LOCUS       sequence-388040-       14989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR-inhibition. dCas9-KRAB(ZIM3) with TagBFP for sorting. 
            Improved KRAB domain from ZIM3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14989)
  TITLE     Schmierer unpublished plasmids
REFERENCE   2  (bases 1 to 14989)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: lenti-UCOE-dCas9-BFP-Zim3-KRAB
FEATURES             Location/Qualifiers
     source          1..14989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2380..2497
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3214..4392
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3445..4383
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4340..4360
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4408..4455
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(4456..4479,4480..4482,4483..7038,7039..7041,
                     7042..8556)
                     /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             8611..8631
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8647..9342
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     misc_feature    9681..10269
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9734..9754)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10152..10163)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10272..10288)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10273..10289)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10794..10974
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11000..11017)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11006..11230
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11276..11704
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11363..11382)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11573..11594
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11713..11733)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11718..12047
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11898..12033
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11960..11979
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12095..12142
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12161..12535
                     /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    12665..12786
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12702..12721)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12756..12775
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12835..12851)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12835..12851)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12848..12870)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12859..12875
                     /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(join(12883..12889,12890..12907,12908..12913))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12928..12949
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13066..13083)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13237..13825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13317..13336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13996..14787,14788..14856))
                     /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     14619..14638
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14857..14961)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccagag agggcctcta gagcttggcg
     2341 taactagatc ttgagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2401 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2461 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2521 agatccagtt tgggctagct gcaaagatgg ataaagtttt aaacagagag gaatctttgc
     2581 agctaatgga ccttctaggt cttgaaagga gcctgcaggc gggaggtggt ccctgcagtt
     2641 acgccaatga taacccccgc cagaaaaatc ttagtagcct tccctttttg ttttccgtgc
     2701 cccaactcgg cggattgact cggccccttc cggaaacacc cgaatcaact tctagtcaaa
     2761 ttattgttca cgccgcaatg acccacccct ggcccgcgtc tgtggaactg acccctggtg
     2821 ttcaggagag ttcgctgctg aaagtggtcc caaaggggta ctagttttta agctcccaac
     2881 tccccctccc ccagcgtctg gaggattcca caccctcgca ccgcaggggc gaggaagtgg
     2941 gcggagtccg gttttggcgc cagccgctga ggctgccaag cagaaaagcc accgctgagg
     3001 agactccggt cactgtcctc gccccgcctc ccccttccct ccccttgggg accaccgggc
     3061 gccacgccgc gaacggtaag tgccgcggtc gtcggcgcct ccgccctccc cctagggccc
     3121 caattcccag cgggcgcggc gcgcggcccc gcgcgcagct cccggctccc tcccccttcg
     3181 gatgtggctt gagctgtagg cgcggagcct gcaggctccg gtgcccgtca gtgggcagag
     3241 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc
     3301 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
     3361 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
     3421 aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc
     3481 tttacgggtt atggcccttg cgtgccttga attacttcca cctggctgca gtacgtgatt
     3541 cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga
     3601 gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga
     3661 atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat
     3721 ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa
     3781 gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc
     3841 cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg
     3901 tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg
     3961 ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt
     4021 cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt
     4081 gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc
     4141 acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg
     4201 tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag
     4261 actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag
     4321 tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt
     4381 caggtgtcgt gacgtacggc caccatgaaa aggccggcgg ccacgaaaaa ggccggccag
     4441 gcaaaaaaga aaaaggacaa gaagtacagc atcggcctgg ccatcggcac caactctgtg
     4501 ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc
     4561 aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc
     4621 gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag
     4681 aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc
     4741 ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac
     4801 cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac
     4861 cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg
     4921 gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc
     4981 gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc
     5041 gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg
     5101 agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc
     5161 ctgttcggca acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc
     5221 gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac
     5281 aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg
     5341 tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc
     5401 ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa
     5461 gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag
     5521 aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc
     5581 aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag
     5641 gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg
     5701 ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac
     5761 cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc
     5821 aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg
     5881 aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc
     5941 aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag
     6001 tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag
     6061 cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac
     6121 cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac
     6181 tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat
     6241 ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg
     6301 gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg
     6361 aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac
     6421 accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc
     6481 aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg
     6541 atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag
     6601 ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc
     6661 atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc
     6721 gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac
     6781 agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg
     6841 aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg
     6901 cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac
     6961 gatgtggacc acatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg
     7021 ctgaccagaa gcgacaaggc ccggggcaag agcgacaacg tgccctccga agaggtcgtg
     7081 aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag
     7141 ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc
     7201 atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac
     7261 tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc
     7321 accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc
     7381 gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc
     7441 ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac
     7501 gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac
     7561 ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag
     7621 atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag
     7681 ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa
     7741 aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc
     7801 gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc
     7861 cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa
     7921 ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag
     7981 aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc
     8041 aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct
     8101 gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg
     8161 tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag
     8221 ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc
     8281 tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag
     8341 caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc
     8401 aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac
     8461 accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac
     8521 gagacacgga tcgacctgtc tcagctggga ggcgacagcg ctggaggagg tggaagcgga
     8581 ggaggaggaa gcggaggagg aggtagcgga cctaagaaaa agaggaaggt ggcggccgct
     8641 ggatccagcg agctgattaa ggagaacatg cacatgaagc tctacatgga gggcaccgtg
     8701 gacaaccatc acttcaagtg cacatccgag ggcgaaggca agccctacga gggcacccag
     8761 accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga catcctggct
     8821 actagcttcc tctacggcag caagaccttc atcaaccaca cccagggcat ccccgacttc
     8881 ttcaagcagt ccttccctga gggcttcaca tgggagagag tcaccacata cgaagacggg
     8941 ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat ctacaacgtc
     9001 aagatcagag gggtgaactt cacatccaac ggccctgtga tgcagaagaa aacactcggc
     9061 tgggaggcct tcaccgaaac gctgtacccc gctgacggcg gcctggaagg cagaaacgac
     9121 atggccctga agctcgtggg cgggagccat ctgatcgcaa acatcaagac cacatataga
     9181 tccaagaaac ccgctaagaa cctcaagatg cctggcgtct actatgtgga ctacagactg
     9241 gaaagaatca aggaggccaa caacgagacc tacgtcgagc agcacgaggt ggcagtggcc
     9301 agatactgcg acctccctag caaactgggg cacaagctta atggcgcgcc aggttccaac
     9361 aattcccagg gaagagtgac cttcgaggat gtcactgtga acttcaccca gggggagtgg
     9421 cagcggctga atcccgaaca gagaaacttg tacagggatg tgatgctgga gaattacagc
     9481 aaccttgtct ctgtgggaca aggggaaacc accaaacccg atgtgatctt gaggttggaa
     9541 caaggaaagg agccatggtt ggaggaagag gaagtgctgg gaagtggccg tgcagaaaaa
     9601 aatggggaca ttggagggca gatttggaag ccaaaggatg tgaaagagag tctctaaaat
     9661 tcgatatcaa gcttatcggt aatcaacctc tggattacaa aatttgtgaa agattgactg
     9721 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     9781 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     9841 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     9901 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     9961 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
    10021 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
    10081 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
    10141 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
    10201 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
    10261 cctccccgca tcgataccgt cgacctcgag acctagaaaa acatggagca atcacaagta
    10321 gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg
    10381 tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag
    10441 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
    10501 gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct gattggcaga
    10561 actacacacc agggccaggg atcagatatc cactgacctt tggatggtgc tacaagctag
    10621 taccagttga gcaagagaag gtagaagaag ccaatgaagg agagaacacc cgcttgttac
    10681 accctgtgag cctgcatggg atggatgacc cggagagaga agtattagag tggaggtttg
    10741 acagccgcct agcatttcat cacatggccc gagagctgca tccggactgt actgggtctc
    10801 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
    10861 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
    10921 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagggccc
    10981 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
    11041 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
    11101 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
    11161 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
    11221 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
    11281 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
    11341 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
    11401 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
    11461 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
    11521 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
    11581 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
    11641 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
    11701 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
    11761 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
    11821 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
    11881 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
    11941 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
    12001 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
    12061 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
    12121 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc
    12181 cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct
    12241 cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac
    12301 cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg
    12361 ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg
    12421 ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc
    12481 cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt
    12541 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
    12601 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
    12661 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
    12721 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
    12781 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
    12841 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
    12901 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
    12961 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
    13021 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
    13081 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
    13141 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
    13201 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
    13261 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
    13321 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
    13381 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
    13441 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
    13501 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
    13561 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
    13621 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
    13681 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
    13741 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
    13801 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
    13861 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
    13921 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
    13981 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
    14041 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
    14101 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
    14161 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
    14221 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
    14281 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
    14341 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
    14401 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
    14461 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
    14521 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
    14581 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
    14641 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
    14701 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
    14761 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
    14821 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
    14881 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    14941 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgac
//
