LOCUS       sequence-152930-        8323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lentiviral gRNA plasmid targeting human MAP3K4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8323)
  AUTHORS   Doench JG, Fusi N, Sullender M, Hegde M, Vaimberg EW, Donovan KF, 
            Smith I, Tothova Z, Wilen C, Orchard R, Virgin HW, Listgarten J, 
            Root DE
  TITLE     Optimized sgRNA design to maximize activity and minimize off-target 
            effects of CRISPR-Cas9.
  JOURNAL   Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437.
  PUBMED    26780180
REFERENCE   2  (bases 1 to 8323)
  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 
            Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437."
COMMENT     Sequence Label: MAP3K4 gRNA (BRDN0001148639)
FEATURES             Location/Qualifiers
     source          1..8323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     75..94
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     328..345
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    462..483
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(498..503,504..521,522..528)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    536..552
                     /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     541..563
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     560..576
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     560..576
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     595..615
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        597..615
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        643..869
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             870..1050
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1097..1222
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1715..1948
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2133..2177
                     /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        2355..2603
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2355..2375
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2526..2545
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2625..2700
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2756..2873
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3012..4190
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3243..4181
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4138..4158
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4203..4802
                     /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(4203..4222)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4699..4719
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4818..5406
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4871..4891)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5289..5300)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5478..5711
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5789..5910
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5826..5845)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5880..5899
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5950..6085
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6105..6124)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6106..6124)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6134..6151)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6134..6150)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6143..6165)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6292..6747
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6379..6398)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6589..6610
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6773..6877
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6878..6946,6947..7738)
                     /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(7096..7115)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7909..8323,1..174)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
       61 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
      121 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
      181 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
      241 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
      301 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
      361 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
      421 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
      481 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
      541 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
      601 aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac
      661 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
      721 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
      781 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
      841 tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct
      901 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
      961 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     1021 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
     1081 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     1141 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     1201 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     1261 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     1321 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     1381 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     1441 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     1501 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     1561 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     1621 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     1681 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     1741 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     1801 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     1861 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     1921 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     1981 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     2041 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     2101 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     2161 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     2221 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     2281 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     2341 cgaggggacc cagagagggc ctatttccca tgattccttc atatttgcat atacgataca
     2401 aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa
     2461 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
     2521 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
     2581 tcttgtggaa aggacgaaac accgacataa tctgaacccc acatgtttta gagctagaaa
     2641 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
     2701 ttttttaagc ttggcgtaac tagatcttga gacaaatggc agtattcatc cacaatttta
     2761 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa
     2821 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt
     2881 attacaggga cagcagagat ccactttggc gccggctcga gggggcccgg gtgcaaagat
     2941 ggataaagtt ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag
     3001 gagtgggaat tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
     3061 agaagttggg gggaggggtc ggcaattgat ccggtgccta gagaaggtgg cgcggggtaa
     3121 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
     3181 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
     3241 aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg
     3301 tgccttgaat tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg
     3361 gaagtgggtg ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag
     3421 ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct
     3481 gtctcgctgc tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc
     3541 tttttttctg gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg
     3601 tttttggggc cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc
     3661 ggggcctgcg agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg
     3721 ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc
     3781 ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct
     3841 caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa
     3901 gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca
     3961 ggcacctcga ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt
     4021 tttatgcgat ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc
     4081 acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca
     4141 agcctcagac agtggttcaa agtttttttc ttccatttca ggtgtcgtga cgtacggcca
     4201 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     4261 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     4321 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     4381 acatcggcaa ggtgtgggtc gcggacgacg gcgccgccgt ggcggtctgg accacgccgg
     4441 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     4501 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     4561 agcccgcgtg gttcctggcc accgtcggag tctcgcccga ccaccagggc aagggtctgg
     4621 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     4681 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     4741 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     4801 gaacgcgtta agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     4861 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     4921 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     4981 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     5041 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     5101 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     5161 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     5221 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     5281 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     5341 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     5401 ccccgcgtcg actttaagac caatgactta caaggcagct gtagatctta gccacttttt
     5461 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt
     5521 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     5581 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     5641 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     5701 aatctctagc agtacgtata gtagttcatg tcatcttatt attcagtatt tataacttgc
     5761 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     5821 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5881 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     5941 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     6001 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     6061 tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcgt
     6121 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     6181 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     6241 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     6301 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     6361 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     6421 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     6481 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     6541 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     6601 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     6661 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     6721 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     6781 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6841 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6901 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6961 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     7021 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     7081 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     7141 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     7201 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     7261 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     7321 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     7381 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     7441 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     7501 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     7561 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     7621 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     7681 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7741 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7801 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7861 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7921 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7981 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     8041 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     8101 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     8161 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     8221 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     8281 gagataccta cagcgtgagc tatgagaaag cgccacgctt ccc
//
