LOCUS       sequence-473535-        8323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lentiviral gRNA plasmid targeting human PIK3R5.
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: PIK3R5 gRNA (BRDN0001146461)
FEATURES             Location/Qualifiers
     source          1..8323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    148..273
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             320..500
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(501..727)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(755..775)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(755..773)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(794..810)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(794..810)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(807..829)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    818..834
                     /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(join(842..848,849..866,867..872))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    887..908
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1025..1042)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1196..1784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1276..1295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1955..2746,2747..2815))
                     /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     2578..2597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2816..2920)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2946..3401)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3083..3104)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3295..3314
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3528..3550
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3542..3559
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3543..3559
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3569..3588
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3569..3587
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      3608..3743
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3783..3904
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3794..3813)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3848..3867
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             3982..4215
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    4287..4875
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             4393..4404
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4802..4822
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4891..5490)
                     /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(4974..4994)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5471..5490
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(5503..6681)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5512..6450
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(5535..5555)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    6820..6937
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        complement(6993..7068)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(7090..7338)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(7148..7167)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(7318..7338)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(7516..7560)
                     /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"
     misc_feature    7745..7978
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 tgatgtttct aacaggccag gattaactgc gaatcgttct agctccctgc ttgcccatac
       61 tatatgtttt aatttatatt ttttctttcc ccctggcctt aaccgaattt tttcccatcg
      121 cgatctaatt ctcccccgct taatactgac gctctcgcac ccatctctct ccttctagcc
      181 tccgctagtc aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg
      241 cgcttcagca agccgagtcc tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa
      301 gtccctgttc gggcgccact gctagagatt ttccacactg actaaaaggg tctgagggat
      361 ctctagttac cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag
      421 ctttattgag gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc
      481 tggtctaacc agagagaccc gtttatgtat cgagctaggc acttaaatac aatatctctg
      541 caatgcggca attcagtggt tcgtccaatc catgtcagac ccgtctgttg ccttcctaat
      601 aaggcacgat cgtaccacct tacttccacc aatcggcatg cacggtgctt tttctctcct
      661 tgtaaggcat gttgctaact catcgttacc atgttgcaag actacaagag tattgcataa
      721 gactacatta agcttgcagc tccagctttt gttcccttta gtgagggtta attgcgcgct
      781 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
      841 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
      901 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
      961 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     1021 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     1081 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     1141 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1201 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1261 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1321 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1381 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1441 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1501 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     1561 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     1621 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     1681 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1741 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1801 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     1861 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     1921 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     1981 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     2041 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     2101 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     2161 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     2221 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     2281 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     2341 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     2401 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     2461 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     2521 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     2581 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     2641 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     2701 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     2761 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     2821 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     2881 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     2941 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3001 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3061 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3121 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3181 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3241 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3301 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3361 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
     3421 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     3481 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     3541 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac
     3601 gtccctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg
     3661 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
     3721 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggatagctag agccagacat
     3781 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     3841 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     3901 agttcctctc actctctgat attcatttct ttgcaagtta taaatactga ataataagat
     3961 gacatgaact actatacgta ctgctagaga ttttccacac tgactaaaag ggtctgaggg
     4021 atctctagtt accagagtca cacaacagac gggcacacac tacttgaagc actcaaggca
     4081 agctttattg aggcttaagc agtgggttcc ctagttagcc agagagctcc caggctcaga
     4141 tctggtctaa ccagagagac ccagtacaag caaaaagcag atcttgtctt cgttgggagt
     4201 gaattagccc ttccagtccc cccttttctt ttaaaaagtg gctaagatct acagctgcct
     4261 tgtaagtcat tggtcttaaa gtcgacgcgg ggaggcggcc caaagggaga tccgactcgt
     4321 ctgagggcga aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg
     4381 aaggtccgct ggattgaggg ccgaagggac gtagcagaag gacgtcccgc gcagaatcca
     4441 ggtggcaaca caggcgagca gccaaggaaa ggacgatgat ttccccgaca acaccacgga
     4501 attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag
     4561 ttccgccgtg gcaataggga gggggaaagc gaaagtcccg gaaaggagct gacaggtggt
     4621 ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca gtgcacacca cgccacgttg
     4681 cctgacaacg ggccacaact cctcataaag agacagcaac caggatttat acaaggagga
     4741 gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc
     4801 acatagcgta aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta
     4861 atccagaggt tgattgtcga cttaacgcgt tcaggcaccg ggcttgcggg tcatgcacca
     4921 ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta
     4981 gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc
     5041 cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga
     5101 gactccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc
     5161 ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc gcgggccgat
     5221 ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccacggcggc
     5281 gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc
     5341 ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg acggtgtggc gcgtggcggg
     5401 gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt cgcgggtggc
     5461 gaggcgcacc gtgggcttgt actcggtcat ggtggccgta cgtcacgaca cctgaaatgg
     5521 aagaaaaaaa ctttgaacca ctgtctgagg cttgagaatg aaccaagatc caaactcaaa
     5581 aagggcaaat tccaaggaga attacatcaa gtgccaagct ggcctaactt cagtctccac
     5641 ccactcagtg tggggaaact ccatcgcata aaacccctcc ccccaaccta aagacgacgt
     5701 actccaaaag ctcgagaact aatcgaggtg cctggacggc gcccggtact ccgtggagtc
     5761 acatgaagcg acggctgagg acggaaaggc ccttttcctt tgtgtgggtg actcacccgc
     5821 ccgctctccc gagcgccgcg tcctccattt tgagctccct gcagcagggc cgggaagcgg
     5881 ccatctttcc gctcacgcaa ctggtgccga ccgggccagc cttgccgccc agggcggggc
     5941 gatacacggc ggcgcgaggc caggcaccag agcaggccgg ccagcttgag actacccccg
     6001 tccgattctc ggtggccgcg ctcgcaggcc ccgcctcgcc gaacatgtgc gctgggacgc
     6061 acgggccccg tcgccgcccg cggccccaaa aaccgaaata ccagtgtgca gatcttggcc
     6121 cgcatttaca agactatctt gccagaaaaa aagcgtcgca gcaggtcatc aaaaatttta
     6181 aatggctaga gacttatcga aagcagcgag acaggcgcga aggtgccacc agattcgcac
     6241 gcggcggccc cagcgcccag gccaggcctc aactcaagca cgaggcgaag gggctcctta
     6301 agcgcaaggc ctcgaactct cccacccact tccaacccga agctcgggat caagaatcac
     6361 gtactgcagc caggtggaag taattcaagg cacgcaaggg ccataacccg taaagaggcc
     6421 aggcccgcgg gaaccacaca cggcacttac ctgtgttctg gcggcaaacc cgttgcgaaa
     6481 aagaacgttc acggcgacta ctgcacttat atacggttct cccccaccct cgggaaaaag
     6541 gcggagccag tacacgacat cactttccca gtttaccccg cgccaccttc tctaggcacc
     6601 ggatcaattg ccgacccctc cccccaactt ctcggggact gtgggcgatg tgcgctctgc
     6661 ccactgacgg gcaccggagc caattcccac tcctttcaag acctagaagg tccattagct
     6721 gcaaagattc ctctctgttt aaaactttat ccatctttgc acccgggccc cctcgagccg
     6781 gcgccaaagt ggatctctgc tgtccctgta ataaacccga aaattttgaa tttttgtaat
     6841 ttgtttttgt aattctttag tttgtatgtc tgttgctatt atgtctacta ttctttcccc
     6901 tgcactgtac cccccaatcc ccccttttct tttaaaattg tggatgaata ctgccatttg
     6961 tctcaagatc tagttacgcc aagcttaaaa aagcaccgac tcggtgccac tttttcaagt
     7021 tgataacgga ctagccttat tttaacttgc tatttctagc tctaaaacat gacttcgctt
     7081 cacaggcacg gtgtttcgtc ctttccacaa gatatataaa gccaagaaat cgaaatactt
     7141 tcaagttacg gtaagcatat gatagtccat tttaaaacat aattttaaaa ctgcaaacta
     7201 cccaagaaat tattactttc tacgtcacgt attttgtact aatatctttg tgtttacagt
     7261 caaattaatt ctaattatct ctctaacagc cttgtatcgt atatgcaaat atgaaggaat
     7321 catgggaaat aggccctctc tgggtcccct cggggttggg aggtgggtct gaaacgataa
     7381 tggtgaatat ccctgcctaa ctctattcac tatagaaagt acagcaaaaa ctattcttaa
     7441 acctaccaag cctcctacta tcattatgaa taattttata taccacagcc aatttgttat
     7501 gttaaaccaa ttccacaaac ttgcccattt atctaattcc aataattctt gttcattctt
     7561 ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt aagcttgtgt aattgttaat
     7621 ttctctgtcc cactccatcc aggtcgtgtg attccaaatc tgttccagag atttattact
     7681 ccaactagca ttccaaggca cagcagtggt gcaaatgagt tttccagagc aaccccaaat
     7741 ccccaggagc tgttgatcct ttaggtatct ttccacagcc aggattcttg cctggagctg
     7801 cttgatgccc cagactgtga gttgcaacag atgctgttgc gcctcaatag ccctcagcaa
     7861 attgttctgc tgctgcacta taccagacaa taattgtctg gcctgtaccg tcagcgtcat
     7921 tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac ccaaggaaca aagctcctat
     7981 tcccactgct cttttttctc tctgcaccac tcttctcttt gccttggtgg gtgctactcc
     8041 taatggttca atttttacta ctttatattt atataattca cttctccaat tgtccctcat
     8101 atctcctcct ccaggtctga agatcagcgg ccgcttgctg tgcggtggtc ttacttttgt
     8161 tttgctcttc ctctatcttg tctaaagctt ccttggtgtc ttttatctct atcctttgat
     8221 gcacacaata gagggttgct actgtattat ataatgatct aagttcttct gatcctgtct
     8281 gaagggatgg ttgtagctgt cccagtattt gtctacagcc ttc
//
