt gccgctagcg gtgctactaa cttcagcctg
     1201 ctgaagcagg ctggagacgt ggaggagaac cctggacctg gtagtggaac gcgtatggtg
     1261 tctaagggcg aagagctgat taaggagaac atgcacatga agctgtacat ggagggcacc
     1321 gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
     1381 cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
     1441 gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg catccccgac
     1501 ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaagat
     1561 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
     1621 gtcaagatca gaggggtgaa cttcacatcc aacggccctg tgatgcagaa gaaaacactc
     1681 ggctgggagg ccttcaccga gacgctgtac cccgctgacg gcggcctgga aggcagaaac
     1741 gacatggccc tgaagctcgt gggcgggagc catctgatcg caaacgccaa gaccacatat
     1801 agatccaaga aacccgctaa gaacctcaag atgcctggca tctactatgt ggactacaga
     1861 ctggaaagaa tcaaggaggc caacaacgag acctacgtcg agcagcacga ggtggcagtg
     1921 gccagatact gcgacctccc tagcaaactg gggcacaagc ttaatccaaa gaaaaagcgg
     1981 aaagtgtgag tcgacctcga gggggggccc ggtaccttta agaccaatga cttacaaggc
     2041 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     2101 ccaacgaaga aaagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
     2161 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     2221 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     2281 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
     2341 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     2401 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     2461 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     2521 ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     2581 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     2641 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt
     2701 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     2761 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     2821 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     2881 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2941 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3001 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3061 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3121 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3181 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3241 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3301 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     3361 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3421 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3481 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3541 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3601 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3661 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3721 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3781 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3841 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3901 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3961 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4021 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4081 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4141 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4201 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4261 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4321 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4381 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4441 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4501 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4561 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4621 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4681 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4741 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4801 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4861 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4921 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4981 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5041 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5101 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5161 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     5221 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     5281 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     5341 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     5401 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     5461 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
     5521 taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata
     5581 ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa
     5641 aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg
     5701 caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt
     5761 gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc
     5821 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     5881 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     5941 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
     6001 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
     6061 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
     6121 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
     6181 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
     6241 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
     6301 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
     6361 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
     6421 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
     6481 cggccactga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
     6541 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
     6601 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
     6661 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
     6721 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
     6781 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
     6841 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
     6901 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
     6961 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
     7021 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
     7081 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
     7141 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
     7201 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
     7261 ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac
     7321 agatccattc gattagtgaa cggatctcga cggtatcgat cacgagacta gcctcgagcg
     7381 gccgccctgt gctctagaga tccgacgccg ccatctctag gcccgcgccg gccccctcgc
     7441 acagacttgt gggagaagct cggctactcc cctgccccgg ttaatttgca tataatattt
     7501 cctagtaact atagaggctt aatgtgcgat aaaagacaga taatctgttc tttttaatac
     7561 tagctacatt ttacatgata ggcttggatt tctataagag atacaaatac taaattatta
     7621 ttttaaaaaa cagcacaaaa ggaaactcac cctaactgta aagtaattgt gtgttttgag
     7681 actataaata tcccttggag aaaagccttg tttggtgtct tcgaacgcag gacgaagacc
     7741 cgtttgagag ctaccggagc agacgatatg gcgtcgctcc ggtagcaagt tcaaataagg
     7801 ctagtccgtt atcaacttgg agcagacgat atggcgtcgc tccaagtggc accgagtcgg
     7861 tgcttttttt gaattctcga cctcgagaca aatggcagta ttcatccaca attttaaaag
     7921 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     7981 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaat
//