LOCUS       sequence-343498-        8323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral gRNA vector targeting human CD73.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8323)
  AUTHORS   Wilk A, Hayat F, Cunningham R, Li J, Garavaglia S, Zamani L, 
            Ferraris DM, Sykora P, Andrews J, Clark J, Davis A, Chaloin L, Rizzi
            M, Migaud M, Sobol RW
  TITLE     Extracellular NAD(+) enhances PARP-dependent DNA repair capacity 
            independently of CD73 activity.
  JOURNAL   Sci Rep. 2020 Jan 20;10(1):651. doi: 10.1038/s41598-020-57506-9.
  PUBMED    31959836
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: "Sci Rep."; 
            date: "2020-01-20"; volume: "10(1):651. doi"; pages: " 
            10.1038/s41598-020-57506-9"
COMMENT     Sequence Label: pLentiGuide-Puro/CD73-gRNA4
FEATURES             Location/Qualifiers
     source          1..8323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        126..1304
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          357..1295
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1252..1272
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1317..1916
                     /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(1317..1336)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1813..1833
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    1932..2520
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1985..2005)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2403..2414)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2592..2825
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2903..3024
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2940..2959)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2994..3013
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3064..3199
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3219..3238)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3220..3238)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3248..3265)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3248..3264)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3257..3279)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3406..3861
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3493..3512)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3703..3724
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3887..3991
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3992..4060,4061..4852)
                     /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(4210..4229)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5023..5611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5512..5531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5765..5782
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5899..5920
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5935..5940,5941..5958,5959..5965)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5973..5989
                     /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     5978..6000
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5997..6013
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5997..6013
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6032..6052
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6034..6052
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6080..6306
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6307..6487
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6534..6659
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7152..7385
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7570..7614
                     /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        7792..8040
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7792..7812
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7963..7982
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8062..8137
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    8193..8310
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gggacagcag agatccactt tggcgccggc tcgagggggc ccgggtgcaa agatggataa
       61 agttttaaac agagaggaat ctttgcagct aatggacctt ctaggtcttg aaaggagtgg
      121 gaattggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
      181 tggggggagg ggtcggcaat tgatccggtg cctagagaag gtggcgcggg gtaaactggg
      241 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
      301 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa
      361 gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt
      421 gaattacttc cacctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg
      481 ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg
      541 cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg
      601 ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt
      661 tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg
      721 gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc
      781 tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg
      841 tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg
      901 caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat
      961 ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct
     1021 ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc
     1081 tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg
     1141 cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga
     1201 tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc
     1261 agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgacgtacg gccaccatga
     1321 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
     1381 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
     1441 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     1501 gcaaggtgtg ggtcgcggac gacggcgccg ccgtggcggt ctggaccacg ccggagagcg
     1561 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     1621 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     1681 cgtggttcct ggccaccgtc ggagtctcgc ccgaccacca gggcaagggt ctgggcagcg
     1741 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     1801 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     1861 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgaacgc
     1921 gttaagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     1981 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     2041 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     2101 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     2161 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     2221 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     2281 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     2341 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     2401 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     2461 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     2521 gtcgacttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     2581 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     2641 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     2701 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     2761 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     2821 tagcagtacg tatagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     2881 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag
     2941 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3001 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg
     3061 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3121 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3181 ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg
     3241 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     3301 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     3361 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     3421 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3481 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3541 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3601 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     3661 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3721 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     3781 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3841 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3901 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3961 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4021 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4081 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4141 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4201 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4261 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     4321 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4381 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     4441 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     4501 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     4561 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     4621 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     4681 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     4741 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     4801 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     4861 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4921 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4981 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5041 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5101 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5161 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5221 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5281 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5341 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5401 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5461 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5521 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5581 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5641 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5701 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     5761 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     5821 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     5881 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     5941 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     6001 aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa
     6061 gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta
     6121 acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg
     6181 gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt
     6241 ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac
     6301 ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     6361 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     6421 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     6481 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     6541 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     6601 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     6661 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     6721 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     6781 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     6841 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     6901 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     6961 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga
     7021 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     7081 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     7141 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     7201 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     7261 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     7321 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     7381 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     7441 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
     7501 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
     7561 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
     7621 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
     7681 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
     7741 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccag agagggccta
     7801 tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga gataattaga
     7861 attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa
     7921 tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc
     7981 gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc
     8041 gtagtcactt ctgatgatgg ggttttagag ctagaaatag caagttaaaa taaggctagt
     8101 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttaagcttg gcgtaactag
     8161 atcttgagac aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
     8221 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     8281 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt aca
//
