LOCUS       Exported               10287 bp ds-DNA     circular SYN 03-JUN-2022
DEFINITION  Expresses S. pyogenes CRISPR chimeric RNA element with customizable 
            sgRNA from U6 promoter and puromycin resistance from EF-1a promoter.
            3rd generation lentiviral backbone.
ACCESSION   .
VERSION     .
KEYWORDS    lentiGuide-dsRed
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10287)
  AUTHORS   Sanjana NE, Shalem O, Zhang F
  TITLE     Improved vectors and genome-wide libraries for CRISPR screening.
  JOURNAL   Nat Methods. 2014 Aug;11(8):783-4. doi: 10.1038/nmeth.3047.
  PUBMED    25075903
REFERENCE   2  (bases 1 to 10287)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Feb 20, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10287
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        162..402
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     162..182
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     333..352
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2292..2367
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2423..2540
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2679..3857
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2910..3848
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3805..3825
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     3840..3899
                     /label=dsRED FP
     regulatory      3864..3873
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3870..4559
                     /codon_start=1
                     /label=DsRED
                     /translation="MATTMASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKG
                     EIHKALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAE
                     GRHHLFL"
     CDS             3882..4559
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(4544..4603)
                     /label=dsRED RP
     misc_feature    4589..5177
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4642..4662)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5060..5071)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5249..5482
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5560..5681
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5597..5616)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5651..5670
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5721..5856
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5783..5802
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(5806..5817)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(5876..5895)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5877..5895)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5905..5922)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5905..5921)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5914..5936)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6063..6518
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6150..6169)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6360..6381
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6544..6648
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6649..7509
                     /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(6867..6886)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7680..8268
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8169..8188
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8422..8439
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8556..8577
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8592..8622
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8630..8646
                     /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     8635..8657
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8654..8670
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8654..8670
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8689..8709
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8691..8709
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8737..8963
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8964..9144
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9191..9316
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9809..10042
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10227..10271
                     /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"
ORIGIN
        1 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
       61 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
      121 tatcgtttca gacccacctc ccaaccccga ggggacccag agagggccta tttcccatga
      181 ttccttcata tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga
      241 ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt
      301 agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa
      361 agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc ggagacggtt
      421 gtaaatgagc acacaaaata cacatgctaa aatattatat tctatgacct ttataaaatc
      481 aaccaaaatc ttctttttaa taactttagt atcaataatt agaattttta tgttcctttt
      541 tgcaaacttt taataaaaat gagcaaaata aaaaaacgct agttttagta actcgcgttg
      601 ttttcttcac ctttaataat agctactcca ccacttgttc ctaagcggtc agctcctgct
      661 tcaatcattt tttgagcatc ttcaaatgtt ctaactccac cagctgcttt aactaaagca
      721 ttgtctttaa caactgactt cattagttta acatcttcaa atgttgcacc tgattttgaa
      781 aatcctgttg atgttttaac aaattctaat ccagcttcaa cagctatttc acaagctttc
      841 atgatttctt cttttgttaa taaacaattt tccataatac atttaacaac atgtgatcca
      901 gctgcttttt ttacagcttt catgtcttct aaaactaatt cataattttt gtcttttaat
      961 gcaccaatat ttaataccat atcaatttct gttgcaccat ctttaattgc ttcagaaact
     1021 tcgaatgctt ttgtagctgt tgtgcatgca cctagaggaa aacctacaac atttgttatt
     1081 cctacatttg tgccttttaa taattcttta caatagcttg ttcaatatga attaacacaa
     1141 actgttgcaa aatcaaattc aattgcttca tcacataatt gtttaatttc agctttcgta
     1201 gcatcttgtt ttaataatgt gtgatctata tatttgttta gtttcatttt ttctcctata
     1261 tattcatttt taattttaat tctttaataa tttcgtctac tttaacttta gcgttttgaa
     1321 cagattcacc aacacctata aaataaattt ttagtttagg ttcagttcca cttgggcgaa
     1381 cagcaaatca tgacttatct tctaaataaa attttagtaa gtcttgtcct ggcatattat
     1441 acattccatc gatgtagtct tcaacattaa caactttaag tccagcaatt tgagttaagg
     1501 gtgttgctct caatgatttc attaatggtt caatttttaa tttcttttct tctggtttaa
     1561 aattcaagtt taaagtgaaa gtgtaatatg cacccatttc tttaaataaa tcttctaaat
     1621 agtctactaa tgttttattt tgttttttat aaaatcaagc agcctctgct attaatatag
     1681 aagcttgtat tccatcttta tctctagctg agtcatcaat tacatatcca taactttctt
     1741 cataagcaaa aacaaaattt aatccgttat cttcttcttt agcaatttct ctacccattc
     1801 atttaaatcc agttaaagtt tttacaatat taactccata tttttcatga gcgattctat
     1861 cacccaaatc acttgttaca aaacttgaat atagagccgg attttttgga atgctattta
     1921 agcgttttag atttgataat tttcaatcaa ttaaaattgg tcctgtttga tttccatcta
     1981 atcttacaaa atgaccatca tgttttattg ccattccaaa tctgtcagca tctgggtcat
     2041 tcataataat aatatctgca tcatgtttaa taccatattc aagcggtatt tttcatgcag
     2101 gatcaaattc tggatttgga tttacaacat ttttaaatgt ttcatcttca aatgcatgct
     2161 cttcaacctc aataacgtta tatcctgatt cacgtaatat ttttggggta aatttagttc
     2221 ctgttccatt aactgcgcta aaaataattt ttaaatcttt tttagcttct tgctcttttt
     2281 tgtacgtctc tgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
     2341 cttgaaaaag tggcaccgag tcggtgcttt tttaagcttg gcgtaactag atcttgagac
     2401 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     2461 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     2521 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca ctttggcgcc
     2581 ggctcgaggg ggcccgggtg caaagatgga taaagtttta aacagagagg aatctttgca
     2641 gctaatggac cttctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg
     2701 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg
     2761 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     2821 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     2881 ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg
     2941 gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg
     3001 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt
     3061 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg
     3121 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt
     3181 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg
     3241 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg
     3301 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac
     3361 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg
     3421 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc
     3481 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg
     3541 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg
     3601 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta
     3661 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg
     3721 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt
     3781 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc
     3841 catttcaggt gtcgtgacgt acggccacca tggccacaac catggcctcc tccgaggacg
     3901 tcatcaagga gttcatgcgc ttcaaggtgc gcatggaggg ctccgtgaac ggccacgagt
     3961 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     4021 aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc cagttccagt
     4081 acggctccaa ggtgtacgtg aagcaccccg ccgacatccc cgactacaag aagctgtcct
     4141 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     4201 tgacccagga ctcctccctg caggacggct ccttcatcta caaggtgaag ttcatcggcg
     4261 tgaacttccc ctccgacggc cccgtaatgc agaagaagac tatgggctgg gaggcctcca
     4321 ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga gatccacaag gccctgaagc
     4381 tgaaggacgg cggccactac ctggtggagt tcaagtctat ctatatggcc aagaagcccg
     4441 tgcagctgcc cggctactac tacgtggact ccaagctgga catcacctcc cacaacgagg
     4501 actacaccat cgtggagcag tacgagcgcg ccgagggccg ccaccacctg ttcctgtagt
     4561 gaattaatta tgaacgcgtt aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     4621 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     4681 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     4741 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     4801 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     4861 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     4921 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     4981 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     5041 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     5101 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     5161 ttgggccgcc tccccgcgtc gactttaaga ccaatgactt acaaggcagc tgtagatctt
     5221 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
     5281 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     5341 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     5401 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     5461 tcagtgtgga aaatctctag cagtacgtat agtagttcat gtcatcttat tattcagtat
     5521 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat
     5581 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5641 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta
     5701 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5761 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     5821 tattccagaa gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta
     5881 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     5941 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     6001 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     6061 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     6121 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     6181 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     6241 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     6301 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     6361 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     6421 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     6481 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     6541 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     6601 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     6661 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     6721 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     6781 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     6841 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     6901 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     6961 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     7021 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     7081 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     7141 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     7201 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     7261 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     7321 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     7381 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     7441 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     7501 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     7561 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     7621 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     7681 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     7741 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     7801 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     7861 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     7921 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     7981 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     8041 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     8101 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     8161 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     8221 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     8281 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     8341 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     8401 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     8461 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     8521 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     8581 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     8641 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct
     8701 cactaaaggg aacaaaagct ggagctgcaa gcttaatgta gtcttatgca atactcttgt
     8761 agtcttgcaa catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc
     8821 gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga
     8881 cgggtctgac atggattgga cgaaccactg aattgccgca ttgcagagat attgtattta
     8941 agtgcctagc tcgatacata aacgggtctc tctggttaga ccagatctga gcctgggagc
     9001 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     9061 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     9121 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa
     9181 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga
     9241 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga
     9301 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg
     9361 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga
     9421 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat
     9481 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa
     9541 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc
     9601 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc
     9661 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
     9721 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
     9781 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
     9841 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
     9901 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
     9961 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
    10021 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
    10081 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
    10141 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
    10201 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
    10261 gggcaagttt gtggaattgg tttaaca
//
