LOCUS       sequence-437895-        9549 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 9549)
  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 9549)
  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-MPSV-eBFP2
FEATURES             Location/Qualifiers
     source          1..9549
                     /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"
     regulatory      3854..3863
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3860..3862,3863..3865,3866..4579)
                     /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(3905..3926)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    4595..5183
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4648..4668)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5066..5077)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        5339..5587
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5339..5359
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5510..5529
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7469..7544
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     LTR             7597..7777
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    7855..7976
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7892..7911)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7946..7965
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8016..8151
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8171..8190)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8172..8190)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8200..8217)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8200..8216)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8209..8231)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8358..8813
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8445..8464)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8655..8676
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8839..8943
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8944..9012,9013..9549,1..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(9162..9181)
                     /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 ctagcttaag taacgccatt
     3421 ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca
     3481 gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     3541 cagggccaag aacagttgga acagcagaat atgggccaaa caggatatct gtggtaagca
     3601 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc
     3661 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     3721 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     3781 gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg attgactgag
     3841 tcgcccggat cccgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     3901 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtga ggggcgaggg
     3961 cgagggcgat gccaccaacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     4021 gcccgtgccc tggcccaccc tcgtgaccac cctgagccac ggcgtgcagt gcttcgcccg
     4081 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     4141 ccaggagcgc accatcttct tcaaggacga cggcacctac aagacccgcg ccgaggtgaa
     4201 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcgtcgact tcaaggagga
     4261 cggcaacatc ctggggcaca agctggagta caacttcaac agccacaaca tctatatcat
     4321 ggccgtcaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acgtggagga
     4381 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     4441 gctgctgccc gacagccact acctgagcac ccagtccgtg ctgagcaaag accccaacga
     4501 gaagcgcgat cacatggtcc tgctggagtt ccgcaccgcc gccgggatca ctctcggcat
     4561 ggacgagctg tacaagtgaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg
     4621 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     4681 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     4741 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     4801 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4861 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     4921 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     4981 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     5041 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     5101 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     5161 ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta
     5221 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     5281 agacaagatt tcaccattat cgtttcagac ccacctccca accccgaggg gacccagaga
     5341 gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat
     5401 aattagaatt aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa
     5461 gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat
     5521 gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg
     5581 aaacaccgga gacggttgta aatgagcaca caaaatacac atgctaaaat attatattct
     5641 atgaccttta taaaatcaac caaaatcttc tttttaataa ctttagtatc aataattaga
     5701 atttttatgt tcctttttgc aaacttttaa taaaaatgag caaaataaaa aaacgctagt
     5761 tttagtaact cgcgttgttt tcttcacctt taataatagc tactccacca cttgttccta
     5821 agcggtcagc tcctgcttca atcatttttt gagcatcttc aaatgttcta actccaccag
     5881 ctgctttaac taaagcattg tctttaacaa ctgacttcat tagtttaaca tcttcaaatg
     5941 ttgcacctga ttttgaaaat cctgttgatg ttttaacaaa ttctaatcca gcttcaacag
     6001 ctatttcaca agctttcatg atttcttctt ttgttaataa acaattttcc ataatacatt
     6061 taacaacatg tgatccagct gcttttttta cagctttcat gtcttctaaa actaattcat
     6121 aatttttgtc ttttaatgca ccaatattta ataccatatc aatttctgtt gcaccatctt
     6181 taattgcttc agaaacttcg aatgcttttg tagctgttgt gcatgcacct agaggaaaac
     6241 ctacaacatt tgttattcct acatttgtgc cttttaataa ttctttacaa tagcttgttc
     6301 aatatgaatt aacacaaact gttgcaaaat caaattcaat tgcttcatca cataattgtt
     6361 taatttcagc tttcgtagca tcttgtttta ataatgtgtg atctatatat ttgtttagtt
     6421 tcattttttc tcctatatat tcatttttaa ttttaattct ttaataattt cgtctacttt
     6481 aactttagcg ttttgaacag attcaccaac acctataaaa taaattttta gtttaggttc
     6541 agttccactt gggcgaacag caaatcatga cttatcttct aaataaaatt ttagtaagtc
     6601 ttgtcctggc atattataca ttccatcgat gtagtcttca acattaacaa ctttaagtcc
     6661 agcaatttga gttaagggtg ttgctctcaa tgatttcatt aatggttcaa tttttaattt
     6721 cttttcttct ggtttaaaat tcaagtttaa agtgaaagtg taatatgcac ccatttcttt
     6781 aaataaatct tctaaatagt ctactaatgt tttattttgt tttttataaa atcaagcagc
     6841 ctctgctatt aatatagaag cttgtattcc atctttatct ctagctgagt catcaattac
     6901 atatccataa ctttcttcat aagcaaaaac aaaatttaat ccgttatctt cttctttagc
     6961 aatttctcta cccattcatt taaatccagt taaagttttt acaatattaa ctccatattt
     7021 ttcatgagcg attctatcac ccaaatcact tgttacaaaa cttgaatata gagccggatt
     7081 ttttggaatg ctatttaagc gttttagatt tgataatttt caatcaatta aaattggtcc
     7141 tgtttgattt ccatctaatc ttacaaaatg accatcatgt tttattgcca ttccaaatct
     7201 gtcagcatct gggtcattca taataataat atctgcatca tgtttaatac catattcaag
     7261 cggtattttt catgcaggat caaattctgg atttggattt acaacatttt taaatgtttc
     7321 atcttcaaat gcatgctctt caacctcaat aacgttatat cctgattcac gtaatatttt
     7381 tggggtaaat ttagttcctg ttccattaac tgcgctaaaa ataattttta aatctttttt
     7441 agcttcttgc tcttttttgt acgtctctgt tttagagcta gaaatagcaa gttaaaataa
     7501 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt aagcttggcg
     7561 taactagatc ttgagacact gctttttgct tgtactgggt ctctctggtt agaccagatc
     7621 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     7681 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     7741 ctcagaccct tttagtcagt gtggaaaatc tctagcagta cgtatagtag ttcatgtcat
     7801 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     7861 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     7921 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     7981 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     8041 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     8101 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
     8161 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     8221 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     8281 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     8341 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     8401 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     8461 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     8521 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     8581 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     8641 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     8701 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     8761 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     8821 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     8881 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     8941 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     9001 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     9061 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     9121 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     9181 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     9241 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     9301 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     9361 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     9421 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     9481 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     9541 gcttcccgg
//
