LOCUS       sequence-380510-        8970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of multi gRNAs targeting hASCL1 promoter to 
            activate human ASCL1 transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8970)
  AUTHORS   Omachi K, Miner JH
  TITLE     Comparative analysis of dCas9-VP64 variants and multiplexed guide 
            RNAs mediating CRISPR activation.
  JOURNAL   PLoS One. 2022 Jun 28;17(6):e0270008. doi: 
            10.1371/journal.pone.0270008. eCollection 2022.
  PUBMED    35763517
REFERENCE   2  (bases 1 to 8970)
  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: "PLoS One.";
            date: "2022-06-28"; volume: "17(6):e0270008. doi"; pages: " 
            10.1371/journal.pone.0270008. eCollection 2022"
COMMENT     Sequence Label: LentiU6-hASCL1 gRNA_multi1-3-MS2-Puro
FEATURES             Location/Qualifiers
     source          1..8970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        180..428
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        466..484
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        536..554
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        597..845
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        883..901
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        953..971
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        1014..1262
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        1299..1317
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1369..1387
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        1466..2644
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1697..2635
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2592..2612
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2657..3256
                     /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(2657..2676)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3153..3173
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    3272..3860
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3325..3345)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3743..3754)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3932..4165
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4243..4364
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4280..4299)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4334..4353
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4404..4539
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4559..4578)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4560..4578)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4588..4605)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4588..4604)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4597..4619)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4746..5201
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4833..4852)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5043..5064
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5227..5331
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5332..5400,5401..6192)
                     /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(5550..5569)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6363..6951
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6852..6871
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7105..7122
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7239..7260
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7275..7280,7281..7298,7299..7305)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7313..7329
                     /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     7318..7340
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7337..7353
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7337..7353
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7372..7392
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7374..7392
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7420..7646
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7647..7827
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7874..7999
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8492..8725
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8910..8954
                     /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"
ORIGIN
        1 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
       61 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
      121 tatcgtttca gacccacctc ccaaccccga ggggacccag aggtaccgct ctagctagcg
      181 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
      241 taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
      301 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata
      361 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac
      421 gaaacaccgc agccgctcgc tgcagcaggt tttagagcta ggccaacatg aggatcaccc
      481 atgtctgcag ggcctagcaa gttaaaataa ggctagtccg ttatcaactt ggccaacatg
      541 aggatcaccc atgtctgcag ggccaagtgg caccgagtcg gtgctttttt ctctaggagg
      601 gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt agagagataa
      661 ttagaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt
      721 aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc
      781 ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa
      841 acaccgtgga gagtttgcaa ggagcgtttt agagctaggc caacatgagg atcacccatg
      901 tctgcagggc ctagcaagtt aaaataaggc tagtccgtta tcaacttggc caacatgagg
      961 atcacccatg tctgcagggc caagtggcac cgagtcggtg ctttttttct agagagggcc
     1021 tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga gagataatta
     1081 gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat
     1141 aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta
     1201 ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca
     1261 ccgtttattc agccgggagt cgttttagag ctaggccaac atgaggatca cccatgtctg
     1321 cagggcctag caagttaaaa taaggctagt ccgttatcaa cttggccaac atgaggatca
     1381 cccatgtctg cagggccaag tggcaccgag tcggtgcttt tttctagaag aattcgctag
     1441 ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt cagtgggcag agcgcacatc
     1501 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgatccggtg cctagagaag
     1561 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     1621 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     1681 tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg
     1741 ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga ttcttgatcc
     1801 cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc
     1861 gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg
     1921 gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg
     1981 acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca
     2041 cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac
     2101 atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca
     2161 agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc
     2221 ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc
     2281 tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc
     2341 cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta
     2401 ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg
     2461 ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt
     2521 taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc
     2581 ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc
     2641 gtgacgtacg gccaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     2701 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     2761 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     2821 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg ccgtggcggt
     2881 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     2941 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     3001 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggagtctcgc ccgaccacca
     3061 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     3121 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     3181 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     3241 caagcccggt gcctgaacgc gttaagtcga caatcaacct ctggattaca aaatttgtga
     3301 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     3361 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     3421 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     3481 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     3541 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     3601 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     3661 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     3721 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3781 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3841 cctttgggcc gcctccccgc gtcgacttta agaccaatga cttacaaggc agctgtagat
     3901 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     3961 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     4021 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     4081 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     4141 tagtcagtgt ggaaaatctc tagcagtacg tatagtagtt catgtcatct tattattcag
     4201 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     4261 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4321 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     4381 ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4441 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     4501 agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc
     4561 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     4621 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     4681 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     4741 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4801 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4861 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4921 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4981 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5041 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5101 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5161 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     5221 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5281 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5341 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5401 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5461 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5521 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5581 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5641 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5701 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5761 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5821 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5881 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5941 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6001 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6061 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6121 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6181 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6241 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6301 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6361 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6421 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6481 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6541 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6601 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6661 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6721 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6781 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6841 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6901 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6961 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7021 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7081 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     7141 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     7201 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     7261 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     7321 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
     7381 cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct
     7441 tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc
     7501 accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac
     7561 agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat
     7621 ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg
     7681 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     7741 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     7801 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
     7861 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
     7921 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
     7981 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
     8041 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     8101 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     8161 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     8221 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     8281 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     8341 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     8401 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     8461 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     8521 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     8581 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     8641 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     8701 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     8761 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     8821 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     8881 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     8941 aatgggcaag tttgtggaat tggtttaaca
//
