LOCUS       Exported               10183 bp ds-DNA     circular SYN 12-MAY-2021
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-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10183)
  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 10183)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10183
                     /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"
     CDS             3870..4469
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3870..3889)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4366..4386
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4485..5073
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4538..4558)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4956..4967)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5145..5378
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5456..5577
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5493..5512)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5547..5566
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5617..5752
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5679..5698
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(5772..5791)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5773..5791)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5801..5818)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5801..5817)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5810..5832)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5959..6414
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6046..6065)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6256..6277
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6440..6544
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6545..7405
                     /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(6763..6782)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7576..8164
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8065..8084
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8318..8335
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8452..8473
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8488..8518
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8526..8542
                     /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     8531..8553
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8550..8566
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8550..8566
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8585..8605
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8587..8605
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8633..8859
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8860..9040
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9087..9212
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9705..9938
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10123..10167
                     /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 tgaccgagta caagcccacg gtgcgcctcg
     3901 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     3961 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     4021 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     4081 ccgccgtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     4141 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     4201 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggagtct
     4261 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     4321 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     4381 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     4441 ggtgcatgac ccgcaagccc ggtgcctgaa cgcgttaagt cgacaatcaa cctctggatt
     4501 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     4561 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     4621 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     4681 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     4741 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     4801 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     4861 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     4921 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     4981 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     5041 cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa
     5101 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     5161 ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
     5221 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     5281 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     5341 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt acgtatagta gttcatgtca
     5401 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt
     5461 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5521 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5581 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     5641 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     5701 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg
     5761 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
     5821 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     5881 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     5941 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     6001 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     6061 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     6121 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     6181 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     6241 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     6301 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     6361 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     6421 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     6481 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     6541 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     6601 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     6661 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     6721 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     6781 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     6841 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6901 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6961 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     7021 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     7081 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     7141 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     7201 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     7261 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     7321 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     7381 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     7441 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     7501 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     7561 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     7621 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     7681 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     7741 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     7801 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     7861 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     7921 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     7981 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     8041 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     8101 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     8161 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     8221 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     8281 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     8341 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     8401 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     8461 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     8521 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     8581 gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct
     8641 tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa
     8701 ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta
     8761 ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc
     8821 agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag
     8881 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     8941 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     9001 tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact
     9061 tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc
     9121 gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag
     9181 gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc
     9241 gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag
     9301 tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag
     9361 aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac
     9421 ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa
     9481 aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg
     9541 cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga
     9601 agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag
     9661 gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt
     9721 gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag
     9781 gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag
     9841 gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc
     9901 ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga
     9961 aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa
    10021 cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc
    10081 ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta
    10141 ttggaattag ataaatgggc aagtttgtgg aattggttta aca
//
