LOCUS       sequence-211470-       12448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for constitutive expression of Cas9-VRERRA-P2A-GFP
            in mammalian cells (codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12448)
  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 12448)
  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-P2A-GFP-PGK-Puro
FEATURES             Location/Qualifiers
     source          1..12448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..242
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             243..423
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    470..595
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1088..1321
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1506..1550
                     /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    1734..1851
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1920..2131
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2151..2160
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2160..2183
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2190..2210
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2235..5633,5634..5636,5637..5882,5883..5885,5886..6233,
                     6234..6236,6237..6239,6240..6242,6243..6335)
                     /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             2235..6335
                     /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             6336..6383
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6393..6449
                     /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(6450..6452,6453..6455,6456..7169)
                     /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(6756..6775)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7103..7124
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        7181..7680
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7216..7239)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7613..7632
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             7701..8300
                     /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(7701..7720)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8197..8217
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8316..8904
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8369..8389)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8787..8798)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8976..9209
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9287..9408
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9324..9343)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9378..9397
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9448..9583
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9603..9622)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9604..9622)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9632..9649)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9632..9648)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9641..9663)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9790..10245
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9877..9896)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10087..10108
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10271..10375
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10376..10444,10445..11236)
                     /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(10594..10613)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11407..11995
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11896..11915
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12149..12166
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12283..12304
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12319..12324,12325..12342,12343..12349)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12357..12373
                     /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     12362..12384
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12381..12397
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12381..12397
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12416..12436
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        12418..12436
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gagctgcaag cttaatgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga
       61 tgagttagca acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag
      121 taaggtggta cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac
      181 gaaccactga attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa
      241 acgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
      301 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
      361 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
      421 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg
      481 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
      541 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
      601 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
      661 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
      721 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
      781 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
      841 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga
      901 gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag
      961 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     1021 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     1081 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     1141 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     1201 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     1261 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     1321 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     1381 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     1441 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     1501 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     1561 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     1621 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     1681 caccattatc gtttcagacc cacctcccaa ccccgagggg acccagacca caattttaaa
     1741 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
     1801 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat
     1861 tacagggaca gcagagatcc actttggcgc cggctcgagt ggctccggtg cccgtcagtg
     1921 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
     1981 cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
     2041 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
     2101 ttttcgcaac gggtttgccg ccagaacaca ggtgtcgtga cgcgggatcc gccaccatgg
     2161 attacaaaga cgatgacgat aagatggccc caaagaagaa gcggaaggtc ggtatccacg
     2221 gagtcccagc agccgacaag aagtacagca tcggcctgga catcggcacc aactctgtgg
     2281 gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca
     2341 acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg
     2401 aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga
     2461 accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct
     2521 tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc
     2581 ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc
     2641 acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg
     2701 ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg
     2761 acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg
     2821 aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga
     2881 gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc
     2941 tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg
     3001 acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca
     3061 acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt
     3121 ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc
     3181 tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag
     3241 ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga
     3301 acggctacgc cggctacatt gacggcggag ccagccagga agagttctac aagttcatca
     3361 agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg
     3421 acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg
     3481 gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc
     3541 gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca
     3601 ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga
     3661 acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca
     3721 acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt
     3781 acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc
     3841 ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc
     3901 ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact
     3961 ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc
     4021 tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg
     4081 aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga
     4141 aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca
     4201 ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca
     4261 agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga
     4321 tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg
     4381 gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca
     4441 tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg
     4501 agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca
     4561 gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga
     4621 aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc
     4681 agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg
     4741 atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc
     4801 tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga
     4861 agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt
     4921 tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca
     4981 tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact
     5041 cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca
     5101 ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg
     5161 agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc
     5221 tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg
     5281 acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact
     5341 tcttctacag caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga
     5401 tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg
     5461 gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa
     5521 agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg
     5581 ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgtcagcc
     5641 ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac
     5701 tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga
     5761 atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca
     5821 agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg
     5881 ccagggaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt
     5941 acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc
     6001 tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct
     6061 ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc
     6121 accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca
     6181 atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aaggagtaca
     6241 ggagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg
     6301 agacacggat cgacctgtct cagctgggag gcgacaagcg tcctgctgct actaagaaag
     6361 ctggtcaagc taagaaaaag aaaggcagcg gcgccaccaa cttcagcctg ctgaagcagg
     6421 ccggcgacgt ggaggagaac cccggcccca tggtgagcaa gggcgaggag ctgttcaccg
     6481 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     6541 ctggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     6601 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     6661 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     6721 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     6781 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     6841 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     6901 tctatatcat ggccgacaag cagaagaacg gcatcaaggc gaacttcaag atccgccaca
     6961 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     7021 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     7081 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     7141 ctctcggcat ggacgagctg tacaagtaag ctagcctacc gggtagggga ggcgcttttc
     7201 ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt
     7261 ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt
     7321 ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg
     7381 cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag
     7441 atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag
     7501 ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc
     7561 aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc
     7621 acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg
     7681 acctgcagcc caagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     7741 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     7801 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     7861 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     7921 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     7981 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     8041 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     8101 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     8161 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     8221 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     8281 cccgcaagcc cggtgcctga ttctagaaag tcgacaatca acctctggat tacaaaattt
     8341 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     8401 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     8461 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     8521 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     8581 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     8641 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     8701 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     8761 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     8821 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     8881 tctccctttg ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt
     8941 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     9001 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     9061 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     9121 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     9181 cttttagtca gtgtggaaaa tctctagcag tacgtatagt agttcatgtc atcttattat
     9241 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     9301 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     9361 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     9421 ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     9481 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     9541 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt
     9601 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     9661 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     9721 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     9781 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     9841 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     9901 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     9961 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
    10021 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
    10081 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
    10141 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
    10201 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
    10261 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
    10321 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
    10381 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
    10441 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
    10501 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
    10561 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
    10621 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
    10681 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
    10741 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
    10801 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
    10861 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
    10921 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
    10981 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
    11041 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
    11101 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
    11161 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
    11221 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
    11281 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
    11341 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
    11401 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
    11461 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
    11521 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
    11581 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
    11641 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
    11701 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
    11761 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
    11821 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
    11881 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
    11941 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
    12001 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
    12061 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
    12121 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
    12181 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
    12241 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
    12301 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
    12361 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
    12421 taaccctcac taaagggaac aaaagctg
//
