LOCUS       sequence-436108-        8459 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing mClover3 along with U6 driven hybrid 
            guide (hgRNA) targeting two intergenic sites in the human genome 
            using SpCas9 and AsCas12a nucleases (CHyMErA system), respectively.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8459)
  AUTHORS   Xiao MS, Damodaran AP, Kumari B, Dickson E, Xing K, On TA, Parab N, 
            King HE, Perez AR, Guiblet WM, Duncan G, Che A, Chari R, Andresson 
            T, Vidigal JA, Weatheritt RJ, Aregger M, Gonatopoulos-Pournatzis T
  TITLE     Genome-scale exon perturbation screens uncover exons critical for 
            cell fitness.
  JOURNAL   Mol Cell. 2024 Jul 11;84(13):2553-2572.e19. doi: 
            10.1016/j.molcel.2024.05.024. Epub 2024 Jun 24.
  PUBMED    38917794
REFERENCE   2  (bases 1 to 8459)
  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: 
            "10.1016/j.molcel.2024.05.024"; journalName: "Mol Cell"; date: 
            "2024-07-11- 11"; volume: "84"; issue: "13"; pages: "2553-2572.e19"
COMMENT     Sequence Label: pLCHKOv3-mClover-intergenic_3-As
FEATURES             Location/Qualifiers
     source          1..8459
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             48..644
                     /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(48..67)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     544..564
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             660..713
                     /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             join(720..722,723..1436)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(762..783)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1370..1391
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1451..2039
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1504..1524)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1922..1933)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2111..2344
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2422..2543
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2459..2478)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2513..2532
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2583..2718
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2738..2757)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2739..2757)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2767..2784)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2767..2783)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2776..2798)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2925..3380
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3012..3031)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3222..3243
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3406..3510
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3511..3579,3580..4371)
                     /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(3729..3748)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4542..5130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5031..5050
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5284..5301
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5418..5439
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5454..5459,5460..5477,5478..5484)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5492..5508
                     /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     5497..5519
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5516..5532
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5516..5532
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5551..5571
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5553..5571
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5599..5825
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5826..6006
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6053..6178
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6671..6904
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7089..7133
                     /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        complement(7499..7747)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(7557..7576)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(7727..7747)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    7789..7906
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        join(7975..8459,1..26)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     8404..8422
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
ORIGIN
        1 accgaatcac cgacctctct ccccaggggg atccaccgga gcttaccatg accgagtaca
       61 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
      121 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
      181 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
      241 gggtcgcgga cgacggcgcc gccgtggcgg tctggaccac gccggagagc gtcgaagcgg
      301 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
      361 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
      421 tggccaccgt cggtgtttcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
      481 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
      541 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
      601 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgccacgcgt ggcagtggag
      661 agggcagagg aagtctgcta acatgcggtg acgtcgagga gaatcctggc ccactcgagg
      721 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
      781 acgtaaacgg ccacaagttc agcgtccgcg gcgagggcga gggcgatgcc accaacggca
      841 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
      901 tgaccacctt cggctacggc gtggcctgct tcagccgcta ccccgaccac atgaagcagc
      961 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atctctttca
     1021 aggacgacgg tacctacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1081 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1141 tggagtacaa cttcaacagc cactacgtct atatcacggc cgacaagcag aagaactgca
     1201 tcaaggctaa cttcaagatc cgccacaacg ttgaggacgg cagcgtgcag ctcgccgacc
     1261 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1321 tgagccatca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1381 tggagttcgt gaccgccgcc gggattacac atggcatgga cgagctgtac aagtaacgcg
     1441 ttaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     1501 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     1561 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     1621 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     1681 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     1741 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     1801 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     1861 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     1921 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     1981 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg
     2041 tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     2101 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     2161 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     2221 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     2281 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     2341 agcagtacgt atagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa
     2401 tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc
     2461 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2521 tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc
     2581 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2641 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     2701 gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc
     2761 gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     2821 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     2881 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     2941 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3001 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3061 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3121 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3181 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3241 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3301 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     3361 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3421 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3481 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3541 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3601 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3661 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3721 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3781 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3841 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3901 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3961 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4021 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4081 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4141 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4201 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4261 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4321 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4381 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4441 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4501 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4561 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4621 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4681 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4741 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4801 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4861 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4921 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4981 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5041 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5101 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5161 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5221 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5281 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     5341 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     5401 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     5461 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     5521 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag
     5581 ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa
     5641 cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg
     5701 aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg
     5761 gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca
     5821 taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     5881 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     5941 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     6001 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
     6061 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
     6121 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
     6181 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
     6241 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
     6301 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
     6361 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
     6421 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
     6481 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag
     6541 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
     6601 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
     6661 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
     6721 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
     6781 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     6841 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     6901 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     6961 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     7021 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     7081 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     7141 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     7201 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     7261 attcaccatt atcgtttcag acccacctcc caaccccgag gggaccctgc aggatttaaa
     7321 tccgagggga cccagagaat tcaaaaaaac acaacaggat aacatacttg tatctacaag
     7381 agtagaaatt agcaccgact cggtgccact ttttcaagtt gataacggac tagccttatt
     7441 tcaacttgct atgctgtttc cagcatagct ctgaaacgcc tccatcttca gtcagtacgg
     7501 tacctcgtcc tttccacaag atatataaag ccaagaaatc gaaatacttt caagttacgg
     7561 taagcatatg atagtccatt ttaaaacata attttaaaac tgcaaactac ccaagaaatt
     7621 attactttct acgtcacgta ttttgtacta atatctttgt gtttacagtc aaattaattc
     7681 taattatctc tctaacagcc ttgtatcgta tatgcaaata tgaaggaatc atgggaaata
     7741 ggccctcgaa ttctagatct tgagacaaat ggcagtattc atccacaatt ttaaaagaaa
     7801 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     7861 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     7921 ggacagcaga gatccacttt ggcgccggct cgagatttaa atcctgcagg cgagggggcc
     7981 cgggttgcgc cttttccaag gcagccctgg gtttgcgcag ggacgcggct gctctgggcg
     8041 tggttccggg aaacgcagcg gcgccgaccc tgggtctcgc acattcttca cgtccgttcg
     8101 cagcgtcacc cggatcttcg ccgctaccct tgtgggcccc ccggcgacgc ttcctgctcc
     8161 gcccctaagt cgggaaggtt ccttgcggtt cgcggcgtgc cggacgtgac aaacggaagc
     8221 cgcagctcac actagtaccc tcgcagacgg acagcgccag ggagcaatgg cagcgcgccg
     8281 accgcgatgg gctgtggcca atagcggctg ctcagcaggg cgcgccgaga gcagcggccg
     8341 ggaaggggcg gtgcgggagg cggggtgtgg ggcggtagtg tgggccctgt tcctgcccgc
     8401 gcggtgttcc gcattctgca agcctccgga gcgcacgtcg gcagtcggct ccctcgttg
//
