LOCUS       sequence-234995-       14215 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral construct for inducible expression of dCas9-VP192 
            transcriptional activator and EGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14215)
  TITLE     Otonkoski plasmids 2018
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 14215)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: FU_tetO_dCas9-VP192-T2A-EGFP
FEATURES             Location/Qualifiers
     source          1..14215
                     /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"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    join(2610..2628,2629..2651,2652..2670,2671..2693,2694..2712,
                     2713..2735,2736..2754,2755..2777,2778..2796,2797..2819,
                     2820..2838,2839..2861,2862..2880)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2913..2951
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2948..2972
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     3051..3070
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3051..3069
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3125..3151
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3155..3175
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3185..3208,3209..3211,3212..5767,5768..5770,
                     5771..7285)
                     /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
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(3185..3208,3209..3211,3212..5698,5699..5701,
                     5702..7285)
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7289..7309
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7346..7456
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     CDS             7508..7618
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     CDS             7670..7780
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     CDS             7832..7942
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     CDS             7982..8035
                     /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             join(8042..8044,8045..8047,8048..8758)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(8042..8044,8045..8047,8048..8758)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8348..8367)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8695..8716
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             8789..8830
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             8840..8857
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    8907..9495
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8960..8980)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9378..9389)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9498..9514)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9499..9515)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10020..10200
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10226..10243)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10232..10456
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10502..10930
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10589..10608)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10799..10820
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10939..10959)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10944..11273
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11124..11259
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11186..11205
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11321..11368
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11387..11761
                     /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    11891..12012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11928..11947)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11982..12001
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12061..12077)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12061..12077)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12074..12096)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12085..12101
                     /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(12109..12115,12116..12133,12134..12139))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12154..12175
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12292..12309)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12463..13051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12543..12562)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13222..14013,14014..14082))
                     /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     13845..13864
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14083..14187)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttatcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc
     2641 gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat
     2701 cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg
     2761 aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact
     2821 ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga
     2881 aaagtgaaag tcgagctcgg tacccgggtc gaggtaggcg tgtacggtgg gaggcctata
     2941 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
     3001 acctccatag aagacaccgg gaccgatcca gcctccgcgg ccccgaattg taatacgact
     3061 cactataggg agacccaagc tggctaggta agcttgatca acaagtttgt acggtgccac
     3121 catgtaccca tacgatgttc cagattacgc ttcgccgaag aaaaagcgca aggtcgaagc
     3181 gtccgacaag aagtacagca tcggcctggc catcggcacc aactctgtgg gctgggccgt
     3241 gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg
     3301 gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga
     3361 ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg
     3421 ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag
     3481 actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg
     3541 caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa
     3601 gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca
     3661 catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga
     3721 cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc
     3781 catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag
     3841 acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggcaa
     3901 cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga
     3961 ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc
     4021 ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat
     4081 cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc
     4141 tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg
     4201 gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc
     4261 cggctacatt gacggcggag ccagccagga agagttctac aagttcatca agcccatcct
     4321 ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg
     4381 gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca
     4441 cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat
     4501 cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag
     4561 cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga
     4621 agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa
     4681 gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt
     4741 gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct
     4801 gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac
     4861 cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat
     4921 ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat
     4981 tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt
     5041 gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc
     5101 ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg
     5161 caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct
     5221 ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga
     5281 cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct
     5341 gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac
     5401 agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt
     5461 gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag
     5521 aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc
     5581 cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg
     5641 ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg atgtcgacgc
     5701 catcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag
     5761 cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa
     5821 gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct
     5881 gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca
     5941 gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa
     6001 cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc
     6061 caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa
     6121 ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa
     6181 gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa
     6241 gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag
     6301 caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg
     6361 gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt
     6421 tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt
     6481 gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat
     6541 cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc
     6601 ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt
     6661 gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga
     6721 ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa
     6781 gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact
     6841 gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag
     6901 ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga
     6961 acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt
     7021 gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa
     7081 gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc
     7141 ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa
     7201 agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat
     7261 cgacctgtct cagctgggag gcgacagccc caagaagaag agaaaggtgg aggccagcct
     7321 cgagtgcgaa agcggcgggc cggccgacgc ccttgacgat tttgacttag acatgctccc
     7381 agccgatgcc cttgacgact ttgaccttga tatgctgcct gctgacgctc ttgacgattt
     7441 tgaccttgac atgctccccg ggtcactacc cagcgctagc gtcgagtgcg aaagcggcgg
     7501 gccggccgac gcccttgacg attttgactt agacatgctc ccagccgatg cccttgacga
     7561 ctttgacctt gatatgctgc ctgctgacgc tcttgacgat tttgaccttg acatgctccc
     7621 cgggtcacta cccagcgcta gcgtcgagtg cgaaagcggc gggccggccg acgcccttga
     7681 cgattttgac ttagacatgc tcccagccga tgcccttgac gactttgacc ttgatatgct
     7741 gcctgctgac gctcttgacg attttgacct tgacatgctc cccgggtcac tacccagcgc
     7801 tagcgtcgag tgcgaaagcg gcgggccggc cgacgccctt gacgattttg acttagacat
     7861 gctcccagcc gatgcccttg acgactttga ccttgatatg ctgcctgctg acgctcttga
     7921 cgattttgac cttgacatgc tccccgggtc actacccagc gctagcgtcg acggctccgg
     7981 agagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccaatcga
     8041 tatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     8101 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     8161 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     8221 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     8281 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     8341 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     8401 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     8461 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     8521 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     8581 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     8641 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     8701 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaagt
     8761 ggttgatcta gagggcccgc ggttcgaagg taagcctatc cctaaccctc tcctcggtct
     8821 cgattctacg cgtaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     8881 ctcaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     8941 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     9001 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     9061 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     9121 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     9181 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     9241 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     9301 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     9361 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     9421 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     9481 gggccgcctc cccgcatcga taccgtcgac ctcgagacct agaaaaacat ggagcaatca
     9541 caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg
     9601 aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag
     9661 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     9721 aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt
     9781 ggcagaacta cacaccaggg ccagggatca gatatccact gacctttgga tggtgctaca
     9841 agctagtacc agttgagcaa gagaaggtag aagaagccaa tgaaggagag aacacccgct
     9901 tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagta ttagagtgga
     9961 ggtttgacag ccgcctagca tttcatcaca tggcccgaga gctgcatccg gactgtactg
    10021 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
    10081 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
    10141 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
    10201 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
    10261 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
    10321 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
    10381 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
    10441 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
    10501 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
    10561 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
    10621 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
    10681 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
    10741 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
    10801 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
    10861 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
    10921 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
    10981 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
    11041 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
    11101 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
    11161 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
    11221 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
    11281 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
    11341 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac
    11401 cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga
    11461 ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga
    11521 cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg
    11581 ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa
    11641 cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga
    11701 gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg
    11761 acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
    11821 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
    11881 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
    11941 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
    12001 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
    12061 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
    12121 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
    12181 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
    12241 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
    12301 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
    12361 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
    12421 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
    12481 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
    12541 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
    12601 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
    12661 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
    12721 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
    12781 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
    12841 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
    12901 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
    12961 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
    13021 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
    13081 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
    13141 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
    13201 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
    13261 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
    13321 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
    13381 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
    13441 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
    13501 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
    13561 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
    13621 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
    13681 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
    13741 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
    13801 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
    13861 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
    13921 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
    13981 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
    14041 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
    14101 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
    14161 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgac
//
