LOCUS       sequence-211425-       11203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for constitutive expression of Cas9-VQRRA in 
            mammalian cells (codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11203)
  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 11203)
  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-VQRRA-P2A-Puro
FEATURES             Location/Qualifiers
     source          1..11203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..233
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             234..414
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    461..586
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1079..1312
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1497..1541
                     /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    1725..1842
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1911..2122
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2142..2151
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2151..2174
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2181..2201
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2226..5624,5625..5627,5628..6224,6225..6227,6228..6230,
                     6231..6233,6234..6326)
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             2226..6326
                     /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
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6327..6374
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6390..6446
                     /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             6447..7046
                     /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(6447..6466)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6943..6963
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7062..7650
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7115..7135)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7533..7544)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7722..7955
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8033..8154
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8070..8089)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8124..8143
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8194..8329
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8349..8368)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8350..8368)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8378..8395)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8378..8394)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8387..8409)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8536..8991
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8623..8642)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8833..8854
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9017..9121
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9122..9190,9191..9982)
                     /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(9340..9359)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10153..10741
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10642..10661
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10895..10912
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11029..11050
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11065..11070,11071..11088,11089..11095)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11103..11119
                     /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     11108..11130
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11127..11143
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11127..11143
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11162..11182
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11164..11182
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gcttaatgta gtcttatgca atactcttgt agtcttgcaa catggtaacg atgagttagc
       61 aacatgcctt acaaggagag aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt
      121 acgatcgtgc cttattagga aggcaacaga cgggtctgac atggattgga cgaaccactg
      181 aattgccgca ttgcagagat attgtattta agtgcctagc tcgatacata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
      481 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
      541 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
      601 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
      661 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
      721 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
      781 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
      841 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
      901 aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga
      961 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag
     1021 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag
     1081 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga
     1141 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc
     1201 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc
     1261 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt
     1321 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta
     1381 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta
     1441 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga
     1501 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa
     1561 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa
     1621 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat
     1681 cgtttcagac ccacctccca accccgaggg gacccagacc acaattttaa aagaaaaggg
     1741 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     1801 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
     1861 agcagagatc cactttggcg ccggctcgag tggctccggt gcccgtcagt gggcagagcg
     1921 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta
     1981 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
     2041 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
     2101 cgggtttgcc gccagaacac aggtgtcgtg acgcgggatc cgccaccatg gattacaaag
     2161 acgatgacga taagatggcc ccaaagaaga agcggaaggt cggtatccac ggagtcccag
     2221 cagccgacaa gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg
     2281 tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc
     2341 ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg
     2401 aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct
     2461 gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca
     2521 gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg
     2581 gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa
     2641 agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc
     2701 acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg
     2761 acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc
     2821 ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca
     2881 gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa
     2941 acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg
     3001 aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg
     3061 cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca
     3121 tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct
     3181 ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc
     3241 ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg
     3301 ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc
     3361 tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc
     3421 ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc
     3481 acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga
     3541 tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca
     3601 gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg
     3661 aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata
     3721 agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg
     3781 tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc
     3841 tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga
     3901 ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa
     3961 tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa
     4021 ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg
     4081 tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg
     4141 cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg
     4201 gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc
     4261 tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg
     4321 acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc
     4381 tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga
     4441 cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg
     4501 tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga
     4561 gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc
     4621 ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc
     4681 gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc
     4741 atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa
     4801 gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga
     4861 agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc
     4921 tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac
     4981 agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga
     5041 acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt
     5101 ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca
     5161 actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa
     5221 agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga
     5281 agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca
     5341 gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc
     5401 ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt
     5461 ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg
     5521 tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga
     5581 tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgtcagc cccaccgtgg
     5641 cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg
     5701 tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg
     5761 actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta
     5821 agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac
     5881 tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca
     5941 gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg
     6001 aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag
     6061 tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata
     6121 agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag
     6181 cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagcagtac aggagcacca
     6241 aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga
     6301 tcgacctgtc tcagctggga ggcgacaagc gtcctgctgc tactaagaaa gctggtcaag
     6361 ctaagaaaaa gaaagctagc ggcagcggcg ccaccaactt cagcctgctg aagcaggccg
     6421 gcgacgtgga ggagaacccc ggccccatga ccgagtacaa gcccacggtg cgcctcgcca
     6481 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     6541 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     6601 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     6661 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     6721 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     6781 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggagtctcgc
     6841 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     6901 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     6961 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     7021 gcatgacccg caagcccggt gcctgaacgc gttaagtcga caatcaacct ctggattaca
     7081 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     7141 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     7201 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     7261 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     7321 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     7381 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     7441 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     7501 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     7561 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     7621 gtcggatctc cctttgggcc gcctccccgc gtcgacttta agaccaatga cttacaaggc
     7681 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     7741 ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
     7801 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     7861 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     7921 cagacccttt tagtcagtgt ggaaaatctc tagcagtacg tatagtagtt catgtcatct
     7981 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt
     8041 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     8101 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     8161 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     8221 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     8281 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac
     8341 ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc
     8401 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     8461 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     8521 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     8581 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     8641 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     8701 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     8761 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     8821 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     8881 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     8941 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     9001 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     9061 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     9121 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     9181 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     9241 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     9301 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     9361 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     9421 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     9481 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     9541 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     9601 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     9661 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     9721 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     9781 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     9841 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     9901 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     9961 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
    10021 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
    10081 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
    10141 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
    10201 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
    10261 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
    10321 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
    10381 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
    10441 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
    10501 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
    10561 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
    10621 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
    10681 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
    10741 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
    10801 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
    10861 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
    10921 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
    10981 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
    11041 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
    11101 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg
    11161 cgcaattaac cctcactaaa gggaacaaaa gctggagctg caa
//
