LOCUS       Exported               15111 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Co-expresses human optimized S. pyogenes dCas9 fused to two copies 
            of VP64 and GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pLV hUbC-VP64 dCas9 VP64-T2A-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15111)
  AUTHORS   Kabadi AM, Ousterout DG, Hilton IB, Gersbach CA
  TITLE     Multiplex CRISPR/Cas9-based genome engineering from a single 
            lentiviral vector.
  JOURNAL   Nucleic Acids Res. 2014 Aug 13. pii: gku749.
  PUBMED    25122746
REFERENCE   2  (bases 1 to 15111)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..15111
                     /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"
     protein_bind    2365..2398
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(2385..2711)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGATTVSY
                     HNFV"
     primer_bind     2561..2583
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2575..2592
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2576..2592
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2602..2621
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2602..2620
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2668..2688)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2668..2686)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2707..2723)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2731..2747
                     /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(2755..2785)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2800..2821
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2898..4109
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3946..3965
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      4138..4147
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4147..4212
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4228..4374
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDM"
     CDS             4381..4401
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4411..8514
                     /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 H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8527..8547
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8560..8709
                     /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             8719..8772
                     /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             8779..9492
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             8779..9492
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(8821..8842)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(9082..9101)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9429..9450
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    9508..9541
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    9566..10154
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9619..9639)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10037..10048)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10157..10173)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10158..10174)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10679..10859
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10885..10902)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10891..11115
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11161..11589
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11248..11267)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11458..11479
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11598..11618)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11603..11932
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11783..11918
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11845..11864
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11980..12027
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12046..12420
                     /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    12550..12671
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12587..12606)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12641..12660
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12720..12736)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    12744..12760
                     /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(12768..12798)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12813..12834
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12951..12968)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13122..13710)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13202..13221)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(13881..14741)
                     /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     14504..14523
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14742..14846)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14921..14940)
                     /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 ttaaataact tcgtatagca tacattatac gaagttatga
     2401 taagagacgg tggttgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
     2461 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     2521 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     2581 acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg taccgggccc
     2641 cccctcgagg tcctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     2701 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     2761 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2821 aattgcgttg cgctcactgc ccgctttcca ctgcatgacg tctccacaat taattaaccc
     2881 gtgtcggctc cagatctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct
     2941 cctcacggcg agcgctgcca cgtcagacga agggcgcagc gagcgtcctg atccttccgc
     3001 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
     3061 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
     3121 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
     3181 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
     3241 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
     3301 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
     3361 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
     3421 gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct
     3481 tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc
     3541 caaggtcttg aggccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggg
     3601 caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct
     3661 gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg
     3721 cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc
     3781 acctgccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag
     3841 ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc
     3901 gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt
     3961 gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa
     4021 tgtaatcatt tgggtcaata tgtaattttc agtgttagac tagtaaattg tccgctaaat
     4081 tctggccgtt tttggctttt ttgttagacg aagcttgggc tgcaggtcga ctctagagcc
     4141 accatggact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
     4201 gacgatgaca agcacggtcc gcgggctgac gcattggacg attttgatct ggatatgctg
     4261 ggaagtgacg ccctcgatga ttttgacctt gacatgcttg gttcggatgc ccttgatgac
     4321 tttgacctcg acatgctcgg cagtgacgcc cttgatgatt tcgacctgga catggttaac
     4381 cccaagaaga agaggaaggt gggccgcgga atggacaaga agtactccat tgggctcgcc
     4441 atcggcacaa acagcgtcgg ctgggccgtc attacggacg agtacaaggt gccgagcaaa
     4501 aaattcaaag ttctgggcaa taccgatcgc cacagcataa agaagaacct cattggcgcc
     4561 ctcctgttcg actccgggga aaccgccgaa gccacgcggc tcaaaagaac agcacggcgc
     4621 agatataccc gcagaaagaa tcggatctgc tacctgcagg agatctttag taatgagatg
     4681 gctaaggtgg atgactcttt cttccatagg ctggaggagt cctttttggt ggaggaggat
     4741 aaaaagcacg agcgccaccc aatctttggc aatatcgtgg acgaggtggc gtaccatgaa
     4801 aagtacccaa ccatatatca tctgaggaag aagcttgtag acagtactga taaggctgac
     4861 ttgcggttga tctatctcgc gctggcgcat atgatcaaat ttcggggaca cttcctcatc
     4921 gagggggacc tgaacccaga caacagcgat gtcgacaaac tctttatcca actggttcag
     4981 acttacaatc agcttttcga agagaacccg atcaacgcat ccggagttga cgccaaagca
     5041 atcctgagcg ctaggctgtc caaatcccgg cggctcgaaa acctcatcgc acagctccct
     5101 ggggagaaga agaacggcct gtttggtaat cttatcgccc tgtcactcgg gctgaccccc
     5161 aactttaaat ctaacttcga cctggccgaa gatgccaagc ttcaactgag caaagacacc
     5221 tacgatgatg atctcgacaa tctgctggcc cagatcggcg accagtacgc agaccttttt
     5281 ttggcggcaa agaacctgtc agacgccatt ctgctgagtg atattctgcg agtgaacacg
     5341 gagatcacca aagctccgct gagcgctagt atgatcaagc gctatgatga gcaccaccaa
     5401 gacttgactt tgctgaaggc ccttgtcaga cagcaactgc ctgagaagta caaggaaatt
     5461 ttcttcgatc agtctaaaaa tggctacgcc ggatacattg acggcggagc aagccaggag
     5521 gaattttaca aatttattaa gcccatcttg gaaaaaatgg acggcaccga ggagctgctg
     5581 gtaaagctta acagagaaga tctgttgcgc aaacagcgca ctttcgacaa tggaagcatc
     5641 ccccaccaga ttcacctggg cgaactgcac gctatcctca ggcggcaaga ggatttctac
     5701 ccctttttga aagataacag ggaaaagatt gagaaaatcc tcacatttcg gataccctac
     5761 tatgtaggcc ccctcgcccg gggaaattcc agattcgcgt ggatgactcg caaatcagaa
     5821 gagaccatca ctccctggaa cttcgaggaa gtcgtggata agggggcctc tgcccagtcc
     5881 ttcatcgaaa ggatgactaa ctttgataaa aatctgccta acgaaaaggt gcttcctaaa
     5941 cactctctgc tgtacgagta cttcacagtt tataacgagc tcaccaaggt caaatacgtc
     6001 acagaaggga tgagaaagcc agcattcctg tctggagagc agaagaaagc tatcgtggac
     6061 ctcctcttca agacgaaccg gaaagttacc gtgaaacagc tcaaagaaga ctatttcaaa
     6121 aagattgaat gtttcgactc tgttgaaatc agcggagtgg aggatcgctt caacgcatcc
     6181 ctgggaacgt atcacgatct cctgaaaatc attaaagaca aggacttcct ggacaatgag
     6241 gagaacgagg acattcttga ggacattgtc ctcaccctta cgttgtttga agatagggag
     6301 atgattgaag aacgcttgaa aacttacgct catctcttcg acgacaaagt catgaaacag
     6361 ctcaagaggc gccgatatac aggatggggg cggctgtcaa gaaaactgat caatgggatc
     6421 cgagacaagc agagtggaaa gacaatcctg gattttctta agtccgatgg atttgccaac
     6481 cggaacttca tgcagttgat ccatgatgac tctctcacct ttaaggagga catccagaaa
     6541 gcacaagttt ctggccaggg ggacagtctt cacgagcaca tcgctaatct tgcaggtagc
     6601 ccagctatca aaaagggaat actgcagacc gttaaggtcg tggatgaact cgtcaaagta
     6661 atgggaaggc ataagcccga gaatatcgtt atcgagatgg cccgagagaa ccaaactacc
     6721 cagaagggac agaagaacag tagggaaagg atgaagagga ttgaagaggg tataaaagaa
     6781 ctggggtccc aaatccttaa ggaacaccca gttgaaaaca cccagcttca gaatgagaag
     6841 ctctacctgt actacctgca gaacggcagg gacatgtacg tggatcagga actggacatc
     6901 aatcggctct ccgactacga cgtggatgcc atcgtgcccc agtcttttct caaagatgat
     6961 tctattgata ataaagtgtt gacaagatcc gataaaaata gagggaagag tgataacgtc
     7021 ccctcagaag aagttgtcaa gaaaatgaaa aattattggc ggcagctgct gaacgccaaa
     7081 ctgatcacac aacggaagtt cgataatctg actaaggctg aacgaggtgg cctgtctgag
     7141 ttggataaag ccggcttcat caaaaggcag cttgttgaga cacgccagat caccaagcac
     7201 gtggcccaaa ttctcgattc acgcatgaac accaagtacg atgaaaatga caaactgatt
     7261 cgagaggtga aagttattac tctgaagtct aagctggtct cagatttcag aaaggacttt
     7321 cagttttata aggtgagaga gatcaacaat taccaccatg cgcatgatgc ctacctgaat
     7381 gcagtggtag gcactgcact tatcaaaaaa tatcccaagc ttgaatctga atttgtttac
     7441 ggagactata aagtgtacga tgttaggaaa atgatcgcaa agtctgagca ggaaataggc
     7501 aaggccaccg ctaagtactt cttttacagc aatattatga attttttcaa gaccgagatt
     7561 acactggcca atggagagat tcggaagcga ccacttatcg aaacaaacgg agaaacagga
     7621 gaaatcgtgt gggacaaggg tagggatttc gcgacagtcc ggaaggtcct gtccatgccg
     7681 caggtgaaca tcgttaaaaa gaccgaagta cagaccggag gcttctccaa ggaaagtatc
     7741 ctcccgaaaa ggaacagcga caagctgatc gcacgcaaaa aagattggga ccccaagaaa
     7801 tacggcggat tcgattctcc tacagtcgct tacagtgtac tggttgtggc caaagtggag
     7861 aaagggaagt ctaaaaaact caaaagcgtc aaggaactgc tgggcatcac aatcatggag
     7921 cgatcaagct tcgaaaaaaa ccccatcgac tttctcgagg cgaaaggata taaagaggtc
     7981 aaaaaagacc tcatcattaa gcttcccaag tactctctct ttgagcttga aaacggccgg
     8041 aaacgaatgc tcgctagtgc gggcgagctg cagaaaggta acgagctggc actgccctct
     8101 aaatacgtta atttcttgta tctggccagc cactatgaaa agctcaaagg gtctcccgaa
     8161 gataatgagc agaagcagct gttcgtggaa caacacaaac actaccttga tgagatcatc
     8221 gagcaaataa gcgaattctc caaaagagtg atcctcgccg acgctaacct cgataaggtg
     8281 ctttctgctt acaataagca cagggataag cccatcaggg agcaggcaga aaacattatc
     8341 cacttgttta ctctgaccaa cttgggcgcg cctgcagcct tcaagtactt cgacaccacc
     8401 atagacagaa agcggtacac ctctacaaag gaggtcctgg acgccacact gattcatcag
     8461 tcaattacgg ggctctatga aacaagaatc gacctctctc agctcggtgg agacagcagg
     8521 gctgacccca agaagaagag gaaggtggct agccgcgccg acgcgctgga cgatttcgat
     8581 ctcgacatgc tgggttctga tgccctcgat gactttgacc tggatatgtt gggaagcgac
     8641 gcattggatg actttgatct ggacatgctc ggctccgatg ctctggacga tttcgatctc
     8701 gatatgttaa tcgctagcga gggcagagga agtcttctaa catgcggtga cgtggaggag
     8761 aatcccggcc ctggtaccgt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     8821 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     8881 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     8941 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     9001 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     9061 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     9121 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     9181 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     9241 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     9301 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     9361 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     9421 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     9481 gagctgtaca agaccggttg ataatagata acttcgtata gcatacatta tacgaagtta
     9541 tgaattcgat atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt
     9601 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     9661 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     9721 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     9781 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     9841 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     9901 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     9961 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
    10021 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
    10081 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
    10141 ggccgcctcc ccgcatcgat accgtcgacc tcgagaccta gaaaaacatg gagcaatcac
    10201 aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga
    10261 ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc
    10321 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
    10381 acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact tccctgattg
    10441 gcagaactac acaccagggc cagggatcag atatccactg acctttggat ggtgctacaa
    10501 gctagtacca gttgagcaag agaaggtaga agaagccaat gaaggagaga acacccgctt
    10561 gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat tagagtggag
    10621 gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg actgtactgg
    10681 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
    10741 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
    10801 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
    10861 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
    10921 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
    10981 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
    11041 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
    11101 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
    11161 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
    11221 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
    11281 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
    11341 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
    11401 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
    11461 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
    11521 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
    11581 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
    11641 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
    11701 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
    11761 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
    11821 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
    11881 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
    11941 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
    12001 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc
    12061 agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac
    12121 cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac
    12181 gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg
    12241 gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac
    12301 ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag
    12361 ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga
    12421 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
    12481 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
    12541 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
    12601 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
    12661 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
    12721 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
    12781 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
    12841 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
    12901 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
    12961 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
    13021 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
    13081 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
    13141 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
    13201 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
    13261 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
    13321 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
    13381 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
    13441 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
    13501 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
    13561 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
    13621 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
    13681 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
    13741 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
    13801 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
    13861 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
    13921 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
    13981 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
    14041 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
    14101 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
    14161 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
    14221 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
    14281 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
    14341 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
    14401 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
    14461 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
    14521 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
    14581 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
    14641 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
    14701 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
    14761 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
    14821 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
    14881 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
    14941 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
    15001 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
    15061 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt t
//
