LOCUS       sequence-211412-       11188 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for constitutive expression of Cas9-VQR (not codon
            optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11188)
  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 11188)
  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"; journalName: "Nat 
            Biotechnol. 2018 Jul 3. pii: nbt.4194. doi: 10.1038/nbt.4194."
COMMENT     Sequence Label: pLenti-VQR-P2A-Puro
FEATURES             Location/Qualifiers
     source          1..11188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             222..402
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    449..574
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1067..1300
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1485..1529
                     /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    1713..1830
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1899..2110
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2130..2139
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2136..5537,5538..5540,5541..6137,6138..6140,6141..6143,
                     6144..6146,6147..6239)
                     /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             6246..6266
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6273..6338
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6363..6419
                     /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             6420..7019
                     /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(6420..6439)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6916..6936
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7035..7623
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7088..7108)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7506..7517)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7695..7928
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8006..8127
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8043..8062)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8097..8116
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8167..8302
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8322..8341)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8323..8341)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8351..8368)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8351..8367)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8360..8382)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8509..8964
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8596..8615)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8806..8827
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8990..9094
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9095..9163,9164..9955)
                     /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(9313..9332)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10126..10714
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10615..10634
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10868..10885
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11002..11023
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11038..11043,11044..11061,11062..11068)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11076..11092
                     /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     11081..11103
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11100..11116
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11100..11116
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11135..11155
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11137..11155
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        join(11183..11188,1..221)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa catgccttac
       61 aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac gatcgtgcct
      121 tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa ttgccgcatt
      181 gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc tggttagacc
      241 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
      301 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
      361 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
      421 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
      481 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
      541 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
      601 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
      661 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
      721 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
      781 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
      841 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
      901 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
      961 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     1021 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     1081 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     1141 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     1201 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     1261 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     1321 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     1381 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     1441 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     1501 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     1561 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     1621 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     1681 acctcccaac cccgagggga cccagaccac aattttaaaa gaaaaggggg gattgggggg
     1741 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     1801 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
     1861 ctttggcgcc ggctcgagtg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac
     1921 agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg
     1981 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg
     2041 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc
     2101 cagaacacag gtgtcgtgac gcgggatccg ccaccatgga taaaaagtat tctattggtt
     2161 tagacatcgg cactaattcc gttggatggg ctgtcataac cgatgaatac aaagtacctt
     2221 caaagaaatt taaggtgttg gggaacacag accgtcattc gattaaaaag aatcttatcg
     2281 gtgccctcct attcgatagt ggcgaaacgg cagaggcgac tcgcctgaaa cgaaccgctc
     2341 ggagaaggta tacacgtcgc aagaaccgaa tatgttactt acaagaaatt tttagcaatg
     2401 agatggccaa agttgacgat tctttctttc accgtttgga agagtccttc cttgtcgaag
     2461 aggacaagaa acatgaacgg caccccatct ttggaaacat agtagatgag gtggcatatc
     2521 atgaaaagta cccaacgatt tatcacctca gaaaaaagct agttgactca actgataaag
     2581 cggacctgag gttaatctac ttggctcttg cccatatgat aaagttccgt gggcactttc
     2641 tcattgaggg tgatctaaat ccggacaact cggatgtcga caaactgttc atccagttag
     2701 tacaaaccta taatcagttg tttgaagaga accctataaa tgcaagtggc gtggatgcga
     2761 aggctattct tagcgcccgc ctctctaaat cccgacggct agaaaacctg atcgcacaat
     2821 tacccggaga gaagaaaaat gggttgttcg gtaaccttat agcgctctca ctaggcctga
     2881 caccaaattt taagtcgaac ttcgacttag ctgaagatgc caaattgcag cttagtaagg
     2941 acacgtacga tgacgatctc gacaatctac tggcacaaat tggagatcag tatgcggact
     3001 tatttttggc tgccaaaaac cttagcgatg caatcctcct atctgacata ctgagagtta
     3061 atactgagat taccaaggcg ccgttatccg cttcaatgat caaaaggtac gatgaacatc
     3121 accaagactt gacacttctc aaggccctag tccgtcagca actgcctgag aaatataagg
     3181 aaatattctt tgatcagtcg aaaaacgggt acgcaggtta tattgacggc ggagcgagtc
     3241 aagaggaatt ctacaagttt atcaaaccca tattagagaa gatggatggg acggaagagt
     3301 tgcttgtaaa actcaatcgc gaagatctac tgcgaaagca gcggactttc gacaacggta
     3361 gcattccaca tcaaatccac ttaggcgaat tgcatgctat acttagaagg caggaggatt
     3421 tttatccgtt cctcaaagac aatcgtgaaa agattgagaa aatcctaacc tttcgcatac
     3481 cttactatgt gggacccctg gcccgaggga actctcggtt cgcatggatg acaagaaagt
     3541 ccgaagaaac gattactcca tggaattttg aggaagttgt cgataaaggt gcgtcagctc
     3601 aatcgttcat cgagaggatg accaactttg acaagaattt accgaacgaa aaagtattgc
     3661 ctaagcacag tttactttac gagtatttca cagtgtacaa tgaactcacg aaagttaagt
     3721 atgtcactga gggcatgcgt aaacccgcct ttctaagcgg agaacagaag aaagcaatag
     3781 tagatctgtt attcaagacc aaccgcaaag tgacagttaa gcaattgaaa gaggactact
     3841 ttaagaaaat tgaatgcttc gattctgtcg agatctccgg ggtagaagat cgatttaatg
     3901 cgtcacttgg tacgtatcat gacctcctaa agataattaa agataaggac ttcctggata
     3961 acgaagagaa tgaagatatc ttagaagata tagtgttgac tcttaccctc tttgaagatc
     4021 gggaaatgat tgaggaaaga ctaaaaacat acgctcacct gttcgacgat aaggttatga
     4081 aacagttaaa gaggcgtcgc tatacgggct ggggacgatt gtcgcggaaa cttatcaacg
     4141 ggataagaga caagcaaagt ggtaaaacta ttctcgattt tctaaagagc gacggcttcg
     4201 ccaataggaa ctttatgcag ctgatccatg atgactcttt aaccttcaaa gaggatatac
     4261 aaaaggcaca ggtttccgga caaggggact cattgcacga acatattgcg aatcttgctg
     4321 gttcgccagc catcaaaaag ggcatactcc agacagtcaa agtagtggat gagctagtta
     4381 aggtcatggg acgtcacaaa ccggaaaaca ttgtaatcga gatggcacgc gaaaatcaaa
     4441 cgactcagaa ggggcaaaaa aacagtcgag agcggatgaa gagaatagaa gagggtatta
     4501 aagaactggg cagccagatc ttaaaggagc atcctgtgga aaatacccaa ttgcagaacg
     4561 agaaacttta cctctattac ctacaaaatg gaagggacat gtatgttgat caggaactgg
     4621 acataaaccg tttatctgat tacgacgtcg atcacattgt accccaatcc tttttgaagg
     4681 acgattcaat cgacaataaa gtgcttacac gctcggataa gaaccgaggg aaaagtgaca
     4741 atgttccaag cgaggaagtc gtaaagaaaa tgaagaacta ttggcggcag ctcctaaatg
     4801 cgaaactgat aacgcaaaga aagttcgata acttaactaa agctgagagg ggtggcttgt
     4861 ctgaacttga caaggccgga tttattaaac gtcagctcgt ggaaacccgc caaatcacaa
     4921 agcatgttgc acagatacta gattcccgaa tgaatacgaa atacgacgag aacgataagc
     4981 tgattcggga agtcaaagta atcactttaa agtcaaaatt ggtgtcggac ttcagaaagg
     5041 attttcaatt ctataaagtt agggagataa ataactacca ccatgcgcac gacgcttatc
     5101 ttaatgccgt cgtagggacc gcactcatta agaaataccc gaagctagaa agtgagtttg
     5161 tgtatggtga ttacaaagtt tatgacgtcc gtaagatgat cgcgaaaagc gaacaggaga
     5221 taggcaaggc tacagccaaa tacttctttt attctaacat tatgaatttc tttaagacgg
     5281 aaatcactct ggcaaacgga gagatacgca aacgaccttt aattgaaacc aatggggaga
     5341 caggtgaaat cgtatgggat aagggccggg acttcgcgac ggtgagaaaa gttttgtcca
     5401 tgccccaagt caacatagta aagaaaactg aggtgcagac cggagggttt tcaaaggaat
     5461 cgattcttcc aaaaaggaat agtgataagc tcatcgctcg taaaaaggac tgggacccga
     5521 aaaagtacgg tggcttcgtg agccctacag ttgcctattc tgtcctagta gtggcaaaag
     5581 ttgagaaggg aaaatccaag aaactgaagt cagtcaaaga attattgggg ataacgatta
     5641 tggagcgctc gtcttttgaa aagaacccca tcgacttcct tgaggcgaaa ggttacaagg
     5701 aagtaaaaaa ggatctcata attaaactac caaagtatag tctgtttgag ttagaaaatg
     5761 gccgaaaacg gatgttggct agcgccggag agcttcaaaa ggggaacgaa ctcgcactac
     5821 cgtctaaata cgtgaatttc ctgtatttag cgtcccatta cgagaagttg aaaggttcac
     5881 ctgaagataa cgaacagaag caactttttg ttgagcagca caaacattat ctcgacgaaa
     5941 tcatagagca aatttcggaa ttcagtaaga gagtcatcct agctgatgcc aatctggaca
     6001 aagtattaag cgcatacaac aagcacaggg ataaacccat acgtgagcag gcggaaaata
     6061 ttatccattt gtttactctt accaacctcg gcgctccagc cgcattcaag tattttgaca
     6121 caacgataga tcgcaaacag tacagatcta ccaaggaggt gctagacgcg acactgattc
     6181 accaatccat cacgggatta tatgaaactc ggatagattt gtcacagctt gggggtgacg
     6241 gatcccccaa gaagaagagg aaagtctcga gcgactacaa agaccatgac ggtgattata
     6301 aagatcatga catcgattac aaggatgacg atgacaaggc tgcaggagct agcggcagcg
     6361 gcgccaccaa cttcagcctg ctgaagcagg ccggcgacgt ggaggagaac cccggcccca
     6421 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     6481 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     6541 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     6601 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     6661 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     6721 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     6781 ccgcgtggtt cctggccacc gtcggagtct cgcccgacca ccagggcaag ggtctgggca
     6841 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     6901 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     6961 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa
     7021 cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     7081 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     7141 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     7201 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     7261 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     7321 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     7381 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     7441 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     7501 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     7561 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     7621 cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     7681 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc
     7741 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     7801 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     7861 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     7921 ctctagcagt acgtatagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     7981 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     8041 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     8101 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     8161 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     8221 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     8281 tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt
     8341 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     8401 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     8461 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     8521 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     8581 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     8641 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     8701 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     8761 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     8821 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     8881 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     8941 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     9001 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     9061 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     9121 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     9181 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     9241 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     9301 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     9361 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     9421 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     9481 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     9541 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     9601 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     9661 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     9721 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     9781 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     9841 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     9901 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     9961 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
    10021 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
    10081 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
    10141 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
    10201 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
    10261 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
    10321 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
    10381 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
    10441 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
    10501 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
    10561 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
    10621 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
    10681 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
    10741 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
    10801 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
    10861 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
    10921 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
    10981 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
    11041 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
    11101 aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca
    11161 aaagctggag ctgcaagctt aatgtagt
//
