LOCUS       sequence-379903-        8387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of sgRNA targeting hASCL1 promoter to activate
            human ASCL1 transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8387)
  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 8387)
  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_1-MS2-Puro
FEATURES             Location/Qualifiers
     source          1..8387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        162..410
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     162..182
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     333..352
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        448..466
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        518..536
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_feature    627..744
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        883..2061
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1114..2052
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2009..2029
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2074..2673
                     /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(2074..2093)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2570..2590
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2689..3277
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2742..2762)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3160..3171)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3349..3582
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3660..3781
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3697..3716)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3751..3770
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3821..3956
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3976..3995)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3977..3995)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4005..4022)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4005..4021)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4014..4036)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4163..4618
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4250..4269)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4460..4481
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4644..4748
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4749..4817,4818..5609)
                     /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(4967..4986)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5780..6368
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6269..6288
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6522..6539
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6656..6677
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6692..6697,6698..6715,6716..6722)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6730..6746
                     /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     6735..6757
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6754..6770
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6754..6770
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6789..6809
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6791..6809
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6837..7063
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7064..7244
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7291..7416
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7909..8142
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8327..8371
                     /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 agagggccta tttcccatga
      181 ttccttcata tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga
      241 ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt
      301 agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa
      361 agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc gcagccgctc
      421 gctgcagcag gttttagagc taggccaaca tgaggatcac ccatgtctgc agggcctagc
      481 aagttaaaat aaggctagtc cgttatcaac ttggccaaca tgaggatcac ccatgtctgc
      541 agggccaagt ggcaccgagt cggtgctttt tttggatccg cttggcgtaa ctagatcttg
      601 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
      661 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
      721 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccactttgg
      781 cgccggctcg agggggcccg ggtgcaaaga tggataaagt tttaaacaga gaggaatctt
      841 tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag
      901 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
      961 tccggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     1021 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     1081 ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg
     1141 cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac ctggctgcag
     1201 tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc gaggccttgc
     1261 gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc
     1321 cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc
     1381 atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag tcttgtaaat
     1441 gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc
     1501 cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc accgagaatc
     1561 ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc gccgccgtgt
     1621 atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg agcggaaaga
     1681 tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag
     1741 cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca
     1801 tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc gagcttttgg
     1861 agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc ccacactgag
     1921 tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt ggaatttgcc
     1981 ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca aagttttttt
     2041 cttccatttc aggtgtcgtg acgtacggcc accatgaccg agtacaagcc cacggtgcgc
     2101 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     2161 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     2221 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     2281 ggcgccgccg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     2341 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     2401 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcgga
     2461 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     2521 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     2581 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     2641 acctggtgca tgacccgcaa gcccggtgcc tgaacgcgtt aagtcgacaa tcaacctctg
     2701 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     2761 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     2821 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     2881 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     2941 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     3001 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     3061 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     3121 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     3181 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     3241 cagacgagtc ggatctccct ttgggccgcc tccccgcgtc gactttaaga ccaatgactt
     3301 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     3361 ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     3421 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     3481 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     3541 gatccctcag acccttttag tcagtgtgga aaatctctag cagtacgtat agtagttcat
     3601 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     3661 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3721 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3781 atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc
     3841 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3901 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3961 gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     4021 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     4081 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     4141 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4201 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4261 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4321 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4381 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4441 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4501 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4561 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     4621 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4681 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4741 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4801 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4861 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4921 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4981 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5041 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5101 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5161 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5221 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5281 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5341 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5401 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5461 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5521 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5581 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5641 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5701 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5761 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5821 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5881 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5941 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6001 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6061 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6121 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6181 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6241 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6301 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6361 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6421 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     6481 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     6541 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     6601 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     6661 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     6721 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     6781 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa gcttaatgta
     6841 gtcttatgca atactcttgt agtcttgcaa catggtaacg atgagttagc aacatgcctt
     6901 acaaggagag aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc
     6961 cttattagga aggcaacaga cgggtctgac atggattgga cgaaccactg aattgccgca
     7021 ttgcagagat attgtattta agtgcctagc tcgatacata aacgggtctc tctggttaga
     7081 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     7141 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     7201 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
     7261 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
     7321 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
     7381 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
     7441 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
     7501 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
     7561 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
     7621 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
     7681 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc
     7741 accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg
     7801 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac
     7861 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt
     7921 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
     7981 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
     8041 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
     8101 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
     8161 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
     8221 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     8281 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     8341 attattggaa ttagataaat gggcaagttt gtggaattgg tttaaca
//
