LOCUS       Exported               14932 bp DNA     circular SYN 13-DEC-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 14932)
  AUTHORS   Michael Wehr
  TITLE     Direct Submission
  JOURNAL   Exported Jun 22, 2021 from SnapGene Viewer 5.3.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14932
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     CDS             1..66
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             73..93
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             103..4206
                     /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"
     primer_bind     462..483
                     /label=O1457
                     /note="dCas9_7_fwd"
     primer_bind     791..811
                     /label=O1440
                     /note="dCas9_2_fwd"
     primer_bind     complement(841..863)
                     /label=O1439
                     /note="dCas9_1_rev"
     primer_bind     1566..1589
                     /label=O1441
                     /note="dCas9_3_fwd"
     primer_bind     2154..2175
                     /label=O1458
                     /note="dCas9_8_fwd"
     primer_bind     2369..2393
                     /label=O1442
                     /note="dCas9_4_fwd"
     primer_bind     3161..3184
                     /label=O1443
                     /note="dCas9_5_fwd"
     primer_bind     3944..3963
                     /label=O1444
                     /note="dCas9_6_fwd"
     primer_bind     complement(4211..4221)
                     /label=O1439
                     /note="dCas9_1_rev"
     CDS             4219..4239
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     4240..4303
                     /label=O3579
     CDS             4246..4266
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4282..4581
                     /codon_start=1
                     /label=ZIM3-KRAB
                     /translation="MNNSQGRVTFEDVTVNFTQGEWQRLNPEQRNLYRDVMLENYSNLV
                     SVGQGETTKPDVILRLEQGKEPWLEEEEVLGSGRAEKNGDIGGQIWKPKDVKESL"
     misc_structure  4303..4521
                     /label=KRAB domain
     primer_bind     complement(4557..4590)
                     /label=O3561
     CDS             4588..4641
                     /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             4642..5241
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4642..4661)
                     /label=O3580 Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4778..4797
                     /label=O1718
                     /note="PuroR_R"
     primer_bind     4956..4975
                     /label=O1717
                     /note="PuroR_F"
     primer_bind     5138..5158
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    5295..5328
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    5353..5941
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5382..5407)
                     /label=O349
                     /note="pAi9_as"
     primer_bind     complement(5406..5426)
                     /label=O1105
                     /note="WPRE-R"
     primer_bind     complement(5406..5426)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(5420..5439)
                     /label=O1066
                     /note="WPRE_as"
     CDS             complement(5824..5835)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5944..5960)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5945..5961)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6466..6646
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6672..6689)
                     /label=O1104
                     /note="BGH Reverse"
     primer_bind     complement(6672..6689)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     primer_bind     complement(6675..6694)
                     /label=O32
                     /note="BGHrev"
     polyA_signal    6678..6902
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6948..7376
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7035..7054)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7245..7266
                     /label=O1095
                     /note="F1ori-F"
     primer_bind     7245..7266
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7385..7405)
                     /label=O236
                     /note="pBABE 3?"
     primer_bind     complement(7385..7405)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7390..7719
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(7403..7421)
                     /label=O352
                     /note="pLenti6_rev"
     primer_bind     complement(7472..7493)
                     /label=O352
                     /note="pLenti6_rev"
     rep_origin      7570..7705
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7632..7651
                     /label=O1101
                     /note="SV40pro-F"
     primer_bind     7632..7651
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7668..7688
                     /label=O37
                     /note="SV40 promotor forw"
     promoter        7767..7814
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7833..8207
                     /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"
     primer_bind     complement(8335..8354)
                     /label=O342
                     /note="pRK5_as"
     polyA_signal    8337..8458
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8374..8393)
                     /label=O1100
                     /note="SV40pA-R"
     primer_bind     complement(8374..8393)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(8380..8397)
                     /label=O59
                     /note="SV40_rev"
     primer_bind     8380..8397
                     /label=O54
                     /note="SV40_se"
     primer_bind     8428..8447
                     /label=O1109
                     /note="EBV Reverse"
     primer_bind     8428..8447
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8507..8523)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8507..8523)
                     /label=O56
                     /note="M13 reverse"
     primer_bind     complement(8507..8523)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8519..8542)
                     /label=O1126
                     /note="M13 rev (-48)"
     primer_bind     complement(8520..8542)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8531..8547
                     /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(8555..8585)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8600..8621
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8738..8755)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8909..9497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8989..9008)
                     /label=O1094
                     /note="pBR322ori-F"
     primer_bind     complement(8989..9008)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9668..10528)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(9711..9728)
                     /label=O139
                     /note="AmpR_3' fwd"
     primer_bind     10291..10310
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     10361..10378
                     /label=O140
                     /note="AmpR_5' rev"
     primer_bind     complement(10461..10478)
                     /label=O138
                     /note="AmpR_5' fwd"
     promoter        complement(10529..10633)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10708..10727)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        10899..11278
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     10924..10941
                     /label=O136
                     /note="CMVprom_5' fwd"
     primer_bind     complement(11059..11076)
                     /label=O137
                     /note="CMVprom_5' rev"
     primer_bind     11098..11123
                     /label=O353
                     /note="pLenti6.2/V5_fwd"
     promoter        11280..11478
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11433..11453
                     /label=O1103
                     /note="CMV Forward"
     primer_bind     11433..11453
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     11435..11452
                     /label=O53
                     /note="pCMV_se"
     LTR             11496..11676
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11723..11848
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12341..12574
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    13101..13218
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    13262..13295
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_RNA        complement(13306..13377)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     13311..13332
                     /label=O3094
                     /note="sgRNA_colonyPCR_rev"
     misc_feature    complement(13382..13401)
                     /label=sgTAOK2_CRISPRi_3
     promoter        complement(13411..13651)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     complement(13455..13481)
                     /label=O3095
                     /note="sgRNA_colonyPCR_fwd"
     primer_bind     complement(13461..13480)
                     /label=O227
                     /note="pLKO.1 puro fwd"
     primer_bind     complement(13461..13480)
                     /label=LKO.1 5' (O227)
                     /note="Human U6 promoter, forward primer"
     primer_bind     13584..13607
                     /label=O2146
                     /note="pLKO.1_shRNA_rev"
     primer_bind     complement(13584..13607)
                     /label=O223
                     /note="pLKO.1_shRNA_fwd"
     primer_bind     complement(13629..13650)
                     /label=O2050
                     /note="hU6_fwd"
     primer_bind     complement(13631..13651)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     13666..13685
                     /label=O3098
                     /note="hUbC_SalI_fwd"
     primer_bind     13668..13686
                     /label=O3445
     promoter        13684..14895
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     14732..14751
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     primer_bind     complement(14889..14910)
                     /label=O3446
     primer_bind     complement(14896..14914)
                     /label=O3099
                     /note="hUbC_MluI_rev"
     regulatory      join(14924..14932,1)
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat
       61 gacaagatgg cccccaagaa gaagaggaag gtgggccgcg gaatggacaa gaagtactcc
      121 attgggctcg ccatcggcac aaacagcgtc ggctgggccg tcattacgga cgagtacaag
      181 gtgccgagca aaaaattcaa agttctgggc aataccgatc gccacagcat aaagaagaac
      241 ctcattggcg ccctcctgtt cgactccggg gaaaccgccg aagccacgcg gctcaaaaga
      301 acagcacggc gcagatatac ccgcagaaag aatcggatct gctacctgca ggagatcttt
      361 agtaatgaga tggctaaggt ggatgactct ttcttccata ggctggagga gtcctttttg
      421 gtggaggagg ataaaaagca cgagcgccac ccaatctttg gcaatatcgt ggacgaggtg
      481 gcgtaccatg aaaagtaccc aaccatatat catctgagga agaagcttgt agacagtact
      541 gataaggctg acttgcggtt gatctatctc gcgctggcgc atatgatcaa atttcgggga
      601 cacttcctca tcgaggggga cctgaaccca gacaacagcg atgtcgacaa actctttatc
      661 caactggttc agacttacaa tcagcttttc gaagagaacc cgatcaacgc atccggagtt
      721 gacgccaaag caatcctgag cgctaggctg tccaaatccc ggcggctcga aaacctcatc
      781 gcacagctcc ctggggagaa gaagaacggc ctgtttggta atcttatcgc cctgtcactc
      841 gggctgaccc ccaactttaa atctaacttc gacctggccg aagatgccaa gcttcaactg
      901 agcaaagaca cctacgatga tgatctcgac aatctgctgg cccagatcgg cgaccagtac
      961 gcagaccttt ttttggcggc aaagaacctg tcagacgcca ttctgctgag tgatattctg
     1021 cgagtgaaca cggagatcac caaagctccg ctgagcgcta gtatgatcaa gcgctatgat
     1081 gagcaccacc aagacttgac tttgctgaag gcccttgtca gacagcaact gcctgagaag
     1141 tacaaggaaa ttttcttcga tcagtctaaa aatggctacg ccggatacat tgacggcgga
     1201 gcaagccagg aggaatttta caaatttatt aagcccatct tggaaaaaat ggacggcacc
     1261 gaggagctgc tggtaaagct taacagagaa gatctgttgc gcaaacagcg cactttcgac
     1321 aatggaagca tcccccacca gattcacctg ggcgaactgc acgctatcct caggcggcaa
     1381 gaggatttct accccttttt gaaagataac agggaaaaga ttgagaaaat cctcacattt
     1441 cggataccct actatgtagg ccccctcgcc cggggaaatt ccagattcgc gtggatgact
     1501 cgcaaatcag aagagaccat cactccctgg aacttcgagg aagtcgtgga taagggggcc
     1561 tctgcccagt ccttcatcga aaggatgact aactttgata aaaatctgcc taacgaaaag
     1621 gtgcttccta aacactctct gctgtacgag tacttcacag tttataacga gctcaccaag
     1681 gtcaaatacg tcacagaagg gatgagaaag ccagcattcc tgtctggaga gcagaagaaa
     1741 gctatcgtgg acctcctctt caagacgaac cggaaagtta ccgtgaaaca gctcaaagaa
     1801 gactatttca aaaagattga atgtttcgac tctgttgaaa tcagcggagt ggaggatcgc
     1861 ttcaacgcat ccctgggaac gtatcacgat ctcctgaaaa tcattaaaga caaggacttc
     1921 ctggacaatg aggagaacga ggacattctt gaggacattg tcctcaccct tacgttgttt
     1981 gaagataggg agatgattga agaacgcttg aaaacttacg ctcatctctt cgacgacaaa
     2041 gtcatgaaac agctcaagag gcgccgatat acaggatggg ggcggctgtc aagaaaactg
     2101 atcaatggga tccgagacaa gcagagtgga aagacaatcc tggattttct taagtccgat
     2161 ggatttgcca accggaactt catgcagttg atccatgatg actctctcac ctttaaggag
     2221 gacatccaga aagcacaagt ttctggccag ggggacagtc ttcacgagca catcgctaat
     2281 cttgcaggta gcccagctat caaaaaggga atactgcaga ccgttaaggt cgtggatgaa
     2341 ctcgtcaaag taatgggaag gcataagccc gagaatatcg ttatcgagat ggcccgagag
     2401 aaccaaacta cccagaaggg acagaagaac agtagggaaa ggatgaagag gattgaagag
     2461 ggtataaaag aactggggtc ccaaatcctt aaggaacacc cagttgaaaa cacccagctt
     2521 cagaatgaga agctctacct gtactacctg cagaacggca gggacatgta cgtggatcag
     2581 gaactggaca tcaatcggct ctccgactac gacgtggatg ccatcgtgcc ccagtctttt
     2641 ctcaaagatg attctattga taataaagtg ttgacaagat ccgataaaaa tagagggaag
     2701 agtgataacg tcccctcaga agaagttgtc aagaaaatga aaaattattg gcggcagctg
     2761 ctgaacgcca aactgatcac acaacggaag ttcgataatc tgactaaggc tgaacgaggt
     2821 ggcctgtctg agttggataa agccggcttc atcaaaaggc agcttgttga gacacgccag
     2881 atcaccaagc acgtggccca aattctcgat tcacgcatga acaccaagta cgatgaaaat
     2941 gacaaactga ttcgagaggt gaaagttatt actctgaagt ctaagctggt ctcagatttc
     3001 agaaaggact ttcagtttta taaggtgaga gagatcaaca attaccacca tgcgcatgat
     3061 gcctacctga atgcagtggt aggcactgca cttatcaaaa aatatcccaa gcttgaatct
     3121 gaatttgttt acggagacta taaagtgtac gatgttagga aaatgatcgc aaagtctgag
     3181 caggaaatag gcaaggccac cgctaagtac ttcttttaca gcaatattat gaattttttc
     3241 aagaccgaga ttacactggc caatggagag attcggaagc gaccacttat cgaaacaaac
     3301 ggagaaacag gagaaatcgt gtgggacaag ggtagggatt tcgcgacagt ccggaaggtc
     3361 ctgtccatgc cgcaggtgaa catcgttaaa aagaccgaag tacagaccgg aggcttctcc
     3421 aaggaaagta tcctcccgaa aaggaacagc gacaagctga tcgcacgcaa aaaagattgg
     3481 gaccccaaga aatacggcgg attcgattct cctacagtcg cttacagtgt actggttgtg
     3541 gccaaagtgg agaaagggaa gtctaaaaaa ctcaaaagcg tcaaggaact gctgggcatc
     3601 acaatcatgg agcgatcaag cttcgaaaaa aaccccatcg actttctcga ggcgaaagga
     3661 tataaagagg tcaaaaaaga cctcatcatt aagcttccca agtactctct ctttgagctt
     3721 gaaaacggcc ggaaacgaat gctcgctagt gcgggcgagc tgcagaaagg taacgagctg
     3781 gcactgccct ctaaatacgt taatttcttg tatctggcca gccactatga aaagctcaaa
     3841 gggtctcccg aagataatga gcagaagcag ctgttcgtgg aacaacacaa acactacctt
     3901 gatgagatca tcgagcaaat aagcgaattc tccaaaagag tgatcctcgc cgacgctaac
     3961 ctcgataagg tgctttctgc ttacaataag cacagggata agcccatcag ggagcaggca
     4021 gaaaacatta tccacttgtt tactctgacc aacttgggcg cgcctgcagc cttcaagtac
     4081 ttcgacacca ccatagacag aaagcggtac acctctacaa aggaggtcct ggacgccaca
     4141 ctgattcatc agtcaattac ggggctctat gaaacaagaa tcgacctctc tcagctcggt
     4201 ggagacagca gggctgaccc caagaagaag aggaaggtgg ctagccctaa gaaaaagcgg
     4261 aaagtggtta acggaggcgg aatgaacaat tcccagggaa gagtgacctt cgaggatgtc
     4321 actgtgaact tcacccaggg ggagtggcag cggctgaatc ccgaacagag aaacttgtac
     4381 agggatgtga tgctggagaa ttacagcaac cttgtctctg tgggacaagg ggaaaccacc
     4441 aaacccgatg tgatcttgag gttggaacaa ggaaaggagc catggttgga ggaagaggaa
     4501 gtgctgggaa gtggccgtgc agaaaaaaat ggggacattg gagggcagat ttggaagcca
     4561 aaggatgtga aagagagtct cgctagcgag ggcagaggaa gtcttctaac atgcggtgac
     4621 gtggaggaga atcccggccc tatgaccgag tacaagccca cggtgcgcct cgccacccgc
     4681 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     4741 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     4801 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     4861 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     4921 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     4981 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt gtcgcccgac
     5041 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     5101 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     5161 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     5221 acccgcaagc ccggtgcctg accagcacac tggcggccgt tactagcttc tgcagcacga
     5281 ccggttgata atagataact tcgtatagca tacattatac gaagttatga attcgatatc
     5341 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     5401 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     5461 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     5521 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     5581 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     5641 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     5701 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     5761 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     5821 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     5881 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     5941 catcgatacc gtcgacctcg agacctagaa aaacatggag caatcacaag tagcaataca
     6001 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt
     6061 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     6121 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     6181 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattggca gaactacaca
     6241 ccagggccag ggatcagata tccactgacc tttggatggt gctacaagct agtaccagtt
     6301 gagcaagaga aggtagaaga agccaatgaa ggagagaaca cccgcttgtt acaccctgtg
     6361 agcctgcatg ggatggatga cccggagaga gaagtattag agtggaggtt tgacagccgc
     6421 ctagcatttc atcacatggc ccgagagctg catccggact gtactgggtc tctctggtta
     6481 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     6541 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     6601 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagggc ccgtttaaac
     6661 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     6721 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     6781 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     6841 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     6901 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     6961 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     7021 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     7081 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     7141 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     7201 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     7261 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     7321 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     7381 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     7441 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     7501 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     7561 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     7621 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     7681 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     7741 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     7801 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     7861 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     7921 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     7981 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     8041 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     8101 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     8161 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     8221 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     8281 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     8341 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     8401 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     8461 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     8521 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     8581 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     8641 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     8701 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     8761 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     8821 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     8881 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     8941 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     9001 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     9061 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     9121 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     9181 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     9241 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     9301 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     9361 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     9421 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     9481 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     9541 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     9601 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     9661 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     9721 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     9781 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     9841 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     9901 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     9961 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
    10021 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
    10081 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
    10141 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
    10201 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
    10261 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
    10321 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
    10381 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
    10441 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
    10501 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
    10561 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
    10621 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
    10681 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
    10741 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
    10801 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
    10861 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
    10921 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
    10981 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
    11041 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
    11101 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
    11161 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
    11221 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
    11281 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
    11341 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
    11401 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
    11461 ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct
    11521 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
    11581 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
    11641 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
    11701 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
    11761 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
    11821 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
    11881 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
    11941 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
    12001 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
    12061 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
    12121 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
    12181 gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
    12241 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
    12301 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
    12361 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
    12421 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
    12481 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
    12541 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
    12601 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
    12661 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
    12721 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
    12781 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
    12841 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
    12901 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
    12961 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
    13021 gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa
    13081 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
    13141 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
    13201 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttggttaatt
    13261 aataacttcg tatagcatac attatacgaa gttattaaaa aaaaagcacc gactcggtgc
    13321 cactttttca agttgataac ggactagcct tattttaact tgctatttct agctctaaaa
    13381 cggtctcgtc ccggtctccc tcggtgtttc gtcctttcca caagatatat aaagccaaga
    13441 aatcgaaata ctttcaagtt acggtaagca tatgatagtc cattttaaaa cataatttta
    13501 aaactgcaaa ctacccaaga aattattact ttctacgtca cgtattttgt actaatatct
    13561 ttgtgtttac agtcaaatta attccaatta tctctctaac agccttgtat cgtatatgca
    13621 aatatgaagg aatcatggga aataggccct cttaattaat taacccgtgt cggctccaga
    13681 tctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcctc acggcgagcg
    13741 ctgccacgtc agacgaaggg cgcagcgagc gtcctgatcc ttccgcccgg acgctcagga
    13801 cagcggcccg ctgctcataa gactcggcct tagaacccca gtatcagcag aaggacattt
    13861 taggacggga cttgggtgac tctagggcac tggttttctt tccagagagc ggaacaggcg
    13921 aggaaaagta gtcccttctc ggcgattctg cggagggatc tccgtggggc ggtgaacgcc
    13981 gatgattata taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg
    14041 ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg
    14101 gctggccggg gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt gtggagagac
    14161 cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg gttgggggga
    14221 gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg
    14281 tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag gtcttgaggc
    14341 cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctggggcacc atctggggac
    14401 cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg cgggggcggc
    14461 agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt
    14521 gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacct gccggtaggt
    14581 gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta ggctctcctg
    14641 aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca gtttctttgg
    14701 tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg
    14761 gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta atcatttggg
    14821 tcaatatgta attttcagtg ttagactagt aaattgtccg ctaaattctg gccgtttttg
    14881 gcttttttgt tagacgaagc ttgggctgca ggtcgactct agagccacca tg
//
