LOCUS       sequence-379870-       14547 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lenti vector encoding VP64-dCas9-VP64 with 2A 
            ZsGreen1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14547)
  TITLE     Miner Lab plasmids
REFERENCE   2  (bases 1 to 14547)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: Lenti-EF1alpha-VP64-dCas9-VP64-ZsG
FEATURES             Location/Qualifiers
     source          1..14547
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2687..3865
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2918..3856
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3813..3833
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3872..3881
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3881..4030
                     /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             4046..4093
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(4094..4117,4118..4120,4121..6676,6677..6679,
                     6680..8194)
                     /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             8249..8269
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8294..8443
                     /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             8465..8518
                     /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             8519..9214
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     misc_feature    9239..9827
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9292..9312)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9710..9721)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9830..9846)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9831..9847)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10352..10532
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10558..10575)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10564..10788
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10834..11262
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10921..10940)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11131..11152
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11271..11291)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11276..11605
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11456..11591
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11518..11537
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11653..11700
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11719..12093
                     /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    12223..12344
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12260..12279)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12314..12333
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12393..12409)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12393..12409)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12406..12428)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12417..12433
                     /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(12441..12447,12448..12465,12466..12471))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12486..12507
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12624..12641)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12795..13383)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12875..12894)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13554..14345,14346..14414))
                     /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     14177..14196
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14415..14519)
                     /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 tggttaatta gctagctgca aagatggata aagttttaaa cagagaggaa
     2641 tctttgcagc taatggacct tctaggtctt gaaaggagtg ggaattggct ccggtgcccg
     2701 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     2761 ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     2821 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     2881 cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg
     2941 cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct
     3001 gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc
     3061 ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg
     3121 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct
     3181 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt
     3241 aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg
     3301 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag
     3361 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc
     3421 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga
     3481 aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg
     3541 agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc
     3601 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt
     3661 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac
     3721 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt
     3781 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt
     3841 ttttcttcca tttcaggtgt cgtgacgtac ggccaccatg gacgcattgg acgattttga
     3901 tctggatatg ctgggaagtg acgccctcga tgattttgac cttgacatgc ttggttcgga
     3961 tgcccttgat gactttgacc tcgacatgct cggcagtgac gcccttgatg atttcgacct
     4021 ggacatgctg ggatccgctg gctccaaaag gccggcggcc acgaaaaagg ccggccaggc
     4081 aaaaaagaaa aaggacaaga agtacagcat cggcctggcc atcggcacca actctgtggg
     4141 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
     4201 caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga
     4261 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
     4321 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
     4381 cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc
     4441 catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca
     4501 cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc
     4561 cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga
     4621 caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga
     4681 ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag
     4741 caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct
     4801 gttcggcaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga
     4861 cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa
     4921 cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc
     4981 cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct
     5041 gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc
     5101 tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa
     5161 cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa
     5221 gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga
     5281 cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg
     5341 agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg
     5401 ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag
     5461 gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa
     5521 cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa
     5581 cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta
     5641 cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc
     5701 cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg
     5761 gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc
     5821 cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct
     5881 gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga
     5941 agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa
     6001 aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac
     6061 cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa
     6121 gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat
     6181 ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg
     6241 cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat
     6301 cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga
     6361 gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag
     6421 ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa
     6481 agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca
     6541 gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga
     6601 tgtggaccac atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct
     6661 gaccagaagc gacaaggccc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa
     6721 gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt
     6781 cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat
     6841 caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc
     6901 ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac
     6961 cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga
     7021 gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct
     7081 gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga
     7141 cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt
     7201 cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat
     7261 ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg
     7321 ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa
     7381 gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga
     7441 taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc
     7501 caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact
     7561 gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa
     7621 tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa
     7681 gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc
     7741 cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta
     7801 cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct
     7861 gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc
     7921 caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca
     7981 ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa
     8041 tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac
     8101 cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga
     8161 gacacggatc gacctgtctc agctgggagg cgacagcgct ggaggaggtg gaagcggagg
     8221 aggaggaagc ggaggaggag gtagcggacc taagaaaaag aggaaggtgg cggccgctgg
     8281 atccggacgg gctgacgcat tggacgattt tgatctggat atgctgggaa gtgacgccct
     8341 cgatgatttt gaccttgaca tgcttggttc ggatgccctt gatgactttg acctcgacat
     8401 gctcggcagt gacgcccttg atgatttcga cctggacatg ctgattaact gtacaggcag
     8461 tggagagggc agaggaagtc tgctaacatg cggtgacgtc gaggagaatc ctggcccaat
     8521 ggcccagtcc aagcacggcc tgaccaagga gatgaccatg aagtaccgca tggagggctg
     8581 cgtggacggc cacaagttcg tgatcaccgg cgagggcatc ggctacccct tcaagggcaa
     8641 gcaggccatc aacctgtgcg tggtggaggg cggccccttg cccttcgccg aggacatctt
     8701 gtccgccgcc ttcatgtacg gcaaccgcgt gttcaccgag tacccccagg acatcgtcga
     8761 ctacttcaag aactcctgcc ccgccggcta cacctgggac cgctccttcc tgttcgagga
     8821 cggcgccgtg tgcatctgca acgccgacat caccgtgagc gtggaggaga actgcatgta
     8881 ccacgagtcc aagttctacg gcgtgaactt ccccgccgac ggccccgtga tgaagaagat
     8941 gaccgacaac tgggagccct cctgcgagaa gatcatcccc gtgcccaagc agggcatctt
     9001 gaagggcgac gtgagcatgt acctgctgct gaaggacggt ggccgcttgc gctgccagtt
     9061 cgacaccgtg tacaaggcca agtccgtgcc ccgcaagatg cccgactggc acttcatcca
     9121 gcacaagctg acccgcgagg accgcagcga cgccaagaac cagaagtggc acctgaccga
     9181 gcacgccatc gcctccggct ccgccttgcc ctaagaattc gatatcaagc ttatcggtaa
     9241 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     9301 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     9361 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     9421 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     9481 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     9541 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     9601 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     9661 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     9721 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     9781 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg
     9841 acctcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc
     9901 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca
     9961 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
    10021 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg
    10081 gatctaccac acacaaggct acttccctga ttggcagaac tacacaccag ggccagggat
    10141 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc aagagaaggt
    10201 agaagaagcc aatgaaggag agaacacccg cttgttacac cctgtgagcc tgcatgggat
    10261 ggatgacccg gagagagaag tattagagtg gaggtttgac agccgcctag catttcatca
    10321 catggcccga gagctgcatc cggactgtac tgggtctctc tggttagacc agatctgagc
    10381 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
    10441 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
    10501 acccttttag tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct
    10561 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
    10621 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
    10681 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
    10741 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
    10801 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
    10861 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
    10921 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
    10981 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
    11041 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
    11101 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
    11161 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
    11221 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
    11281 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
    11341 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
    11401 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
    11461 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
    11521 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
    11581 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
    11641 tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac
    11701 aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc
    11761 gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg
    11821 gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag
    11881 gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg
    11941 tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg
    12001 accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac
    12061 tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc
    12121 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
    12181 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
    12241 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
    12301 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
    12361 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
    12421 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
    12481 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
    12541 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
    12601 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
    12661 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
    12721 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
    12781 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
    12841 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
    12901 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
    12961 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
    13021 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
    13081 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
    13141 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
    13201 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
    13261 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
    13321 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
    13381 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
    13441 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
    13501 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
    13561 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
    13621 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
    13681 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
    13741 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
    13801 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
    13861 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
    13921 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
    13981 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
    14041 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
    14101 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
    14161 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
    14221 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
    14281 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
    14341 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
    14401 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
    14461 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
    14521 acatttcccc gaaaagtgcc acctgac
//
