LOCUS       sequence-256476-        8332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  U6 promoter expresses customizable Spyo-guide; EF1a promoter 
            provides puromycin resistance. This vector is a derivative of the 
            lentiGuide vector, with minor modifications to the tracrR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8332)
  AUTHORS   DeWeirdt PC, Sangree AK, Hanna RE, Sanson KR, Hegde M, Strand C, 
            Persky NS, Doench JG
  TITLE     Genetic screens in isogenic mammalian cell lines without single cell
            cloning.
  JOURNAL   Nat Commun. 2020 Feb 6;11(1):752. doi: 10.1038/s41467-020-14620-6.
  PUBMED    32029722
REFERENCE   2  (bases 1 to 8332)
  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 
            Commun."; date: "2020-02-6"; volume: "11(1):752. doi"; pages: " 
            10.1038/s41467-020-14620-6"
COMMENT     Sequence Label: pRDA_118
FEATURES             Location/Qualifiers
     source          1..8332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    83..104
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(119..124,125..142,143..149)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    157..173
                     /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     162..184
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     181..197
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     181..197
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     216..236
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        218..236
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        264..490
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             491..671
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    718..843
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1336..1569
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1754..1798
                     /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        1979..2227
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1979..1999
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2150..2169
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2249..2324
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2386..2503
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2642..3820
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2873..3811
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3768..3788
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3833..4432
                     /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(3833..3852)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4329..4349
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4448..5036
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4501..4521)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4919..4930)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5108..5341
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5419..5540
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5456..5475)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5510..5529
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5580..5715
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5735..5754)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5736..5754)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5764..5781)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5764..5780)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5773..5795)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5922..6377
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6009..6028)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6219..6240
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6403..6507
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6508..6576,6577..7368)
                     /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(6726..6745)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7539..8127
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8028..8047
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8281..8298
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
       61 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      121 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      181 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
      241 aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat
      301 ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt
      361 ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg
      421 gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg
      481 atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
      541 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
      601 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
      661 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc
      721 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg
      781 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc
      841 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
      901 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
      961 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
     1021 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
     1081 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
     1141 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc
     1201 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
     1261 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
     1321 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
     1381 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
     1441 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
     1501 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
     1561 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg
     1621 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga
     1681 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
     1741 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg
     1801 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg
     1861 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca
     1921 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccggtaccga
     1981 gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat
     2041 aattagaatt aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa
     2101 gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat
     2161 gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg
     2221 aaacaccgga gacgcttttt tcgtctcagt ttgagagcta gaaatagcaa gttcaaataa
     2281 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt gaattcaagc
     2341 ttggcgtaac tagatcttga gacaaatggc agtattcatc cacaatttta aaagaaaagg
     2401 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2461 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     2521 cagcagagat ccactttggc gccggctcga gggggcccgg gtgcaaagat ggataaagtt
     2581 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
     2641 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
     2701 gggaggggtc ggcaattgat ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
     2761 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     2821 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
     2881 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
     2941 tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg
     3001 ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg
     3061 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc
     3121 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg
     3181 gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc
     3241 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
     3301 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
     3361 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
     3421 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag
     3481 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc
     3541 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
     3601 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     3661 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     3721 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     3781 agtggttcaa agtttttttc ttccatttca ggtgtcgtga cgtacggcca ccatgaccga
     3841 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     3901 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     3961 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     4021 ggtgtgggtc gcggacgacg gcgccgccgt ggcggtctgg accacgccgg agagcgtcga
     4081 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     4141 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     4201 gttcctggcc accgtcggag tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     4261 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     4321 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     4381 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaacgcgtta
     4441 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4501 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4561 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4621 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4681 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4741 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4801 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     4861 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4921 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4981 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg
     5041 actttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     5101 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
     5161 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     5221 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     5281 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     5341 agtacgtata gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     5401 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5461 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5521 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
     5581 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     5641 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     5701 gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg
     5761 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     5821 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     5881 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
     5941 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     6001 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     6061 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     6121 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     6181 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     6241 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     6301 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     6361 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     6421 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6481 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6541 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6601 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6661 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6721 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6781 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6841 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6901 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6961 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     7021 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     7081 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     7141 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     7201 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     7261 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     7321 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     7381 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7441 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7501 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7561 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7621 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7681 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7741 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7801 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7861 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7921 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7981 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     8041 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     8101 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     8161 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     8221 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     8281 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tc
//
