LOCUS       sequence-211464-       11665 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for constitutive expression of Cas9-VRERRA in 
            mammalian cells (codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11665)
  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 11665)
  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-VRERRA-PGK-Puro
FEATURES             Location/Qualifiers
     source          1..11665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             224..404
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    451..576
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1069..1302
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1487..1531
                     /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    1715..1832
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1901..2112
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2132..2141
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2141..2164
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2171..2191
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2216..5614,5615..5617,5618..5863,5864..5866,5867..6214,
                     6215..6217,6218..6220,6221..6223,6224..6316)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/G1218R/R1335E/T1337R mutations"
                     /label=Cas9 VRER
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKEYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             2216..6316
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKEYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6317..6364
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     promoter        6379..6878
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6414..6437)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6811..6830
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6899..7498
                     /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(6899..6918)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7395..7415
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7514..8102
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7567..7587)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7985..7996)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8174..8407
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8485..8606
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8522..8541)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8576..8595
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8646..8781
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8801..8820)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8802..8820)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8830..8847)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8830..8846)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8839..8861)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8988..9443
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9075..9094)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9285..9306
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9469..9573
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9574..9642,9643..10434)
                     /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(9792..9811)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10605..11193
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11094..11113
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11347..11364
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11481..11502
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11517..11522,11523..11540,11541..11547)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11555..11571
                     /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     11560..11582
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11579..11595
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11579..11595
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11614..11634
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11616..11634
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        join(11662..11665,1..223)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gtcttatgca atactcttgt agtcttgcaa catggtaacg atgagttagc aacatgcctt
       61 acaaggagag aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc
      121 cttattagga aggcaacaga cgggtctgac atggattgga cgaaccactg aattgccgca
      181 ttgcagagat attgtattta agtgcctagc tcgatacata aacgggtctc tctggttaga
      241 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
      301 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
      361 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
      421 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
      481 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
      541 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
      601 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
      661 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
      721 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
      781 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
      841 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc
      901 accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg
      961 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac
     1021 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt
     1081 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
     1141 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
     1201 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
     1261 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
     1321 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
     1381 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     1441 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     1501 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
     1561 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
     1621 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
     1681 ccacctccca accccgaggg gacccagacc acaattttaa aagaaaaggg gggattgggg
     1741 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
     1801 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc
     1861 cactttggcg ccggctcgag tggctccggt gcccgtcagt gggcagagcg cacatcgccc
     1921 acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg
     1981 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
     2041 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
     2101 gccagaacac aggtgtcgtg acgcgggatc cgccaccatg gattacaaag acgatgacga
     2161 taagatggcc ccaaagaaga agcggaaggt cggtatccac ggagtcccag cagccgacaa
     2221 gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga
     2281 cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat
     2341 caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg
     2401 gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca
     2461 agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga
     2521 gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt
     2581 ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt
     2641 ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa
     2701 gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa
     2761 gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc
     2821 cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga
     2881 aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc
     2941 cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa
     3001 actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg
     3061 cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag
     3121 cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa
     3181 gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct
     3241 gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat
     3301 tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat
     3361 ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg
     3421 gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct
     3481 gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat
     3541 cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc
     3601 ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga
     3661 caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc
     3721 caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga
     3781 gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga
     3841 gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca
     3901 gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt
     3961 ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga
     4021 caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct
     4081 gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt
     4141 cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag
     4201 ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct
     4261 gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac
     4321 ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca
     4381 cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt
     4441 ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat
     4501 ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg
     4561 gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa
     4621 cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta
     4681 cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc
     4741 tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa
     4801 ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg
     4861 gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc
     4921 cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga
     4981 aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta
     5041 cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt
     5101 gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca
     5161 cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa
     5221 gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc
     5281 caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat
     5341 gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat
     5401 cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt
     5461 gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg
     5521 cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa
     5581 gaaggactgg gaccctaaga agtacggcgg cttcgtcagc cccaccgtgg cctattctgt
     5641 gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct
     5701 gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga
     5761 agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct
     5821 gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccagggaac tgcagaaggg
     5881 aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga
     5941 gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa
     6001 gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc
     6061 cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag
     6121 agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc
     6181 cttcaagtac tttgacacca ccatcgaccg gaaggagtac aggagcacca aagaggtgct
     6241 ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc
     6301 tcagctggga ggcgacaagc gtcctgctgc tactaagaaa gctggtcaag ctaagaaaaa
     6361 gaaataagct agcctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg
     6421 ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt
     6481 ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt
     6541 ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt
     6601 gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat
     6661 ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg
     6721 ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg
     6781 cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg
     6841 ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttaccat
     6901 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     6961 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     7021 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     7081 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     7141 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     7201 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     7261 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     7321 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     7381 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     7441 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgatt
     7501 ctagaaagtc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     7561 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     7621 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     7681 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     7741 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     7801 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     7861 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     7921 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     7981 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     8041 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     8101 gcgtcgactt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     8161 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
     8221 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     8281 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     8341 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     8401 tctagcagta cgtatagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     8461 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
     8521 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     8581 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
     8641 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     8701 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     8761 gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta
     8821 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     8881 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     8941 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     9001 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     9061 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     9121 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     9181 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     9241 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     9301 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     9361 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     9421 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     9481 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     9541 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     9601 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     9661 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     9721 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     9781 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     9841 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     9901 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     9961 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
    10021 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
    10081 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
    10141 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
    10201 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
    10261 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
    10321 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
    10381 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
    10441 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
    10501 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
    10561 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
    10621 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
    10681 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
    10741 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
    10801 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
    10861 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
    10921 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
    10981 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
    11041 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
    11101 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
    11161 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
    11221 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
    11281 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
    11341 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
    11401 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
    11461 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
    11521 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
    11581 ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa
    11641 aagctggagc tgcaagctta atgta
//
