LOCUS       sequence-457013-       10253 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Backbone for generation of donor vectors for CRISPR/Cas9 mediated 
            Saturation Genome Editing (SGE) of TP53 Exon 7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10253)
  AUTHORS   Funk JS, Klimovich M, Drangenstein D, Pielhoop O, Hunold P, Borowek 
            A, Noeparast M, Pavlakis E, Neumann M, Balourdas DI, Kochhan K, 
            Merle N, Bullwinkel I, Wanzel M, Elmshauser S, Teply-Szymanski J, 
            Nist A, Procida T, Bartkuhn M, Humpert K, Mernberger M, Savai R, 
            Soussi T, Joerger AC, Stiewe T
  TITLE     Deep CRISPR mutagenesis characterizes the functional diversity of 
            TP53 mutations.
  JOURNAL   Nat Genet. 2025 Jan;57(1):140-153. doi: 10.1038/s41588-024-02039-4. 
            Epub 2025 Jan 7.
  PUBMED    39774325
REFERENCE   2  (bases 1 to 10253)
  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"; doi: 
            "10.1038/s41588-024-02039-4"; journalName: "Nat Genet"; date: 
            "2025-01"; volume: "57"; issue: "1"; pages: "140-153"
COMMENT     Sequence Label: HR700_HA2TP53_Ex7
FEATURES             Location/Qualifiers
     source          1..10253
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(805..838)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1115..1326
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1339..1607
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      1635..1644
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1665..2327
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
     CDS             2403..2456
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2457..3056
                     /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(2457..2476)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2953..2973
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3094..3111)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3100..3307
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(3551..3584)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     6440..6459
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      6515..6524
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6521..7651
                     /codon_start=1
                     /product="herpes simplex virus thymidine kinase"
                     /label=HSV TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MASYPGHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRPEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQCGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRSMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     polyA_signal    7652..7773
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7689..7708)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7743..7762
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7807..7823)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7807..7823)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7820..7842)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7831..7847
                     /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)."
     promoter        complement(join(7855..7861,7862..7879,7880..7885))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7900..7921
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8038..8055)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8209..8797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8289..8308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9118..9933)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(9118..9933)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     9841..9860
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9925..9944
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     misc_feature    10052..10070
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     primer_bind     10226..10244
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaaac tagtggtacc tgtacaagag
      421 gctgctcccc ccgtggcccc tgcaccagca gctcctacac cggcggcccc tgcaccagcc
      481 ccctcctggc ccctgtcatc ttctgtccct tcccagaaaa cctaccaggg cagctacggt
      541 ttccgtctgg gcttcttgca ttctgggaca gccaagtctg tgacttgcac ggtcagttgc
      601 cctgaggggc tggcttccat gagacttcaa tgcctggccg tatccccctg catttctttt
      661 gtttggaact ttgggattcc tcttcaccct ttggcttcct gtcagtgttt ttttatagtt
      721 tacccactta atgtgtgatc tctgactcct gtcccaaagt tgaatattcc ccccttgaat
      781 ttgggctttt atccatcggt ctctataact tcgtatagca tacattatac gaagttatgc
      841 tagcgaggga cagccccccc ccaaagcccc cagggatgta attacgtccc tcccccgcta
      901 gggggcagca gcgagccgcc cggggctccg ctccggtccg gcgctccccc cgcatccccg
      961 agccggcagc gtgcggggac agcccgggca cggggaaggt ggcacgggat cgctttcctc
     1021 tgaacgcttc tcgctgctct ttgagcctgc agacacctgg ggggatacgg ggaaaagcta
     1081 gcaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     1141 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg
     1201 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     1261 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     1321 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga
     1381 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga
     1441 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gtccgggcct ttgtccggcg
     1501 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct
     1561 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg
     1621 gcgcctacga attcgccacc atggagagcg acgagagcgg cctgcccgcc atggagatcg
     1681 agtgccgcat caccggcacc ctgaacggcg tggagttcga gctggtgggc ggcggagagg
     1741 gcacccccaa gcagggccgc atgaccaaca agatgaagag caccaaaggc gccctgacct
     1801 tcagccccta cctgctgagc cacgtgatgg gctacggctt ctaccacttc ggcacctacc
     1861 ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa cggcggctac accaacaccc
     1921 gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag cttcagctac cgctacgagg
     1981 ccggccgcgt gatcggcgac ttcaaggtgg tgggcaccgg cttccccgag gacagcgtga
     2041 tcttcaccga caagatcatc cgcagcaacg ccaccgtgga gcacctgcac cccatgggcg
     2101 ataacgtgct ggtgggcagc ttcgcccgca ccttcagcct gcgcgacggc ggctactaca
     2161 gcttcgtggt ggacagccac atgcacttca agagcgccat ccaccccagc atcctgcaga
     2221 acgggggccc catgttcgcc ttccgccgcg tggaggagct gcacagcaac accgagctgg
     2281 gcatcgtgga gtaccagcac gccttcaaga cccccatcgc cttcgccaga tcccgcgctc
     2341 agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg ctccaccgga tctcgcagta
     2401 ctgagggcag aggaagtctc ctaacatgcg gtgacgtgga ggagaatccc ggccctatga
     2461 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
     2521 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
     2581 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     2641 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     2701 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     2761 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     2821 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     2881 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     2941 catccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     3001 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgaggat
     3061 cctgatcgag tctagagggc ccccgctgat cagcctcgac tgtgccttct agttgccagc
     3121 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3181 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3241 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagagaat agcaggcatg
     3301 ctggggaaga tctttttccc cgtatccccc caggtgtctg caggctcaaa gagcagcgag
     3361 aagcgttcag aggaaagcga tcccgtgcca ccttccccgt gcccgggctg tccccgcacg
     3421 ctgccggctc ggggatgcgg ggggagcgcc ggaccggagc ggagccccgg gcggctcgct
     3481 gctgccccct agcgggggag ggacgtaatt acatccctgg gggctttggg ggggggctgt
     3541 ccctagatct ataacttcgt atagcataca ttatacgaag ttatgagaat tcacacgcgt
     3601 ccatcacacc ctcagcatct ctcctgggga tgcagaactt ttctttttct tcatccacgt
     3661 gtattccttg gcttttgaaa ataagctcct gaccaggctt ggtggctcac acctgcaatc
     3721 ccagcactct caaagaggcc aaggcaggca gatcacctga gcccaggagt tcaagaccag
     3781 cctgggtaac atgatgaaac ctcgtctcta caaaaaaata caaaaaatta gccaggcatg
     3841 gtggtgcaca cctatagtcc cagccactta ggaggctgag gtgggaagat cacttgaggc
     3901 caggagatgg aggctgcagt gagctgtgat cacaccactg tgctccagcc tgagtgacag
     3961 agcaagaccc tatctcaaaa aaaaaaaaaa aaaagaaaag ctcctgaggt gtagacgcca
     4021 actctctcta gctcgctagt gggttgcagg aggtgcttac gcatgtttgt ttctttgctg
     4081 ccgaattcca gttgctttat ctgttcactt gtgccctgac tttcaactct gtctccttcc
     4141 tcttcctaca gtactcccct gccctcaaca agatgttttg ccaactggcc aagacctgcc
     4201 ctgtgcagct gtgggttgat tccacacccc cgcccggcac ccgcgtccgc gccatggcca
     4261 tctacaagca gtcacagcac atgacggagg ttgtgaggcg ctgcccccac catgagcgct
     4321 gctcagatag cgatggtgag cagctggggc tggagagacg acagggctgg ttgcccaggg
     4381 tccccaggcc tctgattcct cactgattgc tcttaggtct ggcccctcct cagcatctta
     4441 tccgagtgga aggaaatttg cgtgtggagt atttggatga cagaaacact tttcgacata
     4501 gtgtggtggt gccctatgag ccgcctgagg tctggtttgc aactggggtc tctgggagga
     4561 ggggttaagg gtggttgtca gtggccctcc aggtgagcag taggggggct ttctcctgct
     4621 gcttatttga cctccctata accccatgag atgtgcaaag taaatgggtt taactattgc
     4681 acagttgaaa aaactgaagc ttacagaggc taagggcctc ccctgcttgg ctgggcgcag
     4741 tggctcatgc ctgtaatccc agcactttgg gaggccaagg caggcggatc acgaggttgg
     4801 gagatcgaga ccatcctggc taacggtgaa accccgtctc tactgaaaaa tacaaaaaaa
     4861 aattagccgg gcgtggtgct gggcacctgt agtcccagct actcgggagg ctgaggaagg
     4921 agaatggcgt gaacctgggc ggtggagctt gcagtgagct gagatcacgc cactgcactc
     4981 cagcctgggc gacagagcga gattccatct caaaaaaaaa aaaaaaaggc ctcccctgct
     5041 tgccacaggt ctccccaagg cgcactggcc tcatcttggg gtcttccctg tgttcacttg
     5101 ccgaagacac cctgcacact ggcctgctgt gccccagcct ctgcttgcct ctgacccctg
     5161 ggcccacctc ttaccgattt cttccatact actacccatc cacctctcat cacatccccg
     5221 gcggggaatc tccttactgc tcccactcag ttttcttttc tctggctttg ggacctctta
     5281 acctgtggct tctcctccac ctacctggag ctggagctta ggctccagaa aggacaaggg
     5341 tggttgggag tagatggagc ctggtttttt aaatgggaca ggtaggacct gatttcctta
     5401 ctgcctcttg cttctctttt cctatcctga gtagtggtaa tctactggga cggaacagct
     5461 ttgaggtgcg tgtttgtgcc tgtcctggga gagaccggcg cacagaggaa gagaatctcc
     5521 gcaagaaagg ggagcctcac cacgagctgc ccccagggag cactaagcga ggtaagcaag
     5581 caggacaaga agcggtggag gagaccaagg gtgcagttat gcctcagatt cacttttatc
     5641 acctttcctt gcctctttcc tagcactgcc caacaacacc agctcctctc cccagccaaa
     5701 gaagaaacca ctggatggag aatatttcac ccttcaggta ctaagtcttg ggacctctta
     5761 tcaagtggaa agtttccagt ctaacactca aaatgccgtt ttcttcttga ctgttttacc
     5821 tgcaattggg gcatttgcca tcagggggca gtgatgcctc aaagacaatg gctcctggtt
     5881 gtagctaact aacttcagaa caccaactta taccataata tatattttaa aggaccagac
     5941 cagctttcaa aaagaaaatt gttaaagaga gcatgaaaat ggttctatga ctttgcctga
     6001 tacagatgct acttgactta cgatggtgtt acttcctgat aaactcgtcg taagttgaaa
     6061 atattgtaag ttgaaaatgg atttaataca cctaatctaa ggaacatcat agcttagcct
     6121 agcctgcgtc gacacccctc tggcctcgca cacattccac atccaccggt aggcgccaac
     6181 cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt
     6241 tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca
     6301 ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca
     6361 gcggccaata gcagctttgc tccttcgctt tctgggctca ggggcggggc gggcgcccga
     6421 aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc
     6481 tcctcttcct catctccggg cctttcgacc tgcagccacc atggcttcgt accccggcca
     6541 tcaacacgcc tctgcgttcg accaggctgc gcgttctcgc ggccatagca accgacgtac
     6601 ggcgttgcgc cctcgccggc agcaagaagc cacggaagtc cgcccggagc agaaaatgcc
     6661 cacgctactg cgggtttata tagacggtcc ccacgggatg gggaaaacca ccaccacgca
     6721 actgctggtg gccctgggtt cgcgcgacga tatcgtctac gtacccgagc cgatgactta
     6781 ctggcgggtg ctgggggctt ccgagacaat cgcgaacatc tacaccacac aacaccgcct
     6841 cgaccagggt gagatatcgg ccggggacgc ggcggtggta atgacaagcg cccagataac
     6901 aatgggcatg ccttatgccg tgaccgacgc cgttctggct cctcatatcg ggggggaggc
     6961 tgggagctca catgccccgc ccccggccct caccctcatc ttcgaccgcc atcccatcgc
     7021 cgccctcctg tgctacccgg ccgcgcggta ccttatgggc agcatgaccc cccaggccgt
     7081 gctggcgttc gtggccctca tcccgccgac cttgcccggc accaacatcg tgcttggggc
     7141 ccttccggag gacagacaca tcgaccgcct ggccaaacgc cagcgccccg gcgagcggct
     7201 ggacctggct atgctggctg cgattcgccg cgtttacggg ctacttgcca atacggtgcg
     7261 gtatctgcag tgcggcgggt cgtggcggga ggactgggga cagctttcgg ggacggccgt
     7321 gccgccccag ggtgccgagc cccagagcaa cgcgggccca cgaccccata tcggggacac
     7381 gttatttacc ctgtttcggg cccccgagtt gctggccccc aacggcgacc tgtataacgt
     7441 gtttgcctgg gccttggacg tcttggccaa acgcctccgt tccatgcacg tctttatcct
     7501 ggattacgac caatcgcccg ccggctgccg ggacgccctg ctgcaactta cctccgggat
     7561 ggtccagacc cacgtcacca cccccggctc cataccgacg atatgcgacc tggcgcgcac
     7621 gtttgcccgg gagatggggg aggctaactg aaacttgttt attgcagctt ataatggtta
     7681 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     7741 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatatcaa gcttggcgta
     7801 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     7861 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     7921 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     7981 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     8041 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     8101 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     8161 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     8221 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     8281 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     8341 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     8401 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     8461 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     8521 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     8581 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     8641 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     8701 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     8761 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     8821 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     8881 aaaaaggatc ttcacctaga tccttttgat ccccgccacg gttgatgaga gctttgttgt
     8941 aggtggacca gttggtgatt ttgaactttt gctttgccac ggaacggtct gcgttgtcgg
     9001 gaagatgcgt gatctgatcc ttcaactcag caaaagttcg atttattcaa caaagccgcc
     9061 gtcccgtcaa gtcagcgtaa tgctctgcca gtgttacaac caattaacca attctgatta
     9121 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
     9181 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag
     9241 gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat
     9301 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
     9361 atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc
     9421 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc
     9481 ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
     9541 caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc
     9601 ttctaatacc tggaatgctg tttttccggg gatcgcagtg gtgagtaacc atgcatcatc
     9661 aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag
     9721 tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
     9781 ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt
     9841 atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta atcgcggcct
     9901 cgagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
     9961 agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag
    10021 attttgagac acaattcatc gatgatggtt gagatgtgta taagagacag attattgaag
    10081 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
    10141 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
    10201 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtc
//
