LOCUS       sequence-483039-       15841 bp    DNA     circular SYN 12-AUG-2025
DEFINITION  3rd generation lenti vector encoding dCas9 (s. Pyogenes) fused with 
            the VP64-p65-Rta(VPR) and a 2A GFP fluorescence marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15841)
  TITLE     Bowles lab plasmids 2025
REFERENCE   2  (bases 1 to 15841)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 12, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: lenti-EF1a-dCas9-VPR-eGFP
FEATURES             Location/Qualifiers
     source          1..15841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        130..1308
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          361..1299
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1256..1276
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1333..1342
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1339..1365,1366..1368,1369..3852,3853..3858,3859..3924,
                     3925..3927,3928..5442)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5455..5475
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    5512..5536
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5740..5760
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6190..6546
                     /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             6565..7134
                     /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             7150..7206
                     /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             join(7219..7221,7222..7935)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(7219..7221,7222..7932)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(7261..7282)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7522..7541)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7869..7890
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    7976..8564
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8029..8049)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8447..8458)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8567..8583)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8568..8584)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9089..9269
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9295..9312)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9301..9525
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9571..9999
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9658..9677)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9868..9889
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10008..10028)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10013..10342
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10193..10328
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10255..10274
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10390..10437
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10456..10830
                     /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    10960..11081
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10997..11016)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11051..11070
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11130..11146)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11130..11146)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11143..11165)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11154..11170
                     /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(11178..11184,11185..11202,11203..11208))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11223..11244
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11361..11378)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11532..12120)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11612..11631)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12291..13082,13083..13151))
                     /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     12914..12933
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13152..13256)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(13331..13350)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        13522..13901
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        13903..14101
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     14056..14076
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             14119..14299
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    14346..14471
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    14964..15197
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             15382..15426
                     /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    15724..15841
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cgggtttatt acagggacag cagagatcca gtttggttaa ttagctagct gcaaagatgg
       61 ataaagtttt aaacagagag gaatctttgc agctaatgga ccttctaggt cttgaaagga
      121 gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
      181 aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac
      241 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
      301 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
      361 gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg
      421 ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga
      481 agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt
      541 gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt
      601 ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt
      661 tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt
      721 tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg
      781 ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct
      841 ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg
      901 tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca
      961 aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg
     1021 gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg
     1081 cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt
     1141 tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac
     1201 ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag
     1261 cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgacg tacgctagag
     1321 gatcgaaccc ttgccaccat ggacaagaag tactccattg ggctcgctat cggcacaaac
     1381 agcgtcggct gggccgtcat tacggacgag tacaaggtgc cgagcaaaaa attcaaagtt
     1441 ctgggcaata ccgatcgcca cagcataaag aagaacctca ttggcgccct cctgttcgac
     1501 tccggggaga cggccgaagc cacgcggctc aaaagaacag cacggcgcag atatacccgc
     1561 agaaagaatc ggatctgcta cctgcaggag atctttagta atgagatggc taaggtggat
     1621 gactctttct tccataggct ggaggagtcc tttttggtgg aggaggataa aaagcacgag
     1681 cgccacccaa tctttggcaa tatcgtggac gaggtggcgt accatgaaaa gtacccaacc
     1741 atatatcatc tgaggaagaa gcttgtagac agtactgata aggctgactt gcggttgatc
     1801 tatctcgcgc tggcgcatat gatcaaattt cggggacact tcctcatcga gggggacctg
     1861 aacccagaca acagcgatgt cgacaaactc tttatccaac tggttcagac ttacaatcag
     1921 cttttcgaag agaacccgat caacgcatcc ggagttgacg ccaaagcaat cctgagcgct
     1981 aggctgtcca aatcccggcg gctcgaaaac ctcatcgcac agctccctgg ggagaagaag
     2041 aacggcctgt ttggtaatct tatcgccctg tcactcgggc tgacccccaa ctttaaatct
     2101 aacttcgacc tggccgaaga tgccaagctt caactgagca aagacaccta cgatgatgat
     2161 ctcgacaatc tgctggccca gatcggcgac cagtacgcag accttttttt ggcggcaaag
     2221 aacctgtcag acgccattct gctgagtgat attctgcgag tgaacacgga gatcaccaaa
     2281 gctccgctga gcgctagtat gatcaagcgc tatgatgagc accaccaaga cttgactttg
     2341 ctgaaggccc ttgtcagaca gcaactgcct gagaagtaca aggaaatttt cttcgatcag
     2401 tctaaaaatg gctacgccgg atacattgac ggcggagcaa gccaggagga attttacaaa
     2461 tttattaagc ccatcttgga aaaaatggac ggcaccgagg agctgctggt aaagcttaac
     2521 agagaagatc tgttgcgcaa acagcgcact ttcgacaatg gaagcatccc ccaccagatt
     2581 cacctgggcg aactgcacgc tatcctcagg cggcaagagg atttctaccc ctttttgaaa
     2641 gataacaggg aaaagattga gaaaatcctc acatttcgga taccctacta tgtaggcccc
     2701 ctcgcccggg gaaattccag attcgcgtgg atgactcgca aatcagaaga gaccatcact
     2761 ccctggaact tcgaggaagt cgtggataag ggggcctctg cccagtcctt catcgaaagg
     2821 atgactaact ttgataaaaa tctgcctaac gaaaaggtgc ttcctaaaca ctctctgctg
     2881 tacgagtact tcacagttta taacgagctc accaaggtca aatacgtcac agaagggatg
     2941 agaaagccag cattcctgtc tggagagcag aagaaagcta tcgtggacct cctcttcaag
     3001 acgaaccgga aagttaccgt gaaacagctc aaagaagact atttcaaaaa gattgaatgt
     3061 ttcgactctg ttgaaatcag cggagtggag gatcgcttca acgcatccct gggaacgtat
     3121 cacgatctcc tgaaaatcat taaagacaag gacttcctgg acaatgagga gaacgaggac
     3181 attcttgagg acattgtcct cacccttacg ttgtttgaag atagggagat gattgaagaa
     3241 cgcttgaaaa cttacgctca tctcttcgac gacaaagtca tgaaacagct caagaggcgc
     3301 cgatatacag gatgggggcg gctgtcaaga aaactgatca atgggatccg agacaagcag
     3361 agtggaaaga caatcctgga ttttcttaag tccgatggat ttgccaaccg gaacttcatg
     3421 cagttgatcc atgatgactc tctcaccttt aaggaggaca tccagaaagc acaagtttct
     3481 ggccaggggg acagtcttca cgagcacatc gctaatcttg caggtagccc agctatcaaa
     3541 aagggaatac tgcagaccgt taaggtcgtg gatgaactcg tcaaagtaat gggaaggcat
     3601 aagcccgaga atatcgttat cgagatggcc cgagagaacc aaactaccca gaagggacag
     3661 aagaacagta gggaaaggat gaagaggatt gaagagggta taaaagaact ggggtcccaa
     3721 atccttaagg aacacccagt tgaaaacacc cagcttcaga atgagaagct ctacctgtac
     3781 tacctgcaga acggcaggga catgtacgtg gatcaggaac tggacatcaa tcggctctcc
     3841 gactacgacg tggctgctat cgtgccccag tcttttctca aagatgattc tattgataat
     3901 aaagtgttga caagatccga taaagctaga gggaagagtg ataacgtccc ctcagaagaa
     3961 gttgtcaaga aaatgaaaaa ttattggcgg cagctgctga acgccaaact gatcacacaa
     4021 cggaagttcg ataatctgac taaggctgaa cgaggtggcc tgtctgagtt ggataaagcc
     4081 ggcttcatca aaaggcagct tgttgagaca cgccagatca ccaagcacgt ggcccaaatt
     4141 ctcgattcac gcatgaacac caagtacgat gaaaatgaca aactgattcg agaggtgaaa
     4201 gttattactc tgaagtctaa gctggtctca gatttcagaa aggactttca gttttataag
     4261 gtgagagaga tcaacaatta ccaccatgcg catgatgcct acctgaatgc agtggtaggc
     4321 actgcactta tcaaaaaata tcccaagctt gaatctgaat ttgtttacgg agactataaa
     4381 gtgtacgatg ttaggaaaat gatcgcaaag tctgagcagg aaataggcaa ggccaccgct
     4441 aagtacttct tttacagcaa tattatgaat tttttcaaga ccgagattac actggccaat
     4501 ggagagattc ggaagcgacc acttatcgaa acaaacggag aaacaggaga aatcgtgtgg
     4561 gacaagggta gggatttcgc gacagtccgg aaggtcctgt ccatgccgca ggtgaacatc
     4621 gttaaaaaga ccgaagtaca gaccggaggc ttctccaagg aaagtatcct cccgaaaagg
     4681 aacagcgaca agctgatcgc acgcaaaaaa gattgggacc ccaagaaata cggcggattc
     4741 gattctccta cagtcgctta cagtgtactg gttgtggcca aagtggagaa agggaagtct
     4801 aaaaaactca aaagcgtcaa ggaactgctg ggcatcacaa tcatggagcg atcaagcttc
     4861 gaaaaaaacc ccatcgactt tctcgaggcg aaaggatata aagaggtcaa aaaagacctc
     4921 atcattaagc ttcccaagta ctctctcttt gagcttgaaa acggccggaa acgaatgctc
     4981 gctagtgcgg gcgagctgca gaaaggtaac gagctggcac tgccctctaa atacgttaat
     5041 ttcttgtatc tggccagcca ctatgaaaag ctcaaagggt ctcccgaaga taatgagcag
     5101 aagcagctgt tcgtggaaca acacaaacac taccttgatg agatcatcga gcaaataagc
     5161 gaattctcca aaagagtgat cctcgccgac gctaacctcg ataaggtgct ttctgcttac
     5221 aataagcaca gggataagcc catcagggag caggcagaaa acattatcca cttgtttact
     5281 ctgaccaact tgggcgcgcc tgcagccttc aagtacttcg acaccaccat agacagaaag
     5341 cggtacacct ctacaaagga ggtcctggac gccacactga ttcatcagtc aattacgggg
     5401 ctctatgaaa caagaatcga cctctctcag ctcggtggag acagcagggc tgaccccaag
     5461 aagaagagga aggtgtcgcc agggatccgt cgacttgacg cgttgatatc aacaagtttg
     5521 tacaaaaaag caggctacaa agaggccagc ggttccggac gggctgacgc attggacgat
     5581 tttgatctgg atatgctggg aagtgacgcc ctcgatgatt ttgaccttga catgcttggt
     5641 tcggatgccc ttgatgactt tgacctcgac atgctcggca gtgacgccct ttatgatttc
     5701 gacctggaca tgctgattaa ctctagaagt tccggatctc cgaaaaagaa acgcaaagtt
     5761 ggtagccagt acctgcccga caccgacgac cggcaccgga tcgaggaaaa gcggaagcgg
     5821 acctacgaga cattcaagag catcatgaag aagtccccct tcagcggccc caccgaccct
     5881 agacctccac ctagaagaat cgccgtgccc agcagatcca gcgccagcgt gccaaaacct
     5941 gccccccagc cttacccctt caccagcagc ctgagcacca tcaactacga cgagttccct
     6001 accatggtgt tccccagcgg ccagatctct caggcctctg ctctggctcc agcccctcct
     6061 caggtgctgc ctcaggctcc tgctcctgca ccagctccag ccatggtgtc tgcactggct
     6121 caggcaccag cacccgtgcc tgtgctggct cctggacctc cacaggctgt ggctccacca
     6181 gcccctaaac ctacacaggc cggcgagggc acactgtctg aagctctgct gcagctgcag
     6241 ttcgacgacg aggatctggg agccctgctg ggaaacagca ccgatcctgc cgtgttcacc
     6301 gacctggcca gcgtggacaa cagcgagttc cagcagctgc tgaaccaggg catccctgtg
     6361 gcccctcaca ccaccgagcc catgctgatg gaataccccg aggccatcac ccggctcgtg
     6421 acaggcgctc agaggcctcc tgatccagct cctgcccctc tgggagcacc aggcctgcct
     6481 aatggactgc tgtctggcga cgaggacttc agctctatcg ccgatatgga tttctcagcc
     6541 ttgctgggct ctggcagcgg cagccgggat tccagggaag ggatgttttt gccgaagcct
     6601 gaggccggct ccgctattag tgacgtgttt gagggccgcg aggtgtgcca gccaaaacga
     6661 atccggccat ttcatcctcc aggaagtcca tgggccaacc gcccactccc cgccagcctc
     6721 gcaccaacac caaccggtcc agtacatgag ccagtcgggt cactgacccc ggcaccagtc
     6781 cctcagccac tggatccagc gcccgcagtg actcccgagg ccagtcacct gttggaggat
     6841 cccgatgaag agacgagcca ggctgtcaaa gcccttcggg agatggccga tactgtgatt
     6901 ccccagaagg aagaggctgc aatctgtggc caaatggacc tttcccatcc gcccccaagg
     6961 ggccatctgg atgagctgac aaccacactt gagtccatga ccgaggatct gaacctggac
     7021 tcacccctga ccccggaatt gaacgagatt ctggatacct tcctgaacga cgagtgcctc
     7081 ttgcatgcca tgcatatcag cacaggactg tccatcttcg acacatctct ctttgaattc
     7141 ggatctggtg ccacgaattt ctcgctactc aagcaggccg gtgatgtcga ggaaaaccct
     7201 ggtcctatgg tgtctagagt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     7261 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     7321 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     7381 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     7441 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     7501 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     7561 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     7621 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     7681 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     7741 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     7801 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     7861 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     7921 gagctgtaca agtaaagcgg ccgcgactct agaattcgat atcaagctta tcgataatca
     7981 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     8041 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     8101 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     8161 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     8221 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     8281 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     8341 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     8401 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     8461 agctgacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     8521 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc
     8581 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
     8641 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
     8701 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     8761 ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
     8821 ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc cagggatcag
     8881 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga
     8941 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga
     9001 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat
     9061 ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg
     9121 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     9181 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     9241 cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga
     9301 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     9361 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     9421 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     9481 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     9541 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     9601 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     9661 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     9721 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     9781 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     9841 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     9901 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     9961 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
    10021 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
    10081 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
    10141 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
    10201 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
    10261 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
    10321 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
    10381 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
    10441 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
    10501 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
    10561 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
    10621 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
    10681 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
    10741 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
    10801 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
    10861 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
    10921 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
    10981 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
    11041 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
    11101 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
    11161 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
    11221 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
    11281 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
    11341 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
    11401 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
    11461 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
    11521 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
    11581 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
    11641 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
    11701 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
    11761 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
    11821 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
    11881 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
    11941 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
    12001 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
    12061 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
    12121 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
    12181 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
    12241 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
    12301 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
    12361 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
    12421 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
    12481 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
    12541 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
    12601 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
    12661 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
    12721 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
    12781 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
    12841 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
    12901 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
    12961 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
    13021 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
    13081 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
    13141 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
    13201 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
    13261 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
    13321 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
    13381 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
    13441 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
    13501 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
    13561 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
    13621 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
    13681 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
    13741 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
    13801 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
    13861 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
    13921 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
    13981 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
    14041 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
    14101 gcgcgttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
    14161 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
    14221 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
    14281 gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag
    14341 aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc
    14401 ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt
    14461 gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa
    14521 ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag
    14581 aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg
    14641 gacagctaca accatccctt cagacaggat cagaagaact tagatcatta tataatacag
    14701 tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag
    14761 acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc
    14821 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
    14881 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
    14941 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
    15001 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt
    15061 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
    15121 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
    15181 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct
    15241 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc
    15301 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa
    15361 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca
    15421 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg
    15481 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga
    15541 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc
    15601 gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag atccattcga
    15661 ttagtgaacg gatcggcact gcgtgcgcca attctgcaga caaatggcag tattcatcca
    15721 caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
    15781 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
    15841 t
//
