LOCUS       sequence-495733-        8333 bp    DNA     circular SYN 18-DEC-2025
DEFINITION  A CRISPR-based technology to study chromosome-specific aneuploidy by
            targeting human centromeres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8333)
  AUTHORS   Bosco N, Goldberg A, Zhao X, Mays JC, Cheng P, Johnson AF, Bianchi 
            JJ, Toscani C, Di Tommaso E, Katsnelson L, Annuar D, Mei S, 
            Faitelson RE, Pesselev IY, Mohamed KS, Mermerian A, 
            Camacho-Hernandez EM, Gionco CA, Manikas J, Tseng YS, Sun Z, Fani S,
            Keegan S, Lippman SM, Fenyo D, Giunta S, Santaguida S, Davoli T
  TITLE     KaryoCreate: A CRISPR-based technology to study chromosome-specific 
            aneuploidy by targeting human centromeres.
  JOURNAL   Cell. 2023 Apr 27;186(9):1985-2001.e19. doi: 
            10.1016/j.cell.2023.03.029. Epub 2023 Apr 18.
  PUBMED    37075754
REFERENCE   2  (bases 1 to 8333)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 18, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2023.03.029"; journalName: "Cell"; date: 
            "2023-04-27- 27"; volume: "186"; issue: "9"; pages: "1985-2001.e19"
COMMENT     Sequence Label: pLenti-gRNA10-1
FEATURES             Location/Qualifiers
     source          1..8333
                     /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_feature    573..690
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        829..2007
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1060..1998
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1955..1975
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2020..2619
                     /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(2020..2039)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2516..2536
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2635..3223
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2688..2708)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3106..3117)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3295..3528
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3606..3727
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3643..3662)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3697..3716
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3767..3902
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3922..3941)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3923..3941)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3951..3968)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3951..3967)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3960..3982)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4109..4564
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4196..4215)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4406..4427
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4590..4694
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4695..4763,4764..5555)
                     /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(4913..4932)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5726..6314
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6215..6234
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6468..6485
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6602..6623
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6638..6643,6644..6661,6662..6668)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6676..6692
                     /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     6681..6703
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6700..6716
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6700..6716
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6735..6755
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6737..6755
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6783..7009
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7010..7190
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7237..7362
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7855..8088
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8273..8317
                     /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 gtttgaggac
      421 ttcgttggaa ggtttaagag ctatgctgga aacagcatag caagtttaaa taaggctagt
      481 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttaagcttg gcgtaactag
      541 atcttgagac aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
      601 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
      661 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
      721 ctttggcgcc ggctcgaggg ggcccgggtg caaagatgga taaagtttta aacagagagg
      781 aatctttgca gctaatggac cttctaggtc ttgaaaggag tgggaattgg ctccggtgcc
      841 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
      901 aattgatccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
      961 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     1021 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc
     1081 cgcgggcctg gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacctgg
     1141 ctgcagtacg tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg
     1201 ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg
     1261 ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct
     1321 ctagccattt aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt
     1381 gtaaatgcgg gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac
     1441 ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg
     1501 agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg
     1561 ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg
     1621 gaaagatggc cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg
     1681 ggagagcggg cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc
     1741 gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc
     1801 ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac
     1861 actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa
     1921 tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt
     1981 ttttttcttc catttcaggt gtcgtgacgt acggccacca tgaccgagta caagcccacg
     2041 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     2101 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     2161 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     2221 gacgacggcg ccgccgtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     2281 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     2341 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     2401 gtcggagtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     2461 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     2521 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     2581 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa cgcgttaagt cgacaatcaa
     2641 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     2701 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     2761 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     2821 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     2881 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     2941 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3001 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     3061 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3121 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     3181 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa
     3241 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     3301 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     3361 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     3421 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     3481 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt acgtatagta
     3541 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     3601 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3661 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3721 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac
     3781 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3841 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3901 cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc
     3961 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     4021 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     4081 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     4141 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4201 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4261 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4321 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4381 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4441 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4501 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     4561 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4621 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4681 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4741 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4801 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4861 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4921 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4981 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5041 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5101 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5161 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5221 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5281 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5341 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5401 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5461 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5521 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5581 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5641 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5701 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5761 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5821 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5881 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5941 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6001 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6061 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6121 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6181 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6241 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6301 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6361 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6421 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     6481 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     6541 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     6601 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     6661 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     6721 attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt
     6781 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
     6841 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
     6901 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
     6961 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
     7021 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     7081 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     7141 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
     7201 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
     7261 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
     7321 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
     7381 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
     7441 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
     7501 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
     7561 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
     7621 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
     7681 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
     7741 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     7801 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     7861 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     7921 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     7981 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     8041 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     8101 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     8161 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     8221 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     8281 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta aca
//
