LOCUS       pLH-sgRNA1-2XMS2                8327 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  sgRNA1-2XMS2.
ACCESSION   .
VERSION     .
KEYWORDS    pLH-sgRNA1-2XMS2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8327)
  AUTHORS   Ma H, Tu LC, Naseri A, Huisman M, Zhang S, Grunwald D, Pederson T
  TITLE     Multiplexed labeling of genomic loci with dCas9 and engineered 
            sgRNAs using CRISPRainbow.
  JOURNAL   Nat Biotechnol. 2016 Apr 18. doi: 10.1038/nbt.3526.
  PUBMED    27088723
REFERENCE   2  (bases 1 to 8327)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            //www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8327
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(167..187)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    714..947
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1440..1565
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             1612..1792
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(1793..2019)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(2047..2067)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2047..2065)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2086..2102)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2086..2102)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2099..2121)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2110..2126
                     /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)."
     promoter        complement(2134..2164)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2179..2200
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2317..2334)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2488..3076)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2568..2587)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3247..4107)
                     /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     3870..3889
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4108..4212)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4238..4693)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4375..4396)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4587..4606
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4820..4842
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4834..4851
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4835..4851
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4861..4880
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4861..4879
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      4900..5035
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4954..4973)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5075..5196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5086..5105)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5140..5159
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             5268..5501
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     5583..5599
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5584..5600
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(5624..6649)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(6739..7249)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6802..6820)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     misc_feature    7298..7415
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        complement(7494..7512)
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        complement(7564..7582)
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     CDS             complement(7620..7925)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(7722..7741)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     promoter        complement(8274..187)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     complement(8324..16)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 taagcatatg atagtccatt ttaaaacata attttaaaac tgcaaactac ccaagaaatt
       61 attactttct acgtcacgta ttttgtacta atatctttgt gtttacagtc aaattaattc
      121 caattatctc tctaacagcc ttgtatcgta tatgcaaata tgaaggaatc atgggaaata
      181 ggccctcggt gaagggggcg gccgctcgag gctagtctcg tgatcgatac cgtcgagatc
      241 cgttcactaa tcgaatggat ctgtctctgt ctctctctcc accttcttct tctattcctt
      301 cgggcctgtc gggtcccctc ggggttggga ggtgggtctg aaacgataat ggtgaatatc
      361 cctgcctaac tctattcact atagaaagta cagcaaaaac tattcttaaa cctaccaagc
      421 ctcctactat cattatgaat aattttatat accacagcca atttgttatg ttaaaccaat
      481 tccacaaact tgcccattta tctaattcca ataattcttg ttcattcttt tcttgctggt
      541 tttgcgattc ttcaattaag gagtgtatta agcttgtgta attgttaatt tctctgtccc
      601 actccatcca ggtcgtgtga ttccaaatct gttccagaga tttattactc caactagcat
      661 tccaaggcac agcagtggtg caaatgagtt ttccagagca accccaaatc cccaggagct
      721 gttgatcctt taggtatctt tccacagcca ggattcttgc ctggagctgc ttgatgcccc
      781 agactgtgag ttgcaacaga tgctgttgcg cctcaatagc cctcagcaaa ttgttctgct
      841 gctgcactat accagacaat aattgtctgg cctgtaccgt cagcgtcatt gacgctgcgc
      901 ccatagtgct tcctgctgct cccaagaacc caaggaacaa agctcctatt cccactgctc
      961 ttttttctct ctgcaccact cttctctttg ccttggtggg tgctactcct aatggttcaa
     1021 tttttactac tttatattta tataattcac ttctccaatt gtccctcata tctcctcctc
     1081 caggtctgaa gatcagcggc cgcttgctgt gcggtggtct tacttttgtt ttgctcttcc
     1141 tctatcttgt ctaaagcttc cttggtgtct tttatctcta tcctttgatg cacacaatag
     1201 agggttgcta ctgtattata taatgatcta agttcttctg atcctgtctg aagggatggt
     1261 tgtagctgtc ccagtatttg tctacagcct tctgatgttt ctaacaggcc aggattaact
     1321 gcgaatcgtt ctagctccct gcttgcccat actatatgtt ttaatttata ttttttcttt
     1381 ccccctggcc ttaaccgaat tttttcccat cgcgatctaa ttctcccccg cttaatactg
     1441 acgctctcgc acccatctct ctccttctag cctccgctag tcaaaatttt tggcgtactc
     1501 accagtcgcc gcccctcgcc tcttgccgtg cgcgcttcag caagccgagt cctgcgtcga
     1561 gagagctcct ctggtttccc tttcgctttc aagtccctgt tcgggcgcca ctgctagaga
     1621 ttttccacac tgactaaaag ggtctgaggg atctctagtt accagagtca cacaacagac
     1681 gggcacacac tacttgaagc actcaaggca agctttattg aggcttaagc agtgggttcc
     1741 ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac ccgtttatgt
     1801 atcgagctag gcacttaaat acaatatctc tgcaatgcgg caattcagtg gttcgtccaa
     1861 tccatgtcag acccgtctgt tgccttccta ataaggcacg atcgtaccac cttacttcca
     1921 ccaatcggca tgcacggtgc tttttctctc cttgtaaggc atgttgctaa ctcatcgtta
     1981 ccatgttgca agactacaag agtattgcat aagactacat taagcttgca gctccagctt
     2041 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     2101 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     2161 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2221 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2281 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2341 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2401 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2461 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2521 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2581 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2641 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2701 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2761 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2821 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2881 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2941 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3001 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3061 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3121 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3181 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3241 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3301 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3361 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3421 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3481 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3541 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3601 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3661 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3721 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3781 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3841 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     3901 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3961 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4021 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4081 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4141 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4201 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     4261 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     4321 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     4381 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     4441 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     4501 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     4561 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     4621 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     4681 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     4741 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     4801 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg
     4861 taatacgact cactataggg cgaattgggt acgtccctag gcctccaaaa aagcctcctc
     4921 actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa
     4981 ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg
     5041 gagttagggg cgggatagct agagccagac atgataagat acattgatga gtttggacaa
     5101 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     5161 ttatttgtaa ccattataag ctgcaataaa caagttcctc tcactctctg atattcattt
     5221 ctttgcaagt tataaatact gaataataag atgacatgaa ctactactgc tagagatttt
     5281 ccacactgac taaaagggtc tgagggatct ctagttacca gagtcacaca acagacgggc
     5341 acacactact tgaagcactc aaggcaagct ttattgaggc ttaagcagtg ggttccctag
     5401 ttagccagag agctcccagg ctcagatctg gtctaaccag agagacccag tacaagcaaa
     5461 aagcagatct tttcttcgtt gggagtgaat tagcccttcc agtcccccct tttcttttaa
     5521 aaagtggcta agatctacag ctgccttgta agtcattggt cttaaaggta ccgggccccc
     5581 cctcgaggtc gacggtatcg atagatcccg gtcggcatct actctattcc tttgccctcg
     5641 gacgagtgct ggggcgtcgg tttccactat cggcgagtac ttctacacag ccatcggtcc
     5701 agacggccgc gcttctgcgg gcgatttgtg tacgcccgac agtcccggct ccggatcgga
     5761 cgattgcgtc gcatcgaccc tgcgcccaag ctgcatcatc gaaattgccg tcaaccaagc
     5821 tctgatagag ttggtcaaga ccaatgcgga gcatatacgc ccggagccgc ggcgatcctg
     5881 caagctccgg atgcctccgc tcgaagtagc gcgtctgctg ctccatacaa gccaaccacg
     5941 gcctccagaa gaagatgttg gcgacctcgt attgggaatc cccgaacatc gcctcgctcc
     6001 agtcaatgac cgctgttatg cggccattgt ccgtcaggac attgttggag ccgaaatccg
     6061 cgtgcacgag gtgccggact tcggggcagt cctcggccca aagcatcagc tcatcgagag
     6121 cctgcgcgac ggacgcactg acggtgtcgt ccatcacagt ttgccagtga tacacatggg
     6181 gatcagcaat cgcgcatatg aaatcacgcc atgtagtgta ttgaccgatt ccttgcggtc
     6241 cgaatgggcc gaacccgctc gtctggctaa gatcggccgc agcgatcgca tccatggcct
     6301 ccgcgaccgg ctgcagaaca gcgggcagtt cggtttcagg cagatcttgc aacgtgacac
     6361 cctgtgcacg gcgggagatg caataggtca ggctctcgct aaattcccca atgtcaagca
     6421 cttccggaat cgggagcgcg gccgatgcaa agtgccgata aacataacga tctttgtaga
     6481 aaccatcggc gcagctattt acccgcagga catatccacg ccctcctaca tcgaagctga
     6541 aagcacgaga ttcttcgccc tccgagagct gcatcaggtc ggagacgctg tcgaactttt
     6601 cgatcagaaa cttctcgaca gacgtcgcgg tgagttcagg ctttttcata tctcattgcc
     6661 ccccgggatc agatctgcag ggcttcccaa ccttaccaga gggcgcccca gctggcaagc
     6721 ttgatccggt ggatccccct ggggagagag gtcggtgatt cggtcaacga gggagccgac
     6781 tgccgacgtg cgctccggag gcttgcagaa tgcggaacac cgcgcgggca ggaacagggc
     6841 ccacactacc gccccacacc ccgcctcccg caccgcccct tcccggccgc tgctctcggc
     6901 gcgccctgct gagcagccgc tattggccac agcccatcgc ggtcggcgcg ctgccattgc
     6961 tccctggcgc tgtccgtctg cgagggtact agtgagacgt gcggcttccg tttgtcacgt
     7021 ccggcacgcc gcgaaccgca aggaaccttc ccgacttagg ggcggagcag gaagcgtcgc
     7081 cggggggccc acaagggtag cggcgaagat ccgggtgacg ctgcgaacgg acgtgaagaa
     7141 tgtgcgagac ccagggtcgg cgccgctgcg tttcccggaa ccacgcccag agcagccgcg
     7201 tccctgcgca aacccagggc tgccttggaa aaggcgcaac cccaaccccc tcgagccgcg
     7261 gccaaagtgg atctctgctg tccctgtaat aaacccgaaa attttgaatt tttgtaattt
     7321 gtttttgtaa ttctttagtt tgtatgtctg ttgctattat gtctactatt ctttcccctg
     7381 cactgtaccc cccaatcccc ccttttcttt taaaattgtg gatgaatact gccatttgtc
     7441 tcgaggtcga gaattcaaaa aaagcaccga ctcggtgcca cttggccctg cagacatggg
     7501 tgatcctcat gttggccaag ttgataacgg actagcctta tttgaacttg ctaggccctg
     7561 cagacatggg tgatcctcat gttggcctag ctctcaaacg ggtcttcgtc ctgcagggtt
     7621 tatattcccc agaacatcag gttaatggcg tttttgatgt cattttcgcg gtggctgaga
     7681 tcagccactt cttccccgat aacggagacc ggcacactgg ccatatcggt ggtcatcatg
     7741 cgccagcttt catccccgat atgcaccacc gggtaaagtt cacgggagac tttatctgac
     7801 agcagacgtg cactggccag ggggatcacc atccgtcgcc cgggcgtgtc aataatatca
     7861 ctctgtacat ccacaaacag acgataacgg ctctctcttt tataggtgta aaccttaaac
     7921 tgcatttcac cagtccctgt tctcgtcagc aaaagagccg ttcatttcaa taaaccgggc
     7981 gacctcagcc atcccttcct gattttccgc tttccagcgt tcggcacgca gacgacgggc
     8041 ttcattctgc atggttgtgc ttaccagacc ggagatattg acatcatata tgccttgagc
     8101 aactgatagc tgtcgctgtc aactgtcact gtaatacgct gcttcatagc acacctcttt
     8161 ttgacatact tcgggtatac atatcagtat atattcttat accgcaaaaa tcagcgcgca
     8221 aatacgcata ctgttatctg gcttttagta agcctcgaag acaccggtgt ttcgtccttt
     8281 ccacaagata tataaagcca agaaatcgaa atactttcaa gttacgg
//