LOCUS       sequence-377685-       10290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry and contains a cassette for U6 driven sgRNA 
            transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10290)
  AUTHORS   Thomsen EA, Skipper KA, Andersen S, Haslund D, Skov TW, Mikkelsen JG
  TITLE     CRISPR-Cas9-directed gene tagging using a single integrase-defective
            lentiviral vector carrying a transposase-based Cas9 off switch.
  JOURNAL   Mol Ther Nucleic Acids. 2022 Aug 4;29:563-576. doi: 
            10.1016/j.omtn.2022.08.005. eCollection 2022 Sep 13.
  PUBMED    36090759
REFERENCE   2  (bases 1 to 10290)
  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: "Mol Ther 
            Nucleic Acids."; date: "2022-08-4"; volume: "29:563-576. doi"; 
            pages: " 10.1016/j.omtn.2022.08.005. eCollection 2022 Sep 13"
COMMENT     Sequence Label: pLV/Guide-mCherry
FEATURES             Location/Qualifiers
     source          1..10290
                     /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"
     CDS             1272..1985
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYKN"
     primer_bind     complement(1417..1435)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1685..1704
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2000..2588
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2053..2073)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2471..2482)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2660..2893
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2971..3092
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3008..3027)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3062..3081
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3132..3267
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3287..3306)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3288..3306)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3316..3333)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3316..3332)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3325..3347)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3474..3929
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3561..3580)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3771..3792
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3955..4059
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4060..4128,4129..4920)
                     /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(4278..4297)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5091..5679
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5580..5599
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5833..5850
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5967..5988
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6003..6008,6009..6026,6027..6033)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6041..6057
                     /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     6046..6068
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6065..6081
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6065..6081
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6100..6120
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6102..6120
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6148..6374
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6375..6555
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6602..6727
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7220..7453
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7638..7682
                     /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        7860..8108
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7860..7880
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8031..8050
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9990..10065
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    10121..10238
                     /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 gatggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
     1321 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
     1381 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
     1441 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
     1501 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
     1561 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
     1621 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     1681 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     1741 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     1801 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     1861 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     1921 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaaga
     1981 attaacgcgt taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     2041 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     2101 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     2161 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     2221 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     2281 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2341 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     2401 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     2461 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     2521 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     2581 ctccccgcgt cgactttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     2641 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
     2701 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     2761 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     2821 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     2881 aaaatctcta gcagtacgta tagtagttca tgtcatctta ttattcagta tttataactt
     2941 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     3001 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3061 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     3121 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3181 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3241 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
     3301 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     3361 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     3421 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     3481 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3541 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3601 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3661 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3721 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3781 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3841 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3901 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     3961 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4021 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4081 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4141 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4201 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4261 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4321 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4381 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4441 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4501 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4561 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4621 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4681 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4741 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4801 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4861 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4921 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4981 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5041 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5101 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5161 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5221 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5281 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5341 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5401 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5461 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5521 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5581 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5641 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5701 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5761 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     5821 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     5881 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     5941 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     6001 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     6061 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
     6121 gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca
     6181 acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc
     6241 gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga
     6301 catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag
     6361 ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct
     6421 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     6481 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     6541 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     6601 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     6661 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     6721 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     6781 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     6841 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     6901 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     6961 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     7021 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca
     7081 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
     7141 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
     7201 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
     7261 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
     7321 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
     7381 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
     7441 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
     7501 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
     7561 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
     7621 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
     7681 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
     7741 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
     7801 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccagag
     7861 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
     7921 taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
     7981 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata
     8041 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac
     8101 gaaacaccgg agacggttgt aaatgagcac acaaaataca catgctaaaa tattatattc
     8161 tatgaccttt ataaaatcaa ccaaaatctt ctttttaata actttagtat caataattag
     8221 aatttttatg ttcctttttg caaactttta ataaaaatga gcaaaataaa aaaacgctag
     8281 ttttagtaac tcgcgttgtt ttcttcacct ttaataatag ctactccacc acttgttcct
     8341 aagcggtcag ctcctgcttc aatcattttt tgagcatctt caaatgttct aactccacca
     8401 gctgctttaa ctaaagcatt gtctttaaca actgacttca ttagtttaac atcttcaaat
     8461 gttgcacctg attttgaaaa tcctgttgat gttttaacaa attctaatcc agcttcaaca
     8521 gctatttcac aagctttcat gatttcttct tttgttaata aacaattttc cataatacat
     8581 ttaacaacat gtgatccagc tgcttttttt acagctttca tgtcttctaa aactaattca
     8641 taatttttgt cttttaatgc accaatattt aataccatat caatttctgt tgcaccatct
     8701 ttaattgctt cagaaacttc gaatgctttt gtagctgttg tgcatgcacc tagaggaaaa
     8761 cctacaacat ttgttattcc tacatttgtg ccttttaata attctttaca atagcttgtt
     8821 caatatgaat taacacaaac tgttgcaaaa tcaaattcaa ttgcttcatc acataattgt
     8881 ttaatttcag ctttcgtagc atcttgtttt aataatgtgt gatctatata tttgtttagt
     8941 ttcatttttt ctcctatata ttcattttta attttaattc tttaataatt tcgtctactt
     9001 taactttagc gttttgaaca gattcaccaa cacctataaa ataaattttt agtttaggtt
     9061 cagttccact tgggcgaaca gcaaatcatg acttatcttc taaataaaat tttagtaagt
     9121 cttgtcctgg catattatac attccatcga tgtagtcttc aacattaaca actttaagtc
     9181 cagcaatttg agttaagggt gttgctctca atgatttcat taatggttca atttttaatt
     9241 tcttttcttc tggtttaaaa ttcaagttta aagtgaaagt gtaatatgca cccatttctt
     9301 taaataaatc ttctaaatag tctactaatg ttttattttg ttttttataa aatcaagcag
     9361 cctctgctat taatatagaa gcttgtattc catctttatc tctagctgag tcatcaatta
     9421 catatccata actttcttca taagcaaaaa caaaatttaa tccgttatct tcttctttag
     9481 caatttctct acccattcat ttaaatccag ttaaagtttt tacaatatta actccatatt
     9541 tttcatgagc gattctatca cccaaatcac ttgttacaaa acttgaatat agagccggat
     9601 tttttggaat gctatttaag cgttttagat ttgataattt tcaatcaatt aaaattggtc
     9661 ctgtttgatt tccatctaat cttacaaaat gaccatcatg ttttattgcc attccaaatc
     9721 tgtcagcatc tgggtcattc ataataataa tatctgcatc atgtttaata ccatattcaa
     9781 gcggtatttt tcatgcagga tcaaattctg gatttggatt tacaacattt ttaaatgttt
     9841 catcttcaaa tgcatgctct tcaacctcaa taacgttata tcctgattca cgtaatattt
     9901 ttggggtaaa tttagttcct gttccattaa ctgcgctaaa aataattttt aaatcttttt
     9961 tagcttcttg ctcttttttg tacgtctctg ttttagagct agaaatagca agttaaaata
    10021 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt taagcttggc
    10081 gtaactagat cttgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
    10141 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
    10201 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
    10261 gagatccact ttggcgccgg ctcgaggggg
//
