LOCUS       sequence-255092-       10261 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses S. pyogenes CRISPR chimeric RNA element with customizable 
            sgRNA from U6 promoter and DsREd from EF-1a promoter. 3rd generation
            lentiviral backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10261)
  TITLE     Emory Custom Cloning Core Plasmids - Oskar Laur
REFERENCE   2  (bases 1 to 10261)
  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"
COMMENT     Sequence Label: plenti guide ds red
FEATURES             Location/Qualifiers
     source          1..10261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        87..1265
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          318..1256
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1213..1233
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1272..1281
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1278..1955
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(1458..1478)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1676..1695
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1870..1893
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    1971..2559
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2024..2044)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2442..2453)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2631..2864
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2942..3063
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2979..2998)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3033..3052
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3103..3238
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3258..3277)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3259..3277)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3287..3304)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3287..3303)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3296..3318)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3445..3900
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3532..3551)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3742..3763
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3926..4030
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4031..4099,4100..4891)
                     /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(4249..4268)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5062..5650
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5551..5570
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5804..5821
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5938..5959
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5974..5979,5980..5997,5998..6004)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6012..6028
                     /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     6017..6039
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6036..6052
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6036..6052
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6071..6091
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6073..6091
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6119..6345
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6346..6526
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6573..6698
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7191..7424
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7609..7653
                     /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        7831..8079
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7831..7851
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8002..8021
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9961..10036
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    10092..10209
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cccgggtgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
       61 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
      121 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa
      181 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
      241 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
      301 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
      361 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
      421 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
      481 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
      541 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
      601 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
      661 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
      721 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
      781 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
      841 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
      901 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
      961 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     1021 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     1081 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     1141 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     1201 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     1261 cgtgacgtac ggccaccatg gatagcactg agaacgtcat caagcccttc atgcgcttca
     1321 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     1381 gcaagcccta cgagggcacc cagaccgcca agctgcaggt gaccaagggc ggccccctgc
     1441 ccttcgcctg ggacatcctg tccccccagt tccagtacgg ctccaaggtg tacgtgaagc
     1501 accccgccga catccccgac tacaagaagc tgtccttccc cgagggcttc aagtgggagc
     1561 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     1621 acggcacctt catctaccac gtgaagttca tcggcgtgaa cttcccctcc gacggccccg
     1681 taatgcagaa gaagactctg ggctgggagc cctccaccga gcgcctgtac ccccgcgacg
     1741 gcgtgctgaa gggcgagatc cacaaggcgc tgaagctgaa gggcggcggc cactacctgg
     1801 tggagttcaa gtcaatctac atggccaaga agcccgtgaa gctgcccggc tactactacg
     1861 tggactccaa gctggacatc acctcccaca acgaggacta caccgtggtg gagcagtacg
     1921 agcgcgccga ggcccgccac cacctgttcc agtagacgcg ttaagtcgac aatcaacctc
     1981 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     2041 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     2101 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     2161 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     2221 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     2281 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     2341 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     2401 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     2461 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     2521 ctcagacgag tcggatctcc ctttgggccg cctccccgcg tcgactttaa gaccaatgac
     2581 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     2641 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
     2701 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     2761 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     2821 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtacgt atagtagttc
     2881 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     2941 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3001 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3061 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg
     3121 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3181 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3241 gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     3301 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     3361 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     3421 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3481 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3541 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3601 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3661 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3721 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3781 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3841 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     3901 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3961 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4021 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4081 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4141 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4201 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4261 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4321 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4381 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4441 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4501 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4561 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4621 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4681 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4741 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4801 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4861 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4921 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4981 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5041 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5101 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5161 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5221 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5281 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5341 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5401 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5461 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5521 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5581 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5641 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5701 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     5761 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     5821 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     5881 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     5941 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     6001 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     6061 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg
     6121 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
     6181 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
     6241 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
     6301 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
     6361 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     6421 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     6481 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     6541 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     6601 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     6661 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     6721 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     6781 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     6841 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     6901 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     6961 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     7021 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     7081 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     7141 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     7201 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     7261 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     7321 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     7381 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     7441 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     7501 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     7561 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     7621 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     7681 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     7741 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     7801 acccacctcc caaccccgag gggacccaga gagggcctat ttcccatgat tccttcatat
     7861 ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca
     7921 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     7981 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
     8041 ttcttggctt tatatatctt gtggaaagga cgaaacaccg gagacggttg taaatgagca
     8101 cacaaaatac acatgctaaa atattatatt ctatgacctt tataaaatca accaaaatct
     8161 tctttttaat aactttagta tcaataatta gaatttttat gttccttttt gcaaactttt
     8221 aataaaaatg agcaaaataa aaaaacgcta gttttagtaa ctcgcgttgt tttcttcacc
     8281 tttaataata gctactccac cacttgttcc taagcggtca gctcctgctt caatcatttt
     8341 ttgagcatct tcaaatgttc taactccacc agctgcttta actaaagcat tgtctttaac
     8401 aactgacttc attagtttaa catcttcaaa tgttgcacct gattttgaaa atcctgttga
     8461 tgttttaaca aattctaatc cagcttcaac agctatttca caagctttca tgatttcttc
     8521 ttttgttaat aaacaatttt ccataataca tttaacaaca tgtgatccag ctgctttttt
     8581 tacagctttc atgtcttcta aaactaattc ataatttttg tcttttaatg caccaatatt
     8641 taataccata tcaatttctg ttgcaccatc tttaattgct tcagaaactt cgaatgcttt
     8701 tgtagctgtt gtgcatgcac ctagaggaaa acctacaaca tttgttattc ctacatttgt
     8761 gccttttaat aattctttac aatagcttgt tcaatatgaa ttaacacaaa ctgttgcaaa
     8821 atcaaattca attgcttcat cacataattg tttaatttca gctttcgtag catcttgttt
     8881 taataatgtg tgatctatat atttgtttag tttcattttt tctcctatat attcattttt
     8941 aattttaatt ctttaataat ttcgtctact ttaactttag cgttttgaac agattcacca
     9001 acacctataa aataaatttt tagtttaggt tcagttccac ttgggcgaac agcaaatcat
     9061 gacttatctt ctaaataaaa ttttagtaag tcttgtcctg gcatattata cattccatcg
     9121 atgtagtctt caacattaac aactttaagt ccagcaattt gagttaaggg tgttgctctc
     9181 aatgatttca ttaatggttc aatttttaat ttcttttctt ctggtttaaa attcaagttt
     9241 aaagtgaaag tgtaatatgc acccatttct ttaaataaat cttctaaata gtctactaat
     9301 gttttatttt gttttttata aaatcaagca gcctctgcta ttaatataga agcttgtatt
     9361 ccatctttat ctctagctga gtcatcaatt acatatccat aactttcttc ataagcaaaa
     9421 acaaaattta atccgttatc ttcttcttta gcaatttctc tacccattca tttaaatcca
     9481 gttaaagttt ttacaatatt aactccatat ttttcatgag cgattctatc acccaaatca
     9541 cttgttacaa aacttgaata tagagccgga ttttttggaa tgctatttaa gcgttttaga
     9601 tttgataatt ttcaatcaat taaaattggt cctgtttgat ttccatctaa tcttacaaaa
     9661 tgaccatcat gttttattgc cattccaaat ctgtcagcat ctgggtcatt cataataata
     9721 atatctgcat catgtttaat accatattca agcggtattt ttcatgcagg atcaaattct
     9781 ggatttggat ttacaacatt tttaaatgtt tcatcttcaa atgcatgctc ttcaacctca
     9841 ataacgttat atcctgattc acgtaatatt tttggggtaa atttagttcc tgttccatta
     9901 actgcgctaa aaataatttt taaatctttt ttagcttctt gctctttttt gtacgtctct
     9961 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
    10021 ggcaccgagt cggtgctttt ttaagcttgg cgtaactaga tcttgagaca aatggcagta
    10081 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
    10141 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
    10201 caaaattttc gggtttatta cagggacagc agagatccac tttggcgccg gctcgagggg
    10261 g
//
