LOCUS       sequence-496078-       14815 bp    DNA     circular SYN 02-JAN-2026
DEFINITION  Lentiviral expression of dCas9-VP64 with H2B-GFP reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14815)
  TITLE     Brock Lab Plasmids 2025
REFERENCE   2  (bases 1 to 14815)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 2, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: lenti-EF1a-dCas9-VP64-H2B-GFP
FEATURES             Location/Qualifiers
     source          1..14815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      333..342
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(339..341,342..344,345..1058)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(339..341,342..344,345..1055)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(645..664)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     992..1013
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1084..1672
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1137..1157)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1555..1566)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1675..1691)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1676..1692)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2197..2377
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(2403..2420)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2409..2633
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2679..3107
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2766..2785)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2976..2997
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3116..3136)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3121..3450
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3301..3436
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3363..3382
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3498..3545
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3564..3938
                     /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    4068..4189
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4105..4124)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4159..4178
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4238..4254)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4238..4254)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4251..4273)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4262..4278
                     /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(4286..4292,4293..4310,4311..4316))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4331..4352
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4469..4486)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4640..5228)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4720..4739)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5399..6190,6191..6259))
                     /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     6022..6041
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6260..6364)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6439..6458)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6630..7009
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7011..7209
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7164..7184
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             7227..7407
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7454..7579
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8072..8305
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8490..8534
                     /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    8832..8949
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        9079..10257
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          9310..10248
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     10205..10225
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             10273..10320
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(10321..10344,10345..10347,10348..12903,12904..12906,
                     12907..14421)
                     /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             14476..14496
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             14521..14670
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             14692..14745
                     /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(14758..14815,1..320)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
ORIGIN
        1 ctaaggcgca gaagaaaggc ggcaagaagc gcaagcgcag ccgcaaggag agctattcca
       61 tctatgtgta caaggttctg aagcaggtcc accctgacac cggcatttcg tccaaggcca
      121 tgggcatcat gaattcgttt gtgaacgaca ttttcgagcg catcgcaggt gaggcttccc
      181 gcctggcgca ttacaacaag cgctcgacca tcacctccag ggagatccag acggccgtgc
      241 gcctgctgct gcctggggag ttggccaagc acgccgtgtc cgagggtact aaggccatca
      301 ccaagtacac cagcgctaag gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc
      361 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      421 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
      481 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
      541 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      601 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      661 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
      721 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
      781 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
      841 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
      901 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
      961 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1021 ccgggatcac tctcggcatg gacgagctgt acaagtaatt cgaacgatat caagcttatc
     1081 ggtaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     1141 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     1201 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     1261 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     1321 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     1381 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     1441 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     1501 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     1561 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     1621 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     1681 cgtcgacctc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     1741 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca
     1801 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     1861 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat
     1921 ctgtggatct accacacaca aggctacttc cctgattggc agaactacac accagggcca
     1981 gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag
     2041 aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat
     2101 gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt
     2161 catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc
     2221 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     2281 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     2341 ctcagaccct tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc
     2401 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2461 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2521 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2581 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     2641 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     2701 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2761 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2821 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2881 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2941 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3001 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3061 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3121 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3181 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3241 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3301 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3361 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     3421 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     3481 ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat
     3541 acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg
     3601 cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact
     3661 tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg
     3721 tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg
     3781 agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg
     3841 ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg
     3901 gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt
     3961 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     4021 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     4081 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4141 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     4201 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4261 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4321 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4381 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4441 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4501 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4561 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4621 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4681 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4741 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4801 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4861 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4921 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4981 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5041 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5101 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5161 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5221 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5281 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5341 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5401 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5461 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5521 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5581 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5641 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5701 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5761 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5821 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5881 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5941 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6001 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6061 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6121 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6181 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6241 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6301 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6361 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     6421 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     6481 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     6541 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     6601 tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
     6661 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     6721 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     6781 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     6841 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     6901 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     6961 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     7021 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     7081 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     7141 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     7201 tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg
     7261 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     7321 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     7381 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
     7441 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
     7501 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
     7561 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
     7621 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     7681 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     7741 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     7801 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     7861 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     7921 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     7981 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     8041 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     8101 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     8161 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     8221 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     8281 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     8341 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     8401 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     8461 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     8521 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     8581 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     8641 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     8701 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
     8761 ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca aatggcagta
     8821 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     8881 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     8941 caaaattttc gggtttatta cagggacagc agagatccag tttggttaat tagctagctg
     9001 caaagatgga taaagtttta aacagagagg aatctttgca gctaatggac cttctaggtc
     9061 ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca
     9121 gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc
     9181 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
     9241 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
     9301 agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc
     9361 ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg tgattcttga tcccgagctt
     9421 cgggttggaa gtgggtggga gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt
     9481 gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt
     9541 cgcgcctgtc tcgctgcttt cgataagtct ctagccattt aaaatttttg atgacctgct
     9601 gcgacgcttt ttttctggca agatagtctt gtaaatgcgg gccaagatct gcacactggt
     9661 atttcggttt ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg
     9721 gcgaggcggg gcctgcgagc gcggccaccg agaatcggac gggggtagtc tcaagctggc
     9781 cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg
     9841 ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca
     9901 gggagctcaa aatggaggac gcggcgctcg ggagagcggg cgggtgagtc acccacacaa
     9961 aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg actccacgga gtaccgggcg
    10021 ccgtccaggc acctcgatta gttctcgagc ttttggagta cgtcgtcttt aggttggggg
    10081 gaggggtttt atgcgatgga gtttccccac actgagtggg tggagactga agttaggcca
    10141 gcttggcact tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg atcttggttc
    10201 attctcaagc ctcagacagt ggttcaaagt ttttttcttc catttcaggt gtcgtgacgt
    10261 acggccacca tgaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag
    10321 gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg ggccgtgatc
    10381 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
    10441 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
    10501 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
    10561 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
    10621 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
    10681 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
    10741 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
    10801 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
    10861 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
    10921 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
    10981 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggcaacctg
    11041 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
    11101 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
    11161 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
    11221 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
    11281 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
    11341 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
    11401 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
    11461 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
    11521 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
    11581 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
    11641 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
    11701 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
    11761 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
    11821 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
    11881 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
    11941 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
    12001 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
    12061 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
    12121 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
    12181 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
    12241 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
    12301 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
    12361 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
    12421 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
    12481 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
    12541 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
    12601 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
    12661 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
    12721 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
    12781 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccacatc
    12841 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac
    12901 aaggcccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
    12961 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
    13021 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
    13081 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
    13141 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
    13201 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
    13261 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
    13321 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
    13381 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
    13441 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
    13501 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc
    13561 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
    13621 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc
    13681 agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat
    13741 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
    13801 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
    13861 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
    13921 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag
    13981 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
    14041 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
    14101 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
    14161 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc
    14221 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
    14281 gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag
    14341 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
    14401 ctgtctcagc tgggaggcga cagcgctgga ggaggtggaa gcggaggagg aggaagcgga
    14461 ggaggaggta gcggacctaa gaaaaagagg aaggtggcgg ccgctggatc cggacgggct
    14521 gacgcattgg acgattttga tctggatatg ctgggaagtg acgccctcga tgattttgac
    14581 cttgacatgc ttggttcgga tgcccttgat gactttgacc tcgacatgct cggcagtgac
    14641 gcccttgatg atttcgacct ggacatgctg attaactgta caggttcagg ggagggaaga
    14701 ggttccctgc tcacatgcgg agacgtagaa gagaatcccg gacccatgac cttcgaaatg
    14761 ccagagccag cgaagtctgc tcccgccccg aaaaagggct ccaagaaggc ggtga
//
