LOCUS       Exported               13500 bp ds-DNA     circular SYN 29-JUL-2021
DEFINITION  lenti sgRNA cloning backbone with MS2 loops at tetraloop and 
            stemloop 2, dCas9-VP64, and blast resistance marker. Contains BsmBI 
            sites for insertion of spacer sequences..
ACCESSION   .
VERSION     .
KEYWORDS    lentiSAMv2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13500)
  AUTHORS   Joung J, Konermann S, Gootenberg JS, Abudayyeh OO, Platt RJ, Brigham
            MD, Sanjana NE, Zhang F
  TITLE     Genome-scale CRISPR-Cas9 knockout and transcriptional activation 
            screening.
  JOURNAL   Nat Protoc. 2017 Apr;12(4):828-863. doi: 10.1038/nprot.2017.016. 
            Epub 2017 Mar 23.
  PUBMED    28333914
REFERENCE   2  (bases 1 to 13500)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 29, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..13500
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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"
     misc_feature    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2607..2847
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     2607..2627
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2778..2797
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2893..2911
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        2963..2981
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        3068..3279
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             3296..3343
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             3344..7444
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7499..7519
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7544..7693
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             7715..7768
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             7769..8167
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8192..8780
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8245..8265)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8663..8674)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8783..8799)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8784..8800)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9305..9485
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9511..9528)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9517..9741
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9787..10215
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9874..9893)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10084..10105
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10224..10244)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10229..10558
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10409..10544
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10471..10490
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10606..10653
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10672..11046
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    11176..11297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11213..11232)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11267..11286
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11346..11362)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11346..11362)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11359..11381)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11370..11386
                     /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)."
     promoter        complement(11394..11424)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11439..11460
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11577..11594)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11748..12336)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11828..11847)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(12507..13367)
                     /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     13130..13149
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13368..13472)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta gctagcgagg gcctatttcc catgattcct tcatatttgc
     2641 atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga
     2701 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
     2761 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
     2821 tggctttata tatcttgtgg aaaggacgaa acaccggaga cgggataccg tctctgtttt
     2881 agagctaggc caacatgagg atcacccatg tctgcagggc ctagcaagtt aaaataaggc
     2941 tagtccgtta tcaacttggc caacatgagg atcacccatg tctgcagggc caagtggcac
     3001 cgagtcggtg ctttttttgg atcctaggtc ttgaaaggag tgggaattgg ctccggtgcc
     3061 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     3121 aattgatccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     3181 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     3241 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg cgtacggcca ccatgaaaag
     3301 gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aaggacaaga agtacagcat
     3361 cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt
     3421 gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct
     3481 gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac
     3541 cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag
     3601 caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt
     3661 ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc
     3721 ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga
     3781 caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca
     3841 cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca
     3901 gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga
     3961 cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc
     4021 ccagctgccc ggcgagaaga agaatggcct gttcggcaac ctgattgccc tgagcctggg
     4081 cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag
     4141 caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc
     4201 cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag
     4261 agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga
     4321 gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta
     4381 caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc
     4441 cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga
     4501 ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa
     4561 cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga
     4621 agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg
     4681 catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag
     4741 aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc
     4801 cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt
     4861 gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt
     4921 gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc
     4981 catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga
     5041 ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt
     5101 caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct
     5161 ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga
     5221 ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt
     5281 gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat
     5341 caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg
     5401 cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga
     5461 catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct
     5521 ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct
     5581 cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa
     5641 ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg
     5701 catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca
     5761 gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga
     5821 actggacatc aaccggctgt ccgactacga tgtggaccac atcgtgcctc agagctttct
     5881 gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaaggccc ggggcaagag
     5941 cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct
     6001 gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg
     6061 cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat
     6121 cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga
     6181 caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg
     6241 gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc
     6301 ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga
     6361 gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca
     6421 ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa
     6481 gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg
     6541 cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct
     6601 gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa
     6661 agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga
     6721 ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc
     6781 caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac
     6841 catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta
     6901 caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga
     6961 aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc
     7021 cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg
     7081 ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga
     7141 cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct
     7201 ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga
     7261 gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt
     7321 tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct
     7381 gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg
     7441 cgacagcgct ggaggaggtg gaagcggagg aggaggaagc ggaggaggag gtagcggacc
     7501 taagaaaaag aggaaggtgg cggccgctgg atccggacgg gctgacgcat tggacgattt
     7561 tgatctggat atgctgggaa gtgacgccct cgatgatttt gaccttgaca tgcttggttc
     7621 ggatgccctt gatgactttg acctcgacat gctcggcagt gacgcccttg atgatttcga
     7681 cctggacatg ctgattaact gtacaggcag tggagagggc agaggaagtc tgctaacatg
     7741 cggtgacgtc gaggagaatc ctggcccaat ggccaagcct ttgtctcaag aagaatccac
     7801 cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt
     7861 cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt
     7921 tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg
     7981 caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg
     8041 acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag
     8101 tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga
     8161 gggctaagaa ttcgatatca agcttatcgg taatcaacct ctggattaca aaatttgtga
     8221 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     8281 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     8341 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     8401 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     8461 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     8521 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     8581 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     8641 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     8701 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     8761 cctttgggcc gcctccccgc atcgataccg tcgacctcga gacctagaaa aacatggagc
     8821 aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga
     8881 ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa
     8941 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     9001 ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
     9061 tgattggcag aactacacac cagggccagg gatcagatat ccactgacct ttggatggtg
     9121 ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag gagagaacac
     9181 ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtattaga
     9241 gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc atccggactg
     9301 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     9361 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     9421 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     9481 tagcagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     9541 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     9601 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     9661 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     9721 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     9781 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     9841 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     9901 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     9961 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
    10021 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
    10081 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
    10141 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
    10201 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
    10261 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
    10321 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
    10381 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
    10441 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
    10501 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
    10561 gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca
    10621 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag
    10681 ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg
    10741 accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg
    10801 gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg
    10861 gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc
    10921 acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg
    10981 cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag
    11041 gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
    11101 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
    11161 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
    11221 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
    11281 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
    11341 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
    11401 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
    11461 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
    11521 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
    11581 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
    11641 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
    11701 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
    11761 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
    11821 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
    11881 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
    11941 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
    12001 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
    12061 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
    12121 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
    12181 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
    12241 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
    12301 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
    12361 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
    12421 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
    12481 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
    12541 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
    12601 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
    12661 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
    12721 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
    12781 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
    12841 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
    12901 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
    12961 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
    13021 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
    13081 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
    13141 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
    13201 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
    13261 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
    13321 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
    13381 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
    13441 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
//
