Nme2sgRNA_pLKO

LOCUS       Nme2sgRNA_pLKO                9023 bp ds-DNA     circular SYN 18-JAN-2019
DEFINITION  Nme2Cas9 U6-driven sgRNA cassette.
ACCESSION   .
VERSION     .
KEYWORDS    Nme2sgRNA_pLKO
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9023)
  AUTHORS   Edraki A, Mir A, Ibraheim R, Gainetdinov I, Yoon Y, Song CQ, Cao Y, 
            Gallant J, Xue W, Rivera-Perez JA, Sontheimer EJ
  TITLE     A Compact, High-Accuracy Cas9 with a Dinucleotide PAM for In Vivo 
            Genome Editing.
  JOURNAL   Mol Cell. 2018 Dec 18. pii: S1097-2765(18)31033-5. doi: 
            10.1016/j.molcel.2018.12.003.
  PUBMED    30581144
REFERENCE   2  (bases 1 to 9023)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 18, 2019 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9023
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(168..756)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(248..267)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(927..1787)
                     /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     1550..1569
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1788..1892)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1918..2373)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2055..2076)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2267..2286
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2500..2522
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2514..2531
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2515..2531
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2541..2560
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2541..2559
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      2580..2715
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2634..2653)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2755..2876
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2766..2785)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2820..2839
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             2948..3181
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     CDS             complement(3309..3908)
                     /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(3392..3412)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3889..3908
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(3930..4440)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3993..4011)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     misc_feature    4489..4606
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        complement(4655..4747)
                     /label=Nm tracrRNA
                     /note="trans-activating CRISPR RNA for the Neisseria 
                     meningitidis CRISPR/Cas9 system (Hou et al., 2013)"
     CDS             4940..5734
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4994..5013)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5604..5623
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     promoter        complement(5873..6113)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     complement(5923..5942)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(6093..6113)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    6640..6873
                     /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    7366..7491
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             7538..7718
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(7719..7945)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(7973..7993)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7973..7991)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8012..8028)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8012..8028)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8025..8047)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8036..8052
                     /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(8060..8089)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     mobile_element  8090..8857
                     /mobile_element_type="insertion sequence:IS1"
                     /note="prokaryotic transposable element"
     protein_bind    8882..8903
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9020..14)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
       61 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
      121 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
      181 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      241 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      301 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      361 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      421 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      481 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
      541 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      601 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      661 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
      721 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
      781 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
      841 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
      901 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
      961 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     1021 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     1081 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1141 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1201 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1261 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     1321 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     1381 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     1441 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     1501 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     1561 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     1621 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     1681 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     1741 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     1801 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     1861 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     1921 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     1981 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     2041 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     2101 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     2161 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     2221 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     2281 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     2341 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
     2401 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     2461 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     2521 cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggt acgtccctag
     2581 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
     2641 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     2701 agttaggggc gggatgggcg gagttagggg cgggatagct agagccagac atgataagat
     2761 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     2821 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttcctc
     2881 tcactctctg atattcattt ctttgcaagt tataaatact gaataataag atgacatgaa
     2941 ctactactgc tagagatttt ccacactgac taaaagggtc tgagggatct ctagttacca
     3001 gagtcacaca acagacgggc acacactact tgaagcactc aaggcaagct ttattgaggc
     3061 ttaagcagtg ggttccctag ttagccagag agctcccagg ctcagatctg gtctaaccag
     3121 agagacccag tacaagcaaa aagcagatct tgtcttcgtt gggagtgaat tagcccttcc
     3181 agtcccccct tttcttttaa aaagtggcta agatctacag ctgccttgta agtcattggt
     3241 cttaaaggta ccgatgcatg gggtcgtgcg ctcctttcgg tcgggcgctg cgggtcgtgg
     3301 ggcgggcgtc aggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc
     3361 tcgacgtcgg cggtgacggt gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg
     3421 gaggtctcca ggaaggcggg caccccggcg cgctcggccg cctccactcc ggggagcacg
     3481 acggcgctgc ccagaccctt gccctggtgg tcgggcgaga cgccgacggt ggccaggaac
     3541 cacgcgggct ccttgggccg gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc
     3601 agccgggaac cgctcaactc ggccatgcgc gggccgatct cggcgaacac cgcccccgct
     3661 tcgacgctct ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc
     3721 ttgccgatgt cgagcccgac gcgcgtgagg aagagttctt gcagctcggt gacccgctcg
     3781 atgtggcggt ccggatcgac ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg
     3841 agggtgcgta cggccctggg gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac
     3901 tcggtcatgg taagctccgg tggatccccc tggggagaga ggtcggtgat tcggtcaacg
     3961 agggagccga ctgccgacgt gcgctccgga ggcttgcaga atgcggaaca ccgcgcgggc
     4021 aggaacaggg cccacactac cgccccacac cccgcctccc gcaccgcccc ttcccggccg
     4081 ctgctctcgg cgcgccctgc tgagcagccg ctattggcca cagcccatcg cggtcggcgc
     4141 gctgccattg ctccctggcg ctgtccgtct gcgagggtac tagtgagacg tgcggcttcc
     4201 gtttgtcacg tccggcacgc cgcgaaccgc aaggaacctt cccgacttag gggcggagca
     4261 ggaagcgtcg ccggggggcc cacaagggta gcggcgaaga tccgggtgac gctgcgaacg
     4321 gacgtgaaga atgtgcgaga cccagggtcg gcgccgctgc gtttcccgga accacgccca
     4381 gagcagccgc gtccctgcgc aaacccaggg ctgccttgga aaaggcgcaa ccccaacccc
     4441 ctcgagccgc ggccaaagtg gatctctgct gtccctgtaa taaacccgaa aattttgaat
     4501 ttttgtaatt tgtttttgta attctttagt ttgtatgtct gttgctatta tgtctactat
     4561 tctttcccct gcactgtacc ccccaatccc cccttttctt ttaaaattgt ggatgaatac
     4621 tgccatttgt ctcgaggtcg agaattcaaa aaaataaacg atgcccctta aagcagaagc
     4681 tttaaggggc agagcgttgc ggcacatctt ttcagacggc cttattgtag caacggttct
     4741 catttcgttt ccgaaatgag aaagggagct acaacgtcag caggttctgc tagctgatcc
     4801 gtcggaattg ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg
     4861 gatggctttc ttgccgccaa ggatctgatg gcgcagggga tcaagatctg atcaagagac
     4921 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4981 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     5041 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     5101 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     5161 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     5221 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5281 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5341 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5401 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5461 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5521 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5581 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5641 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5701 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgagagctcg
     5761 cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga
     5821 acgctcggtt gccgccgggc gtttggatca ctcacctgca caccggtgtt tcgtcctttc
     5881 cacaagatat ataaagccaa gaaatcgaaa tactttcaag ttacggtaag catatgatag
     5941 tccattttaa aacataattt taaaactgca aactacccaa gaaattatta ctttctacgt
     6001 cacgtatttt gtactaatat ctttgtgttt acagtcaaat taattccaat tatctctcta
     6061 acagccttgt atcgtatatg caaatatgaa ggaatcatgg gaaataggcc ctcggtgaag
     6121 ggggcggccg ctcgaggcta gtctcgtgat cgataccgtc gagatccgtt cactaatcga
     6181 atggatctgt ctctgtctct ctctccacct tcttcttcta ttccttcggg cctgtcgggt
     6241 cccctcgggg ttgggaggtg ggtctgaaac gataatggtg aatatccctg cctaactcta
     6301 ttcactatag aaagtacagc aaaaactatt cttaaaccta ccaagcctcc tactatcatt
     6361 atgaataatt ttatatacca cagccaattt gttatgttaa accaattcca caaacttgcc
     6421 catttatcta attccaataa ttcttgttca ttcttttctt gctggttttg cgattcttca
     6481 attaaggagt gtattaagct tgtgtaattg ttaatttctc tgtcccactc catccaggtc
     6541 gtgtgattcc aaatctgttc cagagattta ttactccaac tagcattcca aggcacagca
     6601 gtggtgcaaa tgagttttcc agagcaaccc caaatcccca ggagctgttg atcctttagg
     6661 tatctttcca cagccaggat tcttgcctgg agctgcttga tgccccagac tgtgagttgc
     6721 aacagatgct gttgcgcctc aatagccctc agcaaattgt tctgctgctg cactatacca
     6781 gacaataatt gtctggcctg taccgtcagc gtcattgacg ctgcgcccat agtgcttcct
     6841 gctgctccca agaacccaag gaacaaagct cctattccca ctgctctttt ttctctctgc
     6901 accactcttc tctttgcctt ggtgggtgct actcctaatg gttcaatttt tactacttta
     6961 tatttatata attcacttct ccaattgtcc ctcatatctc ctcctccagg tctgaagatc
     7021 agcggccgct tgctgtgcgg tggtcttact tttgttttgc tcttcctcta tcttgtctaa
     7081 agcttccttg gtgtctttta tctctatcct ttgatgcaca caatagaggg ttgctactgt
     7141 attatataat gatctaagtt cttctgatcc tgtctgaagg gatggttgta gctgtcccag
     7201 tatttgtcta cagccttctg atgtttctaa caggccagga ttaactgcga atcgttctag
     7261 ctccctgctt gcccatacta tatgttttaa tttatatttt ttctttcccc ctggccttaa
     7321 ccgaattttt tcccatcgcg atctaattct cccccgctta atactgacgc tctcgcaccc
     7381 atctctctcc ttctagcctc cgctagtcaa aatttttggc gtactcacca gtcgccgccc
     7441 ctcgcctctt gccgtgcgcg cttcagcaag ccgagtcctg cgtcgagaga gctcctctgg
     7501 tttccctttc gctttcaagt ccctgttcgg gcgccactgc tagagatttt ccacactgac
     7561 taaaagggtc tgagggatct ctagttacca gagtcacaca acagacgggc acacactact
     7621 tgaagcactc aaggcaagct ttattgaggc ttaagcagtg ggttccctag ttagccagag
     7681 agctcccagg ctcagatctg gtctaaccag agagacccgt ttatgtatcg agctaggcac
     7741 ttaaatacaa tatctctgca atgcggcaat tcagtggttc gtccaatcca tgtcagaccc
     7801 gtctgttgcc ttcctaataa ggcacgatcg taccacctta cttccaccaa tcggcatgca
     7861 cggtgctttt tctctccttg taaggcatgt tgctaactca tcgttaccat gttgcaagac
     7921 tacaagagta ttgcataaga ctacattaag cttgcagctc cagcttttgt tccctttagt
     7981 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     8041 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaag gtgatgctgc
     8101 caacttactg atttagtgta tgatggtgtt tttgaggtgc tccagtggct tctgtttcta
     8161 tcagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg
     8221 gacatcagcg ctatctctgc tctcactgcc gtaaaacatg gcaactgcag ttcacttaca
     8281 ccgcttctca acccggtacg caccagaaaa tcattgatat ggccatgaat ggcgttggat
     8341 gccgggcaac agcccgcatt atgggcgttg gcctcaacac gattttacgt cacttaaaaa
     8401 actcaggccg cagtcggtaa cctcgcgcat acagccgggc agtgacgtca tcgtctgcgc
     8461 ggaaatggac gaacagtggg gctatgtcgg ggctaaatcg cgccagcgct ggctgtttta
     8521 cgcgtatgac agtctccgga agacggttgt tgcgcacgta ttcggtgaac gcactatggc
     8581 gacgctgggg cgtcttatga gcctgctgtc accctttgac gtggtgatat ggatgacgga
     8641 tggctggccg ctgtatgaat cccgcctgaa gggaaagctg cacgtaatca gcaagcgata
     8701 tacgcagcga attgagcggc ataacctgaa tctgaggcag cacctggcac ggctgggacg
     8761 gaagtcgctg tcgttctcaa aatcggtgga gctgcatgac aaagtcatcg ggcattatct
     8821 gaacataaaa cactatcaat aagttggagt cattaccaaa gtgtaaagcc tggggtgcct
     8881 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     8941 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     9001 ttgggcgctc ttccgcttcc tcg
//