LOCUS       sequence-496079-       16206 bp    DNA     circular SYN 02-JAN-2026
DEFINITION  Lentiviral expression of dCas9-VPR with H2B-GFP reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16206)
  TITLE     Brock Lab Plasmids 2025
REFERENCE   2  (bases 1 to 16206)
  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-VPR-H2B-GFP
FEATURES             Location/Qualifiers
     source          1..16206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(36..1214)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          45..983
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(68..88)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    1344..1461
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(1759..1803)
                     /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    1988..2221
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2714..2839
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             2886..3066
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(3084..3282)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3109..3129)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3284..3663
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3835..3854
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3929..4033
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4034..4102,4103..4894)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4252..4271)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5065..5653
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5554..5573
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5807..5824
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5941..5962
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5977..5982,5983..6000,6001..6007)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6015..6031
                     /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)."
     primer_bind     6020..6042
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6039..6055
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6039..6055
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6104..6225
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6115..6134)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6169..6188
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6355..6729)
                     /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"
     promoter        complement(6748..6795)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(6843..7172)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6857..6992
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6911..6930)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7157..7177
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(7186..7614)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7296..7317)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7508..7527
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    7660..7884
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     7873..7890
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             7916..8096
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     8601..8617
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     8602..8618
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8621..9209
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             8727..8738
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     9136..9156
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(9235..9948,9949..9951,9952..9954))
                     /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             complement(join(9238..9948,9949..9951,9952..9954))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(9280..9301)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     9629..9648
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      9951..9960
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(9973..10350)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             complement(10363..10416)
                     /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             complement(10427..10996)
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     CDS             complement(11015..11371)
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             complement(11801..11821)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(11829..11978)
                     /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             complement(12003..12023)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(12078..13592,13593..13595,13596..16151,
                     16152..16154,16155..16178))
                     /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             complement(join(16179..16206,1..20))
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 ttcgtggccg ccggcctttt catggtggcc gtacgtcacg acacctgaaa tggaagaaaa
       61 aaactttgaa ccactgtctg aggcttgaga atgaaccaag atccaaactc aaaaagggca
      121 aattccaagg agaattacat caagtgccaa gctggcctaa cttcagtctc cacccactca
      181 gtgtggggaa actccatcgc ataaaacccc tccccccaac ctaaagacga cgtactccaa
      241 aagctcgaga actaatcgag gtgcctggac ggcgcccggt actccgtgga gtcacatgaa
      301 gcgacggctg aggacggaaa ggcccttttc ctttgtgtgg gtgactcacc cgcccgctct
      361 cccgagcgcc gcgtcctcca ttttgagctc cctgcagcag ggccgggaag cggccatctt
      421 tccgctcacg caactggtgc cgaccgggcc agccttgccg cccagggcgg ggcgatacac
      481 ggcggcgcga ggccaggcac cagagcaggc cggccagctt gagactaccc ccgtccgatt
      541 ctcggtggcc gcgctcgcag gccccgcctc gccgaacatg tgcgctggga cgcacgggcc
      601 ccgtcgccgc ccgcggcccc aaaaaccgaa ataccagtgt gcagatcttg gcccgcattt
      661 acaagactat cttgccagaa aaaaagcgtc gcagcaggtc atcaaaaatt ttaaatggct
      721 agagacttat cgaaagcagc gagacaggcg cgaaggtgcc accagattcg cacgcggcgg
      781 ccccagcgcc caggccaggc ctcaactcaa gcacgaggcg aaggggctcc ttaagcgcaa
      841 ggcctcgaac tctcccaccc acttccaacc cgaagctcgg gatcaagaat cacgtactgc
      901 agccaggtgg aagtaattca aggcacgcaa gggccataac ccgtaaagag gccaggcccg
      961 cgggaaccac acacggcact tacctgtgtt ctggcggcaa acccgttgcg aaaaagaacg
     1021 ttcacggcga ctactgcact tatatacggt tctcccccac cctcgggaaa aaggcggagc
     1081 cagtacacga catcactttc ccagtttacc ccgcgccacc ttctctaggc accggttcaa
     1141 ttgccgaccc ctccccccaa cttctcgggg actgtgggcg atgtgcgctc tgcccactga
     1201 cgggcaccgg agccaattcc cactcctttc aagacctaga aggtccatta gctgcaaaga
     1261 ttcctctctg tttaaaactt tatccatctt tgcagctagc taattaacca aactggatct
     1321 ctgctgtccc tgtaataaac ccgaaaattt tgaatttttg taatttgttt ttgtaattct
     1381 ttagtttgta tgtctgttgc tattatgtct actattcttt cccctgcact gtacccccca
     1441 atcccccctt ttcttttaaa attgtggatg aatactgcca tttgtctgca gaattggcgc
     1501 acgcagtgcc gatccgttca ctaatcgaat ggatctgtct ctgtctctct ctccaccttc
     1561 ttcttctatt ccttcgggcc tgtcgggtcc cctcggggtt gggaggtggg tctgaaacga
     1621 taatggtgaa tatccctgcc taactctatt cactatagaa agtacagcaa aaactattct
     1681 taaacctacc aagcctccta ctatcattat gaataatttt atataccaca gccaatttgt
     1741 tatgttaaac caattccaca aacttgccca tttatctaat tccaataatt cttgttcatt
     1801 cttttcttgc tggttttgcg attcttcaat taaggagtgt attaagcttg tgtaattgtt
     1861 aatttctctg tcccactcca tccaggtcgt gtgattccaa atctgttcca gagatttatt
     1921 actccaacta gcattccaag gcacagcagt ggtgcaaatg agttttccag agcaacccca
     1981 aatccccagg agctgttgat cctttaggta tctttccaca gccaggattc ttgcctggag
     2041 ctgcttgatg ccccagactg tgagttgcaa cagatgctgt tgcgcctcaa tagccctcag
     2101 caaattgttc tgctgctgca ctataccaga caataattgt ctggcctgta ccgtcagcgt
     2161 cattgacgct gcgcccatag tgcttcctgc tgctcccaag aacccaagga acaaagctcc
     2221 tattcccact gctctttttt ctctctgcac cactcttctc tttgccttgg tgggtgctac
     2281 tcctaatggt tcaattttta ctactttata tttatataat tcacttctcc aattgtccct
     2341 catatctcct cctccaggtc tgaagatcag cggccgcttg ctgtgcggtg gtcttacttt
     2401 tgttttgctc ttcctctatc ttgtctaaag cttccttggt gtcttttatc tctatccttt
     2461 gatgcacaca atagagggtt gctactgtat tatataatga tctaagttct tctgatcctg
     2521 tctgaaggga tggttgtagc tgtcccagta tttgtctaca gccttctgat gtttctaaca
     2581 ggccaggatt aactgcgaat cgttctagct ccctgcttgc ccatactata tgttttaatt
     2641 tatatttttt ctttccccct ggccttaacc gaattttttc ccatcgcgat ctaattctcc
     2701 cccgcttaat actgacgctc tcgcacccat ctctctcctt ctagcctccg ctagtcaaaa
     2761 tttttggcgt actcaccagt cgccgcccct cgcctcttgc cgtgcgcgct tcagcaagcc
     2821 gagtcctgcg tcgagagagc tcctctggtt tccctttcgc tttcaagtcc ctgttcgggc
     2881 gccactgcta gagattttcc acactgacta aaagggtctg agggatctct agttaccaga
     2941 gtcacacaac agacgggcac acactacttg aagcactcaa ggcaagcttt attgaggctt
     3001 aagcagtggg ttccctagtt agccagagag ctcccaggct cagatctggt ctaaccagag
     3061 agacccagta caggcaaaac gcgctgctta tatagacctc ccaccgtaca cgcctaccgc
     3121 ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt
     3181 gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa
     3241 accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga
     3301 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
     3361 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca
     3421 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
     3481 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt
     3541 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat
     3601 atgggctatg aactaatgac cccgtaattg attactatta ataactagtc aataatcaat
     3661 gtcaacgcgt atatctggcc cgtacatcgc gaagcagcgc aaaacgccta accctaagca
     3721 gattcttcat gcaattgtcg gtcaagcctt gccttgttgt agcttaaatt ttgctcgcgc
     3781 actactcagc gacctccaac acacaagcag ggagcagata ctggcttaac tatgcggcat
     3841 cagagcagat tgtactgaga gtgcaccata ggggatcggg agatctcccg atccgtcgac
     3901 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3961 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4021 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4081 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4141 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4201 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4261 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4321 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4381 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4441 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4501 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4561 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4621 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4681 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4741 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4801 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4861 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4921 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4981 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5041 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5101 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5161 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5221 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5281 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5341 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5401 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5461 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5521 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5581 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5641 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5701 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5761 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     5821 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     5881 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     5941 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     6001 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     6061 ttacgccaag ctctagctag aggtcgacgg tatacagaca tgataagata cattgatgag
     6121 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     6181 gctattgctt tatttgtaac cattataagc tgcaataaac aagttggggt gggcgaagaa
     6241 ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc
     6301 gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtagca cgtgtcagtc
     6361 ctgctcctcg gccacgaagt gcacgcagtt gccggccggg tcgcgcaggg cgaactcccg
     6421 cccccacggc tgctcgccga tctcggtcat ggccggcccg gaggcgtccc ggaagttcgt
     6481 ggacacgacc tccgaccact cggcgtacag ctcgtccagg ccgcgcaccc acacccaggc
     6541 cagggtgttg tccggcacca cctggtcctg gaccgcgctg atgaacaggg tcacgtcgtc
     6601 ccggaccaca ccggcgaagt cgtcctccac gaagtcccgg gagaacccga gccggtcggt
     6661 ccagaactcg accgctccgg cgacgtcgcg cgcggtgagc accggaacgg cactggtcaa
     6721 cttggccatg gtttagttcc tcaccttgtc gtattatact atgccgatat actatgccga
     6781 tgattaattg tcaacacgtg ctgatcagat ccgaaaatgg atatacaagc tcccgggagc
     6841 tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag
     6901 gcagaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa
     6961 tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt
     7021 gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt
     7081 ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag
     7141 cctggggact ttccacaccc taactgacac acattccaca gaattaattc gcgttaaatt
     7201 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     7261 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     7321 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     7381 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     7441 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     7501 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     7561 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtggggat
     7621 accccctaga gccccagctg gttctttccg cctcagaagc catagagccc accgcatccc
     7681 cagcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc
     7741 ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa
     7801 ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag
     7861 atggctggca actagaaggc acagtcgagg ctgatcagcg ggtttaaacg ggccctgcta
     7921 gagattttcc acactgacta aaagggtctg agggatctct agttaccaga gtcacacaac
     7981 agacgggcac acactacttg aagcactcaa ggcaagcttt attgaggctt aagcagtggg
     8041 ttccctagtt agccagagag ctcccaggct cagatctggt ctaaccagag agacccagta
     8101 cagtccggat gcagctctcg ggccatgtga tgaaatgcta ggcggctgtc aaacctccac
     8161 tctaatactt ctctctccgg gtcatccatc ccatgcaggc tcacagggtg taacaagcgg
     8221 gtgttctctc cttcattggc ttcttctacc ttctcttgct caactggtac tagcttgtag
     8281 caccatccaa aggtcagtgg atatctgatc cctggccctg gtgtgtagtt ctgccaatca
     8341 gggaagtagc cttgtgtgtg gtagatccac agatcaagga tatcttgtct tcgttgggag
     8401 tgaattagcc cttccagtcc ccccttttct tttaaaaagt ggctaagatc tacagctgcc
     8461 ttgtaagtca ttggtcttaa aggtacctga ggtgtgactg gaaaacccac ctcctcctcc
     8521 tcttgtgctt ctagccaggc acaatcagca ttggtagctg ctgtattgct acttgtgatt
     8581 gctccatgtt tttctaggtc tcgaggtcga cggtatcgat gcggggaggc ggcccaaagg
     8641 gagatccgac tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc
     8701 aggccgcggg aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc
     8761 ccgcgcagaa tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc
     8821 gacaacacca cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg
     8881 caggcggcga tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg
     8941 agctgacagg tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac
     9001 accacgccac gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat
     9061 ttatacaagg aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt
     9121 aaagcagcgt atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt
     9181 tcacaaattt tgtaatccag aggttgatta ccgataagct tgatatcgtt cgaattactt
     9241 gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat
     9301 gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac tgggtgctca ggtagtggtt
     9361 gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag
     9421 ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga tgccgttctt
     9481 ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc
     9541 caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt
     9601 gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat
     9661 ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat
     9721 gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca cgagggtggg
     9781 ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgtaggt
     9841 ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag
     9901 ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatggtggc
     9961 gaccggtgga tccttagcgc tggtgtactt ggtgatggcc ttagtaccct cggacacggc
    10021 gtgcttggcc aactccccag gcagcagcag gcgcacggcc gtctggatct ccctggaggt
    10081 gatggtcgag cgcttgttgt aatgcgccag gcgggaagcc tcacctgcga tgcgctcgaa
    10141 aatgtcgttc acaaacgaat tcatgatgcc catggccttg gacgaaatgc cggtgtcagg
    10201 gtggacctgc ttcagaacct tgtacacata gatggaatag ctctccttgc ggctgcgctt
    10261 gcgcttcttg ccgcctttct tctgcgcctt agtcaccgcc ttcttggagc cctttttcgg
    10321 ggcgggagca gacttcgctg gctctggcat ttcgaaggtc atgggtccgg gattctcttc
    10381 tacgtctccg catgtgagca gggaacctct tccctcccct gaacctaaac agagatgtgt
    10441 cgaagatgga cagtcctgtg ctgatatgca tggcatgcaa gaggcactcg tcgttcagga
    10501 aggtatccag aatctcgttc aattccgggg tcaggggtga gtccaggttc agatcctcgg
    10561 tcatggactc aagtgtggtt gtcagctcat ccagatggcc ccttgggggc ggatgggaaa
    10621 ggtccatttg gccacagatt gcagcctctt ccttctgggg aatcacagta tcggccatct
    10681 cccgaagggc tttgacagcc tggctcgtct cttcatcggg atcctccaac aggtgactgg
    10741 cctcgggagt cactgcgggc gctggatcca gtggctgagg gactggtgcc ggggtcagtg
    10801 acccgactgg ctcatgtact ggaccggttg gtgttggtgc gaggctggcg gggagtgggc
    10861 ggttggccca tggacttcct ggaggatgaa atggccggat tcgttttggc tggcacacct
    10921 cgcggccctc aaacacgtca ctaatagcgg agccggcctc aggcttcggc aaaaacatcc
    10981 cttccctgga atcccggctg ccgctgccag agcccagcaa ggctgagaaa tccatatcgg
    11041 cgatagagct gaagtcctcg tcgccagaca gcagtccatt aggcaggcct ggtgctccca
    11101 gaggggcagg agctggatca ggaggcctct gagcgcctgt cacgagccgg gtgatggcct
    11161 cggggtattc catcagcatg ggctcggtgg tgtgaggggc cacagggatg ccctggttca
    11221 gcagctgctg gaactcgctg ttgtccacgc tggccaggtc ggtgaacacg gcaggatcgg
    11281 tgctgtttcc cagcagggct cccagatcct cgtcgtcgaa ctgcagctgc agcagagctt
    11341 cagacagtgt gccctcgccg gcctgtgtag gtttaggggc tggtggagcc acagcctgtg
    11401 gaggtccagg agccagcaca ggcacgggtg ctggtgcctg agccagtgca gacaccatgg
    11461 ctggagctgg tgcaggagca ggagcctgag gcagcacctg aggaggggct ggagccagag
    11521 cagaggcctg agagatctgg ccgctgggga acaccatggt agggaactcg tcgtagttga
    11581 tggtgctcag gctgctggtg aaggggtaag gctggggggc aggttttggc acgctggcgc
    11641 tggatctgct gggcacggcg attcttctag gtggaggtct agggtcggtg gggccgctga
    11701 agggggactt cttcatgatg ctcttgaatg tctcgtaggt ccgcttccgc ttttcctcga
    11761 tccggtgccg gtcgtcggtg tcgggcaggt actggctacc aactttgcgt ttctttttcg
    11821 gagttaatca gcatgtccag gtcgaaatca tcaagggcgt cactgccgag catgtcgagg
    11881 tcaaagtcat caagggcatc cgaaccaagc atgtcaaggt caaaatcatc gagggcgtca
    11941 cttcccagca tatccagatc aaaatcgtcc aatgcgtcag cccgtccgga tccagcggcc
    12001 gccaccttcc tctttttctt aggtccgcta cctcctcctc cgcttcctcc tcctccgctt
    12061 ccacctcctc cagcgctgtc gcctcccagc tgagacaggt cgatccgtgt ctcgtacagg
    12121 ccggtgatgc tctggtggat cagggtggcg tccagcacct ctttggtgct ggtgtacctc
    12181 ttccggtcga tggtggtgtc aaagtacttg aaggcggcag gggctcccag attggtcagg
    12241 gtaaacaggt ggatgatatt ctcggcctgc tctctgatgg gcttatcccg gtgcttgttg
    12301 taggcggaca gcactttgtc cagattagcg tcggccagga tcactctctt ggagaactcg
    12361 ctgatctgct cgatgatctc gtccaggtag tgcttgtgct gttccacaaa cagctgtttc
    12421 tgctcattat cctcggggga gcccttcagc ttctcatagt ggctggccag gtacaggaag
    12481 ttcacatatt tggagggcag ggccagttcg tttcccttct gcagttcgcc ggcagaggcc
    12541 agcattctct tccggccgtt ttccagctcg aacagggagt acttaggcag cttgatgatc
    12601 aggtcctttt tcacttcttt gtagcccttg gcttccagaa agtcgatggg attcttctcg
    12661 aagctgcttc tttccatgat ggtgatcccc agcagctctt tcacactctt cagtttcttg
    12721 gacttgccct tttccacttt ggccaccacc agcacagaat aggccacggt ggggctgtcg
    12781 aagccgccgt acttcttagg gtcccagtcc ttctttctgg cgatcagctt atcgctgttc
    12841 ctcttgggca ggatagactc tttgctgaag ccgcctgtct gcacctcggt ctttttcacg
    12901 atattcactt ggggcatgct cagcactttc cgcacggtgg caaaatcccg gcccttatcc
    12961 cacacgatct ccccggtttc gccgtttgtc tcgatcagag gccgcttccg gatctcgccg
    13021 ttggccaggg taatctcggt cttgaaaaag ttcatgatgt tgctgtagaa gaagtacttg
    13081 gcggtagcct tgccgatttc ctgctcgctc ttggcgatca tcttccgcac gtcgtacacc
    13141 ttgtagtcgc cgtacacgaa ctcgctttcc agcttagggt actttttgat cagggcggtt
    13201 cccacgacgg cgttcaggta ggcgtcgtgg gcgtggtggt agttgttgat ctcgcgcact
    13261 ttgtaaaact ggaaatcctt ccggaaatcg gacaccagct tggacttcag ggtgatcact
    13321 ttcacttccc ggatcagctt gtcattctcg tcgtacttag tgttcatccg ggagtccagg
    13381 atctgtgcca cgtgctttgt gatctgccgg gtttccacca gctgtctctt gatgaagccg
    13441 gccttatcca gttcgctcag gccgcctctc tcggccttgg tcagattgtc gaactttctc
    13501 tgggtaatca gcttggcgtt cagcagctgc cgccagtagt tcttcatctt cttcacgacc
    13561 tcttcggagg gcacgttgtc gctcttgccc cgggccttgt cgcttctggt cagcaccttg
    13621 ttgtcgatgg agtcgtcctt cagaaagctc tgaggcacga tgtggtccac atcgtagtcg
    13681 gacagccggt tgatgtccag ttcctggtcc acgtacatat cccgcccatt ctgcaggtag
    13741 tacaggtaca gcttctcgtt ctgcagctgg gtgttttcca cggggtgttc tttcaggatc
    13801 tggctgccca gctctttgat gccctcttcg atccgcttca ttctctcgcg gctgttcttc
    13861 tgtcccttct gggtggtctg gttctctctg gccatttcga tcacgatgtt ctcgggcttg
    13921 tgccggccca tcactttcac gagctcgtcc accaccttca ctgtctgcag gatgcccttc
    13981 ttaatggcgg ggctgccggc cagattggca atgtgctcgt gcaggctatc gccctggccg
    14041 gacacctggg ctttctggat gtcctcttta aaggtcaggc tgtcgtcgtg gatcagctgc
    14101 atgaagtttc tgttggcgaa gccgtcggac ttcaggaaat ccaggattgt cttgccggac
    14161 tgcttgtccc ggatgccgtt gatcagcttc cggctcagcc tgccccagcc ggtgtatctc
    14221 cgccgcttca gctgcttcat cactttgtcg tcgaacaggt gggcataggt tttcagccgt
    14281 tcctcgatca tctctctgtc ctcaaacagt gtcagggtca gcacgatatc ttccagaatg
    14341 tcctcgtttt cctcattgtc caggaagtcc ttgtccttga taattttcag cagatcgtgg
    14401 tatgtgccca gggaggcgtt gaaccgatct tccacgccgg agatttccac ggagtcgaag
    14461 cactcgattt tcttgaagta gtcctctttc agctgcttca cggtcacttt ccggttggtc
    14521 ttgaacagca ggtccacgat ggcctttttc tgctcgccgc tcaggaaggc gggctttctc
    14581 attccctcgg tcacgtattt cactttggtc agctcgttat acacggtgaa gtactcgtac
    14641 agcaggctgt gcttgggcag caccttctcg ttgggcaggt tcttatcgaa gttggtcatc
    14701 cgctcgatga agctctgggc ggaagcgccc ttgtccacca cttcctcgaa gttccagggg
    14761 gtgatggttt cctcgctctt tctggtcatc caggcgaatc tgctgtttcc cctggccaga
    14821 gggcccacgt agtaggggat gcggaaggtc aggatcttct cgatcttttc ccggttgtcc
    14881 ttcaggaatg ggtaaaaatc ttcctgccgc cgcagaatgg cgtgcagctc tcccaggtgg
    14941 atctggtggg ggatgctgcc gttgtcgaag gtccgctgct tccgcagcag gtcctctctg
    15001 ttcagcttca cgagcagttc ctcggtgccg tccatctttt ccaggatggg cttgatgaac
    15061 ttgtagaact cttcctggct ggctccgccg tcaatgtagc cggcgtagcc gttcttgctc
    15121 tggtcgaaga aaatctcttt gtacttctca ggcagctgct gccgcacgag agctttcagc
    15181 agggtcaggt cctggtggtg ctcgtcgtat ctcttgatca tagaggcgct caggggggcc
    15241 ttggtgatct cggtgttcac tctcaggatg tcgctcagca ggatggcgtc ggacaggttc
    15301 ttggcggcca gaaacaggtc ggcgtactgg tcgccgatct gggccagcag gttgtccagg
    15361 tcgtcgtcgt aggtgtcctt gctcagctgc agtttggcat cctcggccag gtcgaagttg
    15421 ctcttgaagt tgggggtcag gcccaggctc agggcaatca ggttgccgaa caggccattc
    15481 ttcttctcgc cgggcagctg ggcgatcaga ttttccagcc gtctgctctt gctcagtctg
    15541 gcagacagga tggccttggc gtccacgccg ctggcgttga tggggttttc ctcgaacagc
    15601 tggttgtagg tctgcaccag ctggatgaac agcttgtcca cgtcgctgtt gtcggggttc
    15661 aggtcgccct cgatcaggaa gtggccccgg aacttgatca tgtgggccag ggccagatag
    15721 atcagccgca ggtcggcctt gtcggtgctg tccaccagtt tctttctcag gtggtagatg
    15781 gtggggtact tctcgtggta ggccacctcg tccacgatgt tgccgaagat ggggtgccgc
    15841 tcgtgcttct tatcctcttc caccaggaag gactcttcca gtctgtggaa gaagctgtcg
    15901 tccaccttgg ccatctcgtt gctgaagatc tcttgcagat agcagatccg gttcttccgt
    15961 ctggtgtatc ttcttctggc ggttctcttc agccgggtgg cctcggctgt ttcgccgctg
    16021 tcgaacagca gggctccgat caggttcttc ttgatgctgt gccggtcggt gttgcccagc
    16081 accttgaatt tcttgctggg caccttgtac tcgtcggtga tcacggccca gcccacagag
    16141 ttggtgccga tggccaggcc gatgctgtac ttcttgtcct ttttcttttt tgcctggccg
    16201 gccttt
//
