pLentiCRISPR-RFP657

LOCUS       pLentiCRISPR-RFP657             13540 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Derived from LentiCRISPR v1 but expresses RFP657 instead of 
            puromycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    pLentiCRISPR-RFP657
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13540)
  AUTHORS   McComb S, Aguade-Gorgorio J, Harder L, Marovca B, Cario G, Eckert C,
            Schrappe M, Stanulla M, von Stackelberg A, Bourquin JP, Bornhauser 
            BC
  TITLE     Activation of concurrent apoptosis and necroptosis by SMAC mimetics 
            for the treatment of refractory and relapsed ALL.
  JOURNAL   Sci Transl Med. 2016 May 18;8(339):339ra70. doi: 
            10.1126/scitranslmed.aad2986.
  PUBMED    27194728
REFERENCE   2  (bases 1 to 13540)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..13540
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             232..412
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1077..1310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1495..1539
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        1717..1957
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     1717..1737
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1888..1907
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        3847..3922
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    3970..4087
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4156..4367
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4387..4396
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4396..4419
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4426..4446
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4471..8571
                     /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
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             8572..8619
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8629..8685
                     /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"
     misc_feature    9400..9988
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9453..9473)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9871..9882)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10057..10290
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10368..10489
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10405..10424)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10459..10478
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10529..10664
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10591..10610
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(10684..10703)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10685..10703)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10713..10730)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10713..10729)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10722..10744)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10871..11326
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10958..10977)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11168..11189
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11352..11456
                     /gene="bla"
                     /label=AmpR promoter
     CDS             11457..12317
                     /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(11675..11694)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12488..13076
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12977..12996
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13230..13247
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13364..13385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        13400..13430
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13438..13454
                     /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     13443..13465
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13462..13478
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13462..13478
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     13497..13517
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        13499..13517
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagggga cccagagagg gcctatttcc catgattcct
     1741 tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta
     1801 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
     1861 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
     1921 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggaga cggttgtaaa
     1981 tgagcacaca aaatacacat gctaaaatat tatattctat gacctttata aaatcaacca
     2041 aaatcttctt tttaataact ttagtatcaa taattagaat ttttatgttc ctttttgcaa
     2101 acttttaata aaaatgagca aaataaaaaa acgctagttt tagtaactcg cgttgttttc
     2161 ttcaccttta ataatagcta ctccaccact tgttcctaag cggtcagctc ctgcttcaat
     2221 cattttttga gcatcttcaa atgttctaac tccaccagct gctttaacta aagcattgtc
     2281 tttaacaact gacttcatta gtttaacatc ttcaaatgtt gcacctgatt ttgaaaatcc
     2341 tgttgatgtt ttaacaaatt ctaatccagc ttcaacagct atttcacaag ctttcatgat
     2401 ttcttctttt gttaataaac aattttccat aatacattta acaacatgtg atccagctgc
     2461 tttttttaca gctttcatgt cttctaaaac taattcataa tttttgtctt ttaatgcacc
     2521 aatatttaat accatatcaa tttctgttgc accatcttta attgcttcag aaacttcgaa
     2581 tgcttttgta gctgttgtgc atgcacctag aggaaaacct acaacatttg ttattcctac
     2641 atttgtgcct tttaataatt ctttacaata gcttgttcaa tatgaattaa cacaaactgt
     2701 tgcaaaatca aattcaattg cttcatcaca taattgttta atttcagctt tcgtagcatc
     2761 ttgttttaat aatgtgtgat ctatatattt gtttagtttc attttttctc ctatatattc
     2821 atttttaatt ttaattcttt aataatttcg tctactttaa ctttagcgtt ttgaacagat
     2881 tcaccaacac ctataaaata aatttttagt ttaggttcag ttccacttgg gcgaacagca
     2941 aatcatgact tatcttctaa ataaaatttt agtaagtctt gtcctggcat attatacatt
     3001 ccatcgatgt agtcttcaac attaacaact ttaagtccag caatttgagt taagggtgtt
     3061 gctctcaatg atttcattaa tggttcaatt tttaatttct tttcttctgg tttaaaattc
     3121 aagtttaaag tgaaagtgta atatgcaccc atttctttaa ataaatcttc taaatagtct
     3181 actaatgttt tattttgttt tttataaaat caagcagcct ctgctattaa tatagaagct
     3241 tgtattccat ctttatctct agctgagtca tcaattacat atccataact ttcttcataa
     3301 gcaaaaacaa aatttaatcc gttatcttct tctttagcaa tttctctacc cattcattta
     3361 aatccagtta aagtttttac aatattaact ccatattttt catgagcgat tctatcaccc
     3421 aaatcacttg ttacaaaact tgaatataga gccggatttt ttggaatgct atttaagcgt
     3481 tttagatttg ataattttca atcaattaaa attggtcctg tttgatttcc atctaatctt
     3541 acaaaatgac catcatgttt tattgccatt ccaaatctgt cagcatctgg gtcattcata
     3601 ataataatat ctgcatcatg tttaatacca tattcaagcg gtatttttca tgcaggatca
     3661 aattctggat ttggatttac aacattttta aatgtttcat cttcaaatgc atgctcttca
     3721 acctcaataa cgttatatcc tgattcacgt aatatttttg gggtaaattt agttcctgtt
     3781 ccattaactg cgctaaaaat aatttttaaa tcttttttag cttcttgctc ttttttgtac
     3841 gtctctgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
     3901 aaaagtggca ccgagtcggt gcttttttga attctagatc ttgagacaaa tggcagtatt
     3961 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     4021 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     4081 aaattttcgg gtttattaca gggacagcag agatccactt tggcgccggc tcgagtggct
     4141 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     4201 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     4261 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     4321 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggtg tcgtgacgcg
     4381 ggatccgcca ccatggatta caaagacgat gacgataaga tggccccaaa gaagaagcgg
     4441 aaggtcggta tccacggagt cccagcagcc gacaagaagt acagcatcgg cctggacatc
     4501 ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa
     4561 ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg
     4621 ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga
     4681 tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc
     4741 aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag
     4801 aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag
     4861 taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg
     4921 cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag
     4981 ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc
     5041 tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc
     5101 ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc
     5161 gagaagaaga atggcctgtt cggcaacctg attgccctga gcctgggcct gacccccaac
     5221 ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac
     5281 gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg
     5341 gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag
     5401 atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac
     5461 ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc
     5521 ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag
     5581 ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg
     5641 aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc
     5701 caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca
     5761 ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac
     5821 gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa
     5881 accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc
     5941 atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac
     6001 agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc
     6061 gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg
     6121 ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa
     6181 atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg
     6241 ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa
     6301 aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg
     6361 atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg
     6421 aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg
     6481 gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga
     6541 aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc
     6601 caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc
     6661 gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg
     6721 ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag
     6781 aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg
     6841 ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg
     6901 tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac
     6961 cggctgtccg actacgatgt ggaccatatc gtgcctcaga gctttctgaa ggacgactcc
     7021 atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc
     7081 tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg
     7141 attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg
     7201 gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg
     7261 gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg
     7321 gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag
     7381 ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc
     7441 gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc
     7501 gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag
     7561 gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc
     7621 ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga aaccggggag
     7681 atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa
     7741 gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatcctg
     7801 cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac
     7861 ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag
     7921 ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga
     7981 agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa
     8041 aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag
     8101 agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct gccctccaaa
     8161 tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat
     8221 aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag
     8281 cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg
     8341 tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac
     8401 ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc
     8461 gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc
     8521 atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga caagcgtcct
     8581 gctgctacta agaaagctgg tcaagctaag aaaaagaaag ctagcggcgc aacaaacttc
     8641 tctctgctga aacaagccgg agatgtcgaa gagaatcctg gaccgagcga gctgattaag
     8701 gagaacatgc acatgaagct gtacatggag ggcaccgtga acaaccacca cttcaagtgc
     8761 acatccgagg gcgaaggcaa gccctacgag ggcacccaga cccagagaat caaggtggtc
     8821 gagggcggcc ctctcccctt cgccttcgac atcctggcta ccagcttcat gtacggcagt
     8881 cacaccttca tcaaccacac ccagggcatc cccgacttct ggaagcagtc cttccctgag
     8941 ggcttcacat gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag
     9001 gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc
     9061 ccatccaacg gccctgtgat gcagaagaaa acactcggct gggaggccca caccgagatg
     9121 ctgtaccccg ctgacggcgg cctggaaggc agaaccgcgc tggccctgaa gctcgtgggc
     9181 gggggccacc tgatctgcaa cttcaagacc acatacagat ccaagaaacc cgctaagaac
     9241 ctcaagatgc ccggcgtcta ctatgtggac tacagactgg aaagaatcaa ggaggccgac
     9301 aaagagacct acgtcgagca gcacgaggtg gctgtggcca gatactgcga cctccctagc
     9361 aaactggggc acaagcttaa ttaaacgcgt taagtcgaca atcaacctct ggattacaaa
     9421 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     9481 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     9541 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     9601 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     9661 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     9721 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     9781 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     9841 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     9901 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     9961 cggatctccc tttgggccgc ctccccgcga ctttaagacc aatgacttac aaggcagctg
    10021 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
    10081 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
    10141 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
    10201 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
    10261 ccttttagtc agtgtggaaa atctctagca gtacgtatag tagttcatgt catcttatta
    10321 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
    10381 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
    10441 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
    10501 tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
    10561 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
    10621 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat
    10681 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
    10741 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
    10801 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
    10861 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
    10921 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
    10981 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
    11041 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
    11101 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
    11161 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
    11221 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
    11281 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
    11341 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
    11401 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
    11461 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
    11521 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
    11581 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
    11641 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
    11701 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
    11761 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
    11821 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
    11881 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
    11941 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
    12001 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
    12061 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
    12121 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
    12181 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
    12241 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
    12301 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
    12361 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
    12421 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
    12481 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
    12541 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
    12601 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
    12661 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
    12721 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
    12781 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
    12841 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
    12901 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
    12961 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
    13021 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
    13081 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
    13141 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
    13201 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
    13261 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
    13321 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
    13381 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
    13441 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa
    13501 ttaaccctca ctaaagggaa caaaagctgg agctgcaagc
//