LOCUS       sequence-211518-       12425 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Lentiviral vector for constitutive expression of Cas9-VQR-P2A-GFP 
            (not codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12425)
  AUTHORS   Zafra MP, Schatoff EM, Katti A, Foronda M, Breinig M, Schweitzer AY,
            Simon A, Han T, Goswami S, Montgomery E, Thibado J, Kastenhuber ER, 
            Sanchez-Rivera FJ, Shi J, Vakoc CR, Lowe SW, Tschaharganeh DF, Dow 
            LE
  TITLE     Optimized base editors enable efficient editing in cells, organoids 
            and mice.
  JOURNAL   Nat Biotechnol. 2018 Jul 3. pii: nbt.4194. doi: 10.1038/nbt.4194.
  PUBMED    29969439
REFERENCE   2  (bases 1 to 12425)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2018 Jul 3. pii: nbt.4194. doi: 10.1038/nbt.4194."
COMMENT     Sequence Label: pLenti-VQR-P2A-GFP-PGK-Puro
FEATURES             Location/Qualifiers
     source          1..12425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             232..412
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1077..1310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1495..1539
                     /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    1723..1840
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1909..2120
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2140..2149
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2146..5547,5548..5550,5551..6147,6148..6150,6151..6153,
                     6154..6156,6157..6249)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6256..6276
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6283..6348
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6358..6414
                     /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(6415..6417,6418..6420,6421..7134)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6460..6481)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6721..6740)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7068..7089
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        7147..7646
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7182..7205)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7579..7598
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             7667..8266
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7667..7686)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8163..8183
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8282..8870
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8335..8355)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8753..8764)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8942..9175
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9253..9374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9290..9309)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9344..9363
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9414..9549
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9569..9588)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9570..9588)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9598..9615)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9598..9614)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9607..9629)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9756..10211
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9843..9862)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10053..10074
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10237..10341
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10342..10410,10411..11202)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(10560..10579)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11373..11961
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11862..11881
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12115..12132
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12249..12270
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12285..12290,12291..12308,12309..12315)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12323..12339
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     12328..12350
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12347..12363
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12347..12363
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12382..12402
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        12384..12402
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagggga cccagaccac aattttaaaa gaaaaggggg
     1741 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     1801 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     1861 cagagatcca ctttggcgcc ggctcgagtg gctccggtgc ccgtcagtgg gcagagcgca
     1921 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
     1981 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     2041 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     2101 ggtttgccgc cagaacacag gtgtcgtgac gcgggatccg ccaccatgga taaaaagtat
     2161 tctattggtt tagacatcgg cactaattcc gttggatggg ctgtcataac cgatgaatac
     2221 aaagtacctt caaagaaatt taaggtgttg gggaacacag accgtcattc gattaaaaag
     2281 aatcttatcg gtgccctcct attcgatagt ggcgaaacgg cagaggcgac tcgcctgaaa
     2341 cgaaccgctc ggagaaggta tacacgtcgc aagaaccgaa tatgttactt acaagaaatt
     2401 tttagcaatg agatggccaa agttgacgat tctttctttc accgtttgga agagtccttc
     2461 cttgtcgaag aggacaagaa acatgaacgg caccccatct ttggaaacat agtagatgag
     2521 gtggcatatc atgaaaagta cccaacgatt tatcacctca gaaaaaagct agttgactca
     2581 actgataaag cggacctgag gttaatctac ttggctcttg cccatatgat aaagttccgt
     2641 gggcactttc tcattgaggg tgatctaaat ccggacaact cggatgtcga caaactgttc
     2701 atccagttag tacaaaccta taatcagttg tttgaagaga accctataaa tgcaagtggc
     2761 gtggatgcga aggctattct tagcgcccgc ctctctaaat cccgacggct agaaaacctg
     2821 atcgcacaat tacccggaga gaagaaaaat gggttgttcg gtaaccttat agcgctctca
     2881 ctaggcctga caccaaattt taagtcgaac ttcgacttag ctgaagatgc caaattgcag
     2941 cttagtaagg acacgtacga tgacgatctc gacaatctac tggcacaaat tggagatcag
     3001 tatgcggact tatttttggc tgccaaaaac cttagcgatg caatcctcct atctgacata
     3061 ctgagagtta atactgagat taccaaggcg ccgttatccg cttcaatgat caaaaggtac
     3121 gatgaacatc accaagactt gacacttctc aaggccctag tccgtcagca actgcctgag
     3181 aaatataagg aaatattctt tgatcagtcg aaaaacgggt acgcaggtta tattgacggc
     3241 ggagcgagtc aagaggaatt ctacaagttt atcaaaccca tattagagaa gatggatggg
     3301 acggaagagt tgcttgtaaa actcaatcgc gaagatctac tgcgaaagca gcggactttc
     3361 gacaacggta gcattccaca tcaaatccac ttaggcgaat tgcatgctat acttagaagg
     3421 caggaggatt tttatccgtt cctcaaagac aatcgtgaaa agattgagaa aatcctaacc
     3481 tttcgcatac cttactatgt gggacccctg gcccgaggga actctcggtt cgcatggatg
     3541 acaagaaagt ccgaagaaac gattactcca tggaattttg aggaagttgt cgataaaggt
     3601 gcgtcagctc aatcgttcat cgagaggatg accaactttg acaagaattt accgaacgaa
     3661 aaagtattgc ctaagcacag tttactttac gagtatttca cagtgtacaa tgaactcacg
     3721 aaagttaagt atgtcactga gggcatgcgt aaacccgcct ttctaagcgg agaacagaag
     3781 aaagcaatag tagatctgtt attcaagacc aaccgcaaag tgacagttaa gcaattgaaa
     3841 gaggactact ttaagaaaat tgaatgcttc gattctgtcg agatctccgg ggtagaagat
     3901 cgatttaatg cgtcacttgg tacgtatcat gacctcctaa agataattaa agataaggac
     3961 ttcctggata acgaagagaa tgaagatatc ttagaagata tagtgttgac tcttaccctc
     4021 tttgaagatc gggaaatgat tgaggaaaga ctaaaaacat acgctcacct gttcgacgat
     4081 aaggttatga aacagttaaa gaggcgtcgc tatacgggct ggggacgatt gtcgcggaaa
     4141 cttatcaacg ggataagaga caagcaaagt ggtaaaacta ttctcgattt tctaaagagc
     4201 gacggcttcg ccaataggaa ctttatgcag ctgatccatg atgactcttt aaccttcaaa
     4261 gaggatatac aaaaggcaca ggtttccgga caaggggact cattgcacga acatattgcg
     4321 aatcttgctg gttcgccagc catcaaaaag ggcatactcc agacagtcaa agtagtggat
     4381 gagctagtta aggtcatggg acgtcacaaa ccggaaaaca ttgtaatcga gatggcacgc
     4441 gaaaatcaaa cgactcagaa ggggcaaaaa aacagtcgag agcggatgaa gagaatagaa
     4501 gagggtatta aagaactggg cagccagatc ttaaaggagc atcctgtgga aaatacccaa
     4561 ttgcagaacg agaaacttta cctctattac ctacaaaatg gaagggacat gtatgttgat
     4621 caggaactgg acataaaccg tttatctgat tacgacgtcg atcacattgt accccaatcc
     4681 tttttgaagg acgattcaat cgacaataaa gtgcttacac gctcggataa gaaccgaggg
     4741 aaaagtgaca atgttccaag cgaggaagtc gtaaagaaaa tgaagaacta ttggcggcag
     4801 ctcctaaatg cgaaactgat aacgcaaaga aagttcgata acttaactaa agctgagagg
     4861 ggtggcttgt ctgaacttga caaggccgga tttattaaac gtcagctcgt ggaaacccgc
     4921 caaatcacaa agcatgttgc acagatacta gattcccgaa tgaatacgaa atacgacgag
     4981 aacgataagc tgattcggga agtcaaagta atcactttaa agtcaaaatt ggtgtcggac
     5041 ttcagaaagg attttcaatt ctataaagtt agggagataa ataactacca ccatgcgcac
     5101 gacgcttatc ttaatgccgt cgtagggacc gcactcatta agaaataccc gaagctagaa
     5161 agtgagtttg tgtatggtga ttacaaagtt tatgacgtcc gtaagatgat cgcgaaaagc
     5221 gaacaggaga taggcaaggc tacagccaaa tacttctttt attctaacat tatgaatttc
     5281 tttaagacgg aaatcactct ggcaaacgga gagatacgca aacgaccttt aattgaaacc
     5341 aatggggaga caggtgaaat cgtatgggat aagggccggg acttcgcgac ggtgagaaaa
     5401 gttttgtcca tgccccaagt caacatagta aagaaaactg aggtgcagac cggagggttt
     5461 tcaaaggaat cgattcttcc aaaaaggaat agtgataagc tcatcgctcg taaaaaggac
     5521 tgggacccga aaaagtacgg tggcttcgtg agccctacag ttgcctattc tgtcctagta
     5581 gtggcaaaag ttgagaaggg aaaatccaag aaactgaagt cagtcaaaga attattgggg
     5641 ataacgatta tggagcgctc gtcttttgaa aagaacccca tcgacttcct tgaggcgaaa
     5701 ggttacaagg aagtaaaaaa ggatctcata attaaactac caaagtatag tctgtttgag
     5761 ttagaaaatg gccgaaaacg gatgttggct agcgccggag agcttcaaaa ggggaacgaa
     5821 ctcgcactac cgtctaaata cgtgaatttc ctgtatttag cgtcccatta cgagaagttg
     5881 aaaggttcac ctgaagataa cgaacagaag caactttttg ttgagcagca caaacattat
     5941 ctcgacgaaa tcatagagca aatttcggaa ttcagtaaga gagtcatcct agctgatgcc
     6001 aatctggaca aagtattaag cgcatacaac aagcacaggg ataaacccat acgtgagcag
     6061 gcggaaaata ttatccattt gtttactctt accaacctcg gcgctccagc cgcattcaag
     6121 tattttgaca caacgataga tcgcaaacag tacagatcta ccaaggaggt gctagacgcg
     6181 acactgattc accaatccat cacgggatta tatgaaactc ggatagattt gtcacagctt
     6241 gggggtgacg gatcccccaa gaagaagagg aaagtctcga gcgactacaa agaccatgac
     6301 ggtgattata aagatcatga catcgattac aaggatgacg atgacaaggg cagcggcgcc
     6361 accaacttca gcctgctgaa gcaggccggc gacgtggagg agaaccccgg ccccatggtg
     6421 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     6481 gtaaacggcc acaagttcag cgtgtctggc gagggcgagg gcgatgccac ctacggcaag
     6541 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     6601 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     6661 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     6721 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     6781 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     6841 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     6901 aaggcgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     6961 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     7021 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     7081 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagatgca
     7141 tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc
     7201 cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg
     7261 gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc
     7321 tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag
     7381 tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag
     7441 gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg
     7501 gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa
     7561 ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct
     7621 cctcttcctc atctccgggc ctttcgacct gcagcccaag cttaccatga ccgagtacaa
     7681 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     7741 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     7801 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     7861 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     7921 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     7981 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     8041 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     8101 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     8161 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     8221 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgattct agaaagtcga
     8281 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     8341 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     8401 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     8461 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     8521 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     8581 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     8641 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     8701 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     8761 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     8821 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc gtcgacttta
     8881 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     8941 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     9001 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     9061 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     9121 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtacg
     9181 tatagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     9241 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     9301 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     9361 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     9421 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     9481 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     9541 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     9601 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     9661 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     9721 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     9781 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     9841 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     9901 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     9961 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
    10021 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
    10081 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
    10141 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
    10201 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
    10261 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
    10321 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
    10381 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
    10441 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
    10501 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
    10561 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
    10621 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
    10681 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
    10741 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
    10801 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
    10861 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
    10921 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
    10981 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
    11041 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
    11101 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
    11161 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
    11221 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
    11281 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
    11341 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
    11401 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
    11461 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
    11521 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
    11581 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
    11641 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
    11701 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
    11761 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
    11821 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
    11881 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
    11941 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
    12001 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
    12061 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
    12121 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
    12181 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
    12241 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
    12301 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
    12361 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg
    12421 caagc
//
