LOCUS       Exported               13905 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  lentiviral expression vector for dCas9 with Blasticidin selectable 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    pGH125_dCas9-Blast
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13905)
  AUTHORS   Hess GT, Fresard L, Han K, Lee CH, Li A, Cimprich KA, Montgomery SB,
            Bassik MC
  TITLE     Directed evolution using dCas9-targeted somatic hypermutation in 
            mammalian cells.
  JOURNAL   Nat Methods. 2016 Oct 31. doi: 10.1038/nmeth.4038.
  PUBMED    27798611
REFERENCE   2  (bases 1 to 13905)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..13905
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /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    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2450..3628
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2681..3619
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3576..3596
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3644..3691
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             3692..7792
                     /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             7847..7867
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7883..7936
                     /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             7937..8335
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8360..8948
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8413..8433)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8831..8842)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8951..8967)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8952..8968)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9473..9653
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9679..9696)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9685..9909
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9955..10383
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10042..10061)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10252..10273
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10392..10412)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10397..10726
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10577..10712
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10639..10658
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10774..10821
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10840..11214
                     /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    11344..11465
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11381..11400)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11435..11454
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11514..11530)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11514..11530)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11527..11549)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11538..11554
                     /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(11562..11592)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11607..11628
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11745..11762)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11916..12504)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11996..12015)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(12675..13535)
                     /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     13298..13317
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13536..13640)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(13715..13734)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttagctagct gcaaagatgg ataaagtttt aaacagagag
     2401 gaatctttgc agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc
     2461 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     2521 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     2581 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     2641 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc
     2701 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg
     2761 gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag
     2821 gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg
     2881 gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc
     2941 tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct
     3001 tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga
     3061 cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc
     3121 gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc
     3181 gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc
     3241 ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc
     3301 gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt
     3361 cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag
     3421 cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca
     3481 cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga
     3541 atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag
     3601 tttttttctt ccatttcagg tgtcgtgacg tacggccacc atgaaaaggc cggcggccac
     3661 gaaaaaggcc ggccaggcaa aaaagaaaaa ggacaagaag tacagcatcg gcctggccat
     3721 cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa
     3781 attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct
     3841 gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag
     3901 atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc
     3961 caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa
     4021 gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa
     4081 gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct
     4141 gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga
     4201 gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac
     4261 ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat
     4321 cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg
     4381 cgagaagaag aatggcctgt tcggcaacct gattgccctg agcctgggcc tgacccccaa
     4441 cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta
     4501 cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct
     4561 ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga
     4621 gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga
     4681 cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt
     4741 cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga
     4801 gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt
     4861 gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc
     4921 ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc
     4981 attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta
     5041 cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga
     5101 aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt
     5161 catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca
     5221 cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac
     5281 cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct
     5341 gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa
     5401 aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct
     5461 gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga
     5521 aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat
     5581 gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct
     5641 gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg
     5701 ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag
     5761 aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc
     5821 ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc
     5881 cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat
     5941 gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca
     6001 gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct
     6061 gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct
     6121 gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa
     6181 ccggctgtcc gactacgatg tggaccacat cgtgcctcag agctttctga aggacgactc
     6241 catcgacaac aaggtgctga ccagaagcga caaggcccgg ggcaagagcg acaacgtgcc
     6301 ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct
     6361 gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact
     6421 ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt
     6481 ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg
     6541 ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca
     6601 gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc
     6661 cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg
     6721 cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa
     6781 ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac
     6841 cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga
     6901 gatcgtgtgg gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca
     6961 agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct
     7021 gcccaagagg aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta
     7081 cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa
     7141 gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag
     7201 aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa
     7261 aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa
     7321 gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa
     7381 atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga
     7441 taatgagcag aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga
     7501 gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct
     7561 gtccgcctac aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca
     7621 cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat
     7681 cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag
     7741 catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acagcgctgg
     7801 aggaggtgga agcggaggag gaggaagcgg aggaggaggt agcggaccta agaaaaagag
     7861 gaaggtggcg gccgctggat ccgagggcag aggaagtctg ctaacatgcg gtgacgtcga
     7921 ggagaatcct ggcccaatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag
     7981 agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc
     8041 tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc
     8101 ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg
     8161 tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca
     8221 ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc
     8281 gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaagaatt
     8341 cgatatcaag cttatcggta atcaacctct ggattacaaa atttgtgaaa gattgactgg
     8401 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     8461 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     8521 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     8581 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     8641 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     8701 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     8761 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     8821 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     8881 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     8941 ctccccgcat cgataccgtc gacctcgaga cctagaaaaa catggagcaa tcacaagtag
     9001 caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt
     9061 gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga
     9121 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
     9181 acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg attggcagaa
     9241 ctacacacca gggccaggga tcagatatcc actgaccttt ggatggtgct acaagctagt
     9301 accagttgag caagagaagg tagaagaagc caatgaagga gagaacaccc gcttgttaca
     9361 ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtattagagt ggaggtttga
     9421 cagccgccta gcatttcatc acatggcccg agagctgcat ccggactgta ctgggtctct
     9481 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     9541 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     9601 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagggcccg
     9661 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     9721 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     9781 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     9841 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     9901 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     9961 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
    10021 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
    10081 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
    10141 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
    10201 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
    10261 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
    10321 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
    10381 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
    10441 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
    10501 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
    10561 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
    10621 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
    10681 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
    10741 ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat
    10801 atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc
    10861 gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc
    10921 gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc
    10981 ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg
    11041 gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg
    11101 gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc
    11161 ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg
    11221 ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
    11281 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
    11341 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
    11401 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
    11461 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
    11521 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
    11581 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
    11641 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
    11701 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
    11761 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
    11821 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
    11881 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
    11941 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
    12001 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
    12061 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
    12121 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
    12181 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
    12241 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
    12301 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
    12361 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
    12421 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
    12481 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
    12541 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
    12601 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
    12661 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
    12721 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
    12781 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
    12841 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
    12901 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
    12961 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
    13021 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
    13081 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
    13141 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
    13201 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
    13261 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
    13321 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
    13381 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
    13441 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
    13501 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
    13561 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
    13621 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
    13681 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
    13741 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
    13801 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
    13861 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgtt
//
