LOCUS       sequence-136378-       10260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses S. pyogenes CRISPR chimeric RNA element with customizable 
            sgRNA from U6 promoter and Crimson reporter from EF-1a promoter. 
            Lentiviral backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10260)
  AUTHORS   Canver MC, Smith EC, Sher F, Pinello L, Sanjana NE, Shalem O, Chen 
            DD, Schupp PG, Vinjamur DS, Garcia SP, Luc S, Kurita R, Nakamura Y, 
            Fujiwara Y, Maeda T, Yuan GC, Zhang F, Orkin SH, Bauer DE
  TITLE     BCL11A enhancer dissection by Cas9-mediated in situ saturating 
            mutagenesis.
  JOURNAL   Nature. 2015 Sep 16. doi: 10.1038/nature15521.
  PUBMED    26375006
REFERENCE   2  (bases 1 to 10260)
  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: "Nature. 
            2015 Sep 16. doi: 10.1038/nature15521."
COMMENT     Sequence Label: lentiGuide-Crimson
FEATURES             Location/Qualifiers
     source          1..10260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        87..1264
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          318..1255
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1212..1232
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1271..1280
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1277..1954
                     /codon_start=1
                     /product="far-red noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2009)"
                     /label=E2-Crimson
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGVGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFFYGSKAYIKHPADIPDYLKQSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTLIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERNYPRDGVLKGENHM
                     ALKLKGGGHYLCEFKSIYMAKKPVKLPGYHYVDYKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     1675..1694
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1970..2558
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2023..2043)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2441..2452)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2630..2863
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2941..3062
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2978..2997)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3032..3051
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3102..3237
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3257..3276)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3258..3276)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3286..3303)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3286..3302)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3295..3317)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3444..3899
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3531..3550)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3741..3762
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3925..4029
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4030..4098,4099..4890)
                     /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(4248..4267)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5061..5649
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5550..5569
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5803..5820
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5937..5958
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5973..5978,5979..5996,5997..6003)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6011..6027
                     /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     6016..6038
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6035..6051
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6035..6051
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6070..6090
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6072..6090
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6118..6344
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6345..6525
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6572..6697
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7190..7423
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7608..7652
                     /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        7830..8078
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7830..7850
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8001..8020
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9960..10035
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    10091..10208
                     /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 ccactggctg cagtacgtga ttcttgatcc
      421 cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc
      481 gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg
      541 gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg
      601 acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca
      661 cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac
      721 atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca
      781 agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc
      841 ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc
      901 tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc
      961 cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta
     1021 ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg
     1081 ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt
     1141 taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc
     1201 ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc
     1261 gtgacgtacg gccaccatgg atagcactga gaacgtcatc aagcccttca tgcgcttcaa
     1321 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg tgggcgaggg
     1381 caagccctac gagggcaccc agaccgccaa gctgcaagtg accaagggcg gccccctgcc
     1441 cttcgcctgg gacatcctgt ccccccagtt cttctacggc tccaaggcgt acatcaagca
     1501 ccccgccgac atccccgact acctcaagca gtccttcccc gagggcttca agtgggagcg
     1561 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     1621 cggcaccctc atctaccacg tgaagttcat cggcgtgaac ttcccctccg acggccccgt
     1681 aatgcagaag aagactctgg gctgggagcc ctccactgag cgcaactacc cccgcgacgg
     1741 cgtgctgaag ggcgagaacc acatggcgct gaagctgaag ggcggcggcc actacctgtg
     1801 tgagttcaag tccatctaca tggccaagaa gcccgtgaag ctgcccggct accactacgt
     1861 ggactacaag ctcgacatca cctcccacaa cgaggactac accgtggtgg agcagtacga
     1921 gcgcgccgag gcccgccacc acctgttcca gtagacgcgt taagtcgaca atcaacctct
     1981 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2041 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2101 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2161 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2221 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2281 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2341 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     2401 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     2461 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     2521 tcagacgagt cggatctccc tttgggccgc ctccccgcgt cgactttaag accaatgact
     2581 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     2641 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     2701 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     2761 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     2821 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtacgta tagtagttca
     2881 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     2941 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3001 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3061 tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc
     3121 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3181 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3241 ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     3301 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     3361 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     3421 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3481 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3541 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3601 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3661 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3721 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3781 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3841 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     3901 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3961 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4021 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4081 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4141 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4201 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4261 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4321 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4381 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4441 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4501 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4561 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4621 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4681 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4741 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4801 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4861 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4921 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4981 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5041 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5101 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5161 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5221 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5281 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5341 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5401 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5461 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5521 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5581 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5641 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5701 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     5761 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     5821 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     5881 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     5941 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     6001 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     6061 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt
     6121 agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct
     6181 tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg
     6241 ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc
     6301 attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag
     6361 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     6421 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     6481 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag
     6541 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga
     6601 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
     6661 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
     6721 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
     6781 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
     6841 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
     6901 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
     6961 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
     7021 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt
     7081 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
     7141 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
     7201 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg
     7261 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
     7321 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
     7381 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
     7441 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
     7501 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca
     7561 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
     7621 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc
     7681 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt
     7741 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga
     7801 cccacctccc aaccccgagg ggacccagag agggcctatt tcccatgatt ccttcatatt
     7861 tgcatatacg atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa
     7921 agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt
     7981 taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt
     8041 tcttggcttt atatatcttg tggaaaggac gaaacaccgg agacggttgt aaatgagcac
     8101 acaaaataca catgctaaaa tattatattc tatgaccttt ataaaatcaa ccaaaatctt
     8161 ctttttaata actttagtat caataattag aatttttatg ttcctttttg caaactttta
     8221 ataaaaatga gcaaaataaa aaaacgctag ttttagtaac tcgcgttgtt ttcttcacct
     8281 ttaataatag ctactccacc acttgttcct aagcggtcag ctcctgcttc aatcattttt
     8341 tgagcatctt caaatgttct aactccacca gctgctttaa ctaaagcatt gtctttaaca
     8401 actgacttca ttagtttaac atcttcaaat gttgcacctg attttgaaaa tcctgttgat
     8461 gttttaacaa attctaatcc agcttcaaca gctatttcac aagctttcat gatttcttct
     8521 tttgttaata aacaattttc cataatacat ttaacaacat gtgatccagc tgcttttttt
     8581 acagctttca tgtcttctaa aactaattca taatttttgt cttttaatgc accaatattt
     8641 aataccatat caatttctgt tgcaccatct ttaattgctt cagaaacttc gaatgctttt
     8701 gtagctgttg tgcatgcacc tagaggaaaa cctacaacat ttgttattcc tacatttgtg
     8761 ccttttaata attctttaca atagcttgtt caatatgaat taacacaaac tgttgcaaaa
     8821 tcaaattcaa ttgcttcatc acataattgt ttaatttcag ctttcgtagc atcttgtttt
     8881 aataatgtgt gatctatata tttgtttagt ttcatttttt ctcctatata ttcattttta
     8941 attttaattc tttaataatt tcgtctactt taactttagc gttttgaaca gattcaccaa
     9001 cacctataaa ataaattttt agtttaggtt cagttccact tgggcgaaca gcaaatcatg
     9061 acttatcttc taaataaaat tttagtaagt cttgtcctgg catattatac attccatcga
     9121 tgtagtcttc aacattaaca actttaagtc cagcaatttg agttaagggt gttgctctca
     9181 atgatttcat taatggttca atttttaatt tcttttcttc tggtttaaaa ttcaagttta
     9241 aagtgaaagt gtaatatgca cccatttctt taaataaatc ttctaaatag tctactaatg
     9301 ttttattttg ttttttataa aatcaagcag cctctgctat taatatagaa gcttgtattc
     9361 catctttatc tctagctgag tcatcaatta catatccata actttcttca taagcaaaaa
     9421 caaaatttaa tccgttatct tcttctttag caatttctct acccattcat ttaaatccag
     9481 ttaaagtttt tacaatatta actccatatt tttcatgagc gattctatca cccaaatcac
     9541 ttgttacaaa acttgaatat agagccggat tttttggaat gctatttaag cgttttagat
     9601 ttgataattt tcaatcaatt aaaattggtc ctgtttgatt tccatctaat cttacaaaat
     9661 gaccatcatg ttttattgcc attccaaatc tgtcagcatc tgggtcattc ataataataa
     9721 tatctgcatc atgtttaata ccatattcaa gcggtatttt tcatgcagga tcaaattctg
     9781 gatttggatt tacaacattt ttaaatgttt catcttcaaa tgcatgctct tcaacctcaa
     9841 taacgttata tcctgattca cgtaatattt ttggggtaaa tttagttcct gttccattaa
     9901 ctgcgctaaa aataattttt aaatcttttt tagcttcttg ctcttttttg tacgtctctg
     9961 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
    10021 gcaccgagtc ggtgcttttt taagcttggc gtaactagat cttgagacaa atggcagtat
    10081 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
    10141 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
    10201 aaaattttcg ggtttattac agggacagca gagatccact ttggcgccgg ctcgaggggg
//
