LOCUS       sequence-495732-        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-gRNA4-3
FEATURES             Location/Qualifiers
     source          1..8333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(81..92)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             270..503
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    581..702
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(618..637)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     672..691
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      742..877
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(897..916)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(898..916)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(926..943)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(926..942)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(935..957)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1084..1539
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1171..1190)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1381..1402
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1565..1669
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1670..1738,1739..2530)
                     /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(1888..1907)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2701..3289
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3190..3209
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3443..3460
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3577..3598
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3613..3618,3619..3636,3637..3643)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3651..3667
                     /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     3656..3678
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3675..3691
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3675..3691
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3710..3730
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3712..3730
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3758..3984
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             3985..4165
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4212..4337
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4830..5063
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5248..5292
                     /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        5470..5718
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5470..5490
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5641..5660
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    5881..5998
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6137..7315
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6368..7306
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7263..7283
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             7328..7927
                     /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(7328..7347)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7824..7844
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    join(7943..8333,1..198)
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7996..8016)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
       61 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
      121 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
      181 tttgggccgc ctccccgcgt cgactttaag accaatgact tacaaggcag ctgtagatct
      241 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
      301 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
      361 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
      421 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
      481 gtcagtgtgg aaaatctcta gcagtacgta tagtagttca tgtcatctta ttattcagta
      541 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
      601 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
      661 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
      721 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
      781 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
      841 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct
      901 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
      961 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     1021 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     1081 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     1141 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     1201 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     1261 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     1321 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     1381 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     1441 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     1501 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     1561 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1621 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1681 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1741 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1801 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1861 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     1921 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1981 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     2041 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     2101 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     2161 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     2221 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     2281 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2341 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2401 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2461 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2521 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2581 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2641 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2701 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2761 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2821 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     2881 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2941 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3001 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3061 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3121 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3181 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3241 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3301 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3361 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3421 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     3481 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     3541 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     3601 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     3661 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     3721 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg
     3781 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac
     3841 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag
     3901 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt
     3961 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag
     4021 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     4081 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     4141 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     4201 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     4261 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     4321 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     4381 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     4441 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     4501 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     4561 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     4621 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     4681 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     4741 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     4801 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     4861 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     4921 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     4981 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     5041 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     5101 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     5161 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     5221 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     5281 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     5341 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     5401 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     5461 ggacccagag agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct
     5521 gttagagaga taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg
     5581 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg
     5641 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg
     5701 tggaaaggac gaaacaccgc cttttgtaga atctgcaggg tttaagagct atgctggaaa
     5761 cagcatagca agtttaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
     5821 ggtgcttttt taagcttggc gtaactagat cttgagacaa atggcagtat tcatccacaa
     5881 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
     5941 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
     6001 ggtttattac agggacagca gagatccact ttggcgccgg ctcgaggggg cccgggtgca
     6061 aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct tctaggtctt
     6121 gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     6181 ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa ggtggcgcgg
     6241 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     6301 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     6361 aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc
     6421 ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc ccgagcttcg
     6481 ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc
     6541 ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg
     6601 cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc
     6661 gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat
     6721 ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc
     6781 gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg
     6841 gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct
     6901 ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg
     6961 gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag
     7021 gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc
     7081 gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga
     7141 ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc
     7201 ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat
     7261 tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgacgtac
     7321 ggccaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     7381 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     7441 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     7501 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gccgtggcgg tctggaccac
     7561 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     7621 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     7681 caaggagccc gcgtggttcc tggccaccgt cggagtctcg cccgaccacc agggcaaggg
     7741 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     7801 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     7861 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     7921 tgcctgaacg cgttaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     7981 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     8041 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     8101 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     8161 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     8221 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     8281 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cgg
//
