LOCUS       sequence-476817-        9517 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses either functional CRISPR gRNAs from the hU6-promotor and 
            empowers pairing of CRISPR screens with 3' and 5' scRNA-Seq 
            capturing by FACS-sorting on eBFP2+ without prior resistance 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9517)
  AUTHORS   Enk LUB, Hellmig M, Riecken K, Kilian C, Datlinger P, Jauch-Speer 
            SL, Neben T, Sultana Z, Sivayoganathan V, Borchers A, Paust HJ, Zhao
            Y, Asada N, Liu S, Agalioti T, Pelczar P, Wiech T, Bock C, Huber TB,
            Huber S, Bonn S, Gagliani N, Fehse B, Panzer U, Krebs CF
  TITLE     Targeting T cell plasticity in kidney and gut inflammation by pooled
            single-cell CRISPR screening.
  JOURNAL   Sci Immunol. 2024 Jun 14;9(96):eadd6774. doi: 
            10.1126/sciimmunol.add6774. Epub 2024 Jun 14.
  PUBMED    38875317
REFERENCE   2  (bases 1 to 9517)
  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: "Sci 
            Immunol."; date: "2024-06-14"; volume: "9(96):eadd6774. doi"; pages:
            " 10.1126/sciimmunol.add6774. Epub 2024 Jun 14"
COMMENT     Sequence Label: CROPseq-SFFV-eBFP2
FEATURES             Location/Qualifiers
     source          1..9517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      426..1014
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     915..934
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1168..1185
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1302..1323
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1338..1343,1344..1361,1362..1368)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1376..1392
                     /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     1381..1403
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1400..1416
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1400..1416
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1435..1455
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1437..1455
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1483..1709
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1710..1890
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1937..2062
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2555..2788
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2973..3017
                     /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"
     misc_feature    3198..3315
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3410..3816
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      3822..3831
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3828..3830,3831..3833,3834..4547)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(3873..3894)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    4563..5151
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4616..4636)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5034..5045)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        5307..5555
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5307..5327
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5478..5497
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7437..7512
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     LTR             7565..7745
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    7823..7944
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7860..7879)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7914..7933
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7984..8119
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8139..8158)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8140..8158)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8168..8185)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8168..8184)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8177..8199)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8326..8781
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8413..8432)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8623..8644
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8807..8911
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8912..8980,8981..9517,1..255)
                     /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(9130..9149)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
       61 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
      121 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
      181 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
      241 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
      301 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
      361 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
      421 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
      481 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
      541 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
      601 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
      661 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
      721 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
      781 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
      841 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
      901 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
      961 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     1021 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     1081 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     1141 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     1201 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     1261 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     1321 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     1381 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     1441 aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac
     1501 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
     1561 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
     1621 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
     1681 tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct
     1741 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     1801 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     1861 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
     1921 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     1981 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     2041 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     2101 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     2161 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     2221 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     2281 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     2341 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     2401 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     2461 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     2521 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     2581 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     2641 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     2701 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     2761 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     2821 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     2881 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     2941 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     3001 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     3061 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     3121 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagaatctt gagacaaatg
     3181 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     3241 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     3301 aaaattcaaa attttcgggt ttattacagg gacagcagag atccactttg gcgccggctc
     3361 gagggggccg ctgaaagacc ccacctgtag gtttggcaag ctagctgcag taacgccatt
     3421 ttgcaaggca tggaaaaata ccaaaccaag aatagagaag ttcagatcaa gggcgggtac
     3481 atgaaaatag ctaacgttgg gccaaacagg atatctgcgg tgagcagttt cggccccggc
     3541 ccggggccaa gaacagatgg tcaccgcagt ttcggccccg gcccgaggcc aagaacagat
     3601 ggtccccaga tatggcccaa ccctcagcag tttcttaaga cccatcagat gtttccaggc
     3661 tcccccaagg acctgaaatg accctgcgcc ttatttgaat taaccaatca gcctgcttct
     3721 cgcttctgtt cgcgcgcttc tgcttcccga gctctataaa agagctcaca acccctcact
     3781 cggcgcgcca gtcctccgat tgactgagtc gcccggatcc cgccaccatg gtgagcaagg
     3841 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     3901 gccacaagtt cagcgtgagg ggcgagggcg agggcgatgc caccaacggc aagctgaccc
     3961 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     4021 tgagccacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     4081 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     4141 gcacctacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     4201 agctgaaggg cgtcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     4261 acttcaacag ccacaacatc tatatcatgg ccgtcaagca gaagaacggc atcaaggtga
     4321 acttcaagat ccgccacaac gtggaggacg gcagcgtgca gctcgccgac cactaccagc
     4381 agaacacccc catcggcgac ggccccgtgc tgctgcccga cagccactac ctgagcaccc
     4441 agtccgtgct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcc
     4501 gcaccgccgc cgggatcact ctcggcatgg acgagctgta caagtgaacg cgttaagtcg
     4561 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     4621 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     4681 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     4741 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     4801 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     4861 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     4921 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     4981 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     5041 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     5101 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cgtcgacttt
     5161 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
     5221 actggaaggg ctaattcact cccaacgaag acaagatttc accattatcg tttcagaccc
     5281 acctcccaac cccgagggga cccagagagg gcctatttcc catgattcct tcatatttgc
     5341 atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga
     5401 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
     5461 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
     5521 tggctttata tatcttgtgg aaaggacgaa acaccggaga cggttgtaaa tgagcacaca
     5581 aaatacacat gctaaaatat tatattctat gacctttata aaatcaacca aaatcttctt
     5641 tttaataact ttagtatcaa taattagaat ttttatgttc ctttttgcaa acttttaata
     5701 aaaatgagca aaataaaaaa acgctagttt tagtaactcg cgttgttttc ttcaccttta
     5761 ataatagcta ctccaccact tgttcctaag cggtcagctc ctgcttcaat cattttttga
     5821 gcatcttcaa atgttctaac tccaccagct gctttaacta aagcattgtc tttaacaact
     5881 gacttcatta gtttaacatc ttcaaatgtt gcacctgatt ttgaaaatcc tgttgatgtt
     5941 ttaacaaatt ctaatccagc ttcaacagct atttcacaag ctttcatgat ttcttctttt
     6001 gttaataaac aattttccat aatacattta acaacatgtg atccagctgc tttttttaca
     6061 gctttcatgt cttctaaaac taattcataa tttttgtctt ttaatgcacc aatatttaat
     6121 accatatcaa tttctgttgc accatcttta attgcttcag aaacttcgaa tgcttttgta
     6181 gctgttgtgc atgcacctag aggaaaacct acaacatttg ttattcctac atttgtgcct
     6241 tttaataatt ctttacaata gcttgttcaa tatgaattaa cacaaactgt tgcaaaatca
     6301 aattcaattg cttcatcaca taattgttta atttcagctt tcgtagcatc ttgttttaat
     6361 aatgtgtgat ctatatattt gtttagtttc attttttctc ctatatattc atttttaatt
     6421 ttaattcttt aataatttcg tctactttaa ctttagcgtt ttgaacagat tcaccaacac
     6481 ctataaaata aatttttagt ttaggttcag ttccacttgg gcgaacagca aatcatgact
     6541 tatcttctaa ataaaatttt agtaagtctt gtcctggcat attatacatt ccatcgatgt
     6601 agtcttcaac attaacaact ttaagtccag caatttgagt taagggtgtt gctctcaatg
     6661 atttcattaa tggttcaatt tttaatttct tttcttctgg tttaaaattc aagtttaaag
     6721 tgaaagtgta atatgcaccc atttctttaa ataaatcttc taaatagtct actaatgttt
     6781 tattttgttt tttataaaat caagcagcct ctgctattaa tatagaagct tgtattccat
     6841 ctttatctct agctgagtca tcaattacat atccataact ttcttcataa gcaaaaacaa
     6901 aatttaatcc gttatcttct tctttagcaa tttctctacc cattcattta aatccagtta
     6961 aagtttttac aatattaact ccatattttt catgagcgat tctatcaccc aaatcacttg
     7021 ttacaaaact tgaatataga gccggatttt ttggaatgct atttaagcgt tttagatttg
     7081 ataattttca atcaattaaa attggtcctg tttgatttcc atctaatctt acaaaatgac
     7141 catcatgttt tattgccatt ccaaatctgt cagcatctgg gtcattcata ataataatat
     7201 ctgcatcatg tttaatacca tattcaagcg gtatttttca tgcaggatca aattctggat
     7261 ttggatttac aacattttta aatgtttcat cttcaaatgc atgctcttca acctcaataa
     7321 cgttatatcc tgattcacgt aatatttttg gggtaaattt agttcctgtt ccattaactg
     7381 cgctaaaaat aatttttaaa tcttttttag cttcttgctc ttttttgtac gtctctgttt
     7441 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
     7501 ccgagtcggt gcttttttaa gcttggcgta actagatctt gagacactgc tttttgcttg
     7561 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     7621 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     7681 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     7741 tagcagtacg tatagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     7801 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag
     7861 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     7921 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg
     7981 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     8041 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     8101 ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg
     8161 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     8221 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     8281 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     8341 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     8401 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     8461 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     8521 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     8581 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     8641 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     8701 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     8761 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     8821 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     8881 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     8941 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     9001 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     9061 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     9121 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     9181 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     9241 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     9301 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     9361 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     9421 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     9481 actattaact ggcgaactac ttactctagc ttcccgg
//
