LOCUS       sequence-379871-       15126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lenti vector encoding dCas9-VP64 with P2A MS2-VP64 
            and T2A ZsGreen1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15126)
  TITLE     Miner Lab plasmids
REFERENCE   2  (bases 1 to 15126)
  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-dCas9-VP64-MS2-VP64-ZsG
FEATURES             Location/Qualifiers
     source          1..15126
                     /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"
     CDS             3881..3928
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(3929..3952,3953..3955,3956..6511,6512..6514,
                     6515..8029)
                     /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             8084..8104
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8129..8278
                     /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             8300..8356
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             8357..8743
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2-N55K
                     /note="binds with high affinity to a specific stem-loop 
                     structure in the viral RNA (Lim et al., 1994)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     CDS             8798..8818
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8858..9007
                     /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             9044..9097
                     /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             9098..9793
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     misc_feature    9818..10406
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9871..9891)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10289..10300)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10409..10425)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10410..10426)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10931..11111
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11137..11154)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11143..11367
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11413..11841
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11500..11519)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(11850..11870)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11855..12184
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12035..12170
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12097..12116
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12232..12279
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12298..12672
                     /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    12802..12923
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12839..12858)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12893..12912
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12972..12988)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12972..12988)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12985..13007)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12996..13012
                     /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(13020..13026,13027..13044,13045..13050))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13065..13086
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13203..13220)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13374..13962)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13454..13473)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14133..14924,14925..14993))
                     /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     14756..14775
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14994..15098)
                     /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 aaaaggccgg cggccacgaa
     3901 aaaggccggc caggcaaaaa agaaaaagga caagaagtac agcatcggcc tggccatcgg
     3961 caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt
     4021 caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct
     4081 gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata
     4141 caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa
     4201 ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa
     4261 gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta
     4321 ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg
     4381 gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg
     4441 cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta
     4501 caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct
     4561 gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga
     4621 gaagaagaat ggcctgttcg gcaacctgat tgccctgagc ctgggcctga cccccaactt
     4681 caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga
     4741 cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc
     4801 cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat
     4861 caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct
     4921 gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt
     4981 cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt
     5041 ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa
     5101 gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca
     5161 ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt
     5221 cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt
     5281 gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac
     5341 catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat
     5401 cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag
     5461 cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga
     5521 gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct
     5581 gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat
     5641 cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg
     5701 cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa
     5761 cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat
     5821 cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa
     5881 gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga
     5941 caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa
     6001 cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca
     6061 ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc
     6121 cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg
     6181 ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa
     6241 gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg
     6301 cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta
     6361 cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg
     6421 gctgtccgac tacgatgtgg accacatcgt gcctcagagc tttctgaagg acgactccat
     6481 cgacaacaag gtgctgacca gaagcgacaa ggcccggggc aagagcgaca acgtgccctc
     6541 cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat
     6601 tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga
     6661 taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc
     6721 acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga
     6781 agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt
     6841 ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt
     6901 cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga
     6961 ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc
     7021 taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct
     7081 ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat
     7141 cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt
     7201 gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc
     7261 caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg
     7321 cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg
     7381 caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag
     7441 cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa
     7501 ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag
     7561 aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata
     7621 tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa
     7681 tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca
     7741 gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc
     7801 cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct
     7861 gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga
     7921 ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat
     7981 caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca gcgctggagg
     8041 aggtggaagc ggaggaggag gaagcggagg aggaggtagc ggacctaaga aaaagaggaa
     8101 ggtggcggcc gctggatccg gacgggctga cgcattggac gattttgatc tggatatgct
     8161 gggaagtgac gccctcgatg attttgacct tgacatgctt ggttcggatg cccttgatga
     8221 ctttgacctc gacatgctcg gcagtgacgc ccttgatgat ttcgacctgg acatgctgat
     8281 taactgtaca ggcagtggag caacaaactt ctctctgctg aaacaagccg gagatgtcga
     8341 agagaatcct ggaccggctt caaactttac tcagttcgtg ctcgtggaca atggtgggac
     8401 aggggatgtg acagtggctc cttctaattt cgctaatggg gtggcagagt ggatcagctc
     8461 caactcacgg agccaggcct acaaggtgac atgcagcgtc aggcagtcta gtgcccagaa
     8521 gagaaagtat accatcaagg tggaggtccc caaagtggct acccagacag tgggcggagt
     8581 cgaactgcct gtcgccgctt ggaggtccta cctgaacatg gagctcacta tcccaatttt
     8641 cgctaccaat tctgactgtg aactcatcgt gaaggcaatg caggggctcc tcaaagacgg
     8701 taatcctatc ccttccgcca tcgccgctaa ctcaggtatc tacagcgctg gaggaggtgg
     8761 aagcggagga ggaggaagcg gaggaggagg tagcggacct aagaaaaaga ggaaggtgaa
     8821 aaagaggaag gtggcggccg ctggatccgg acgggctgac gcattggacg attttgatct
     8881 ggatatgctg ggaagtgacg ccctcgatga ttttgacctt gacatgcttg gttcggatgc
     8941 ccttgatgac tttgacctcg acatgctcgg cagtgacgcc cttgatgatt tcgacctgga
     9001 catgctggga tccgctggct ccgctgctgg ttctggcgaa ttcgagggca gaggaagtct
     9061 gctaacatgc ggtgacgtcg aggagaatcc tggcccaatg gcccagtcca agcacggcct
     9121 gaccaaggag atgaccatga agtaccgcat ggagggctgc gtggacggcc acaagttcgt
     9181 gatcaccggc gagggcatcg gctacccctt caagggcaag caggccatca acctgtgcgt
     9241 ggtggagggc ggccccttgc ccttcgccga ggacatcttg tccgccgcct tcatgtacgg
     9301 caaccgcgtg ttcaccgagt acccccagga catcgtcgac tacttcaaga actcctgccc
     9361 cgccggctac acctgggacc gctccttcct gttcgaggac ggcgccgtgt gcatctgcaa
     9421 cgccgacatc accgtgagcg tggaggagaa ctgcatgtac cacgagtcca agttctacgg
     9481 cgtgaacttc cccgccgacg gccccgtgat gaagaagatg accgacaact gggagccctc
     9541 ctgcgagaag atcatccccg tgcccaagca gggcatcttg aagggcgacg tgagcatgta
     9601 cctgctgctg aaggacggtg gccgcttgcg ctgccagttc gacaccgtgt acaaggccaa
     9661 gtccgtgccc cgcaagatgc ccgactggca cttcatccag cacaagctga cccgcgagga
     9721 ccgcagcgac gccaagaacc agaagtggca cctgaccgag cacgccatcg cctccggctc
     9781 cgccttgccc taagaattcg atatcaagct tatcggtaat caacctctgg attacaaaat
     9841 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     9901 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     9961 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
    10021 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
    10081 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
    10141 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
    10201 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
    10261 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
    10321 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
    10381 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgagacc tagaaaaaca
    10441 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc
    10501 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac
    10561 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
    10621 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta
    10681 cttccctgat tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg
    10741 atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga
    10801 gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt
    10861 attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc
    10921 ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
    10981 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
    11041 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
    11101 aatctctagc agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
    11161 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
    11221 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
    11281 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
    11341 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
    11401 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
    11461 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
    11521 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
    11581 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
    11641 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
    11701 tctttaatag tggactcttg ttccaaactg taacaacact caaccctatc tcggtctatt
    11761 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
    11821 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
    11881 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
    11941 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
    12001 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
    12061 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
    12121 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
    12181 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
    12241 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
    12301 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag
    12361 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg
    12421 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac
    12481 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc
    12541 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg
    12601 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag
    12661 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
    12721 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
    12781 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
    12841 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
    12901 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
    12961 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
    13021 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
    13081 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
    13141 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
    13201 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
    13261 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
    13321 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
    13381 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
    13441 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
    13501 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
    13561 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
    13621 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
    13681 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
    13741 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
    13801 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
    13861 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
    13921 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
    13981 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
    14041 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
    14101 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
    14161 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
    14221 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
    14281 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
    14341 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
    14401 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
    14461 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
    14521 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
    14581 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
    14641 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
    14701 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
    14761 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
    14821 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
    14881 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
    14941 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
    15001 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
    15061 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
    15121 cctgac
//
