LOCUS       sequence-105912-       13792 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lenti vector for expression of inducible split Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13792)
  AUTHORS   Zetsche B, Volz SE, Zhang F
  TITLE     A split-Cas9 architecture for inducible genome editing and 
            transcription modulation.
  JOURNAL   Nat Biotechnol. 2015 Feb 2. doi: 10.1038/nbt.3149.
  PUBMED    25643054
REFERENCE   2  (bases 1 to 13792)
  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. 2015 Feb 2. doi: 10.1038/nbt.3149."
COMMENT     Sequence Label: pLSC-5
FEATURES             Location/Qualifiers
     source          1..13792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2608..2856
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2608..2628
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2779..2798
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2873..2948
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3010..3221
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3229..3238
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3241..3261
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3286..3606
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             3652..6036
                     /codon_start=1
                     /product="C-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(C)
                     /translation="CFDSVEISGVEDRFNASLGAYHDLLKIIKDKDFLDNEENEDILED
                     IVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGK
                     TILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKG
                     ILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQI
                     LKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDN
                     KVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDK
                     AGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQF
                     YKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGK
                     ATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMP
                     QVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKV
                     EKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELEN
                     GRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLD
                     EIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKY
                     FDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD"
     CDS             6037..6084
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6094..6150
                     /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             6175..7890
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     CDS             7936..8205
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQ"
     CDS             8215..8271
                     /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             8272..8868
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     8765..8785
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8884..9472
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8937..8957)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9355..9366)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9544..9777
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9803..9820)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9809..10033
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10079..10507
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10166..10185)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10376..10397
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10516..10536)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10521..10850
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10701..10836
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10763..10782
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10898..10945
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10964..11338
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    11468..11589
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11505..11524)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11559..11578
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11638..11654)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11638..11654)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11651..11673)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11662..11678
                     /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)."
     promoter        complement(join(11686..11692,11693..11710,11711..11716))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11731..11752
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11869..11886)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12040..12628)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12120..12139)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12799..13590,13591..13659))
                     /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     13422..13441
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13660..13764)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta aggtaccgag ggcctatttc ccatgattcc ttcatatttg
     2641 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag
     2701 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
     2761 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
     2821 ttggctttat atatcttgtg gaaaggacga aacaccggag acggacgtct ctgttttaga
     2881 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
     2941 gtcggtgctt ttttgaattc gctagctagg tcttgaaagg agtgggaatt ggctccggtg
     3001 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
     3061 gcaattgatc cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
     3121 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
     3181 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggaccggtgc caccatggcc
     3241 ccaaagaaga agcggaaggt cggtatccac ggagtcccag cagccggcgt gcaggtggag
     3301 accatcagcc ccggcgacgg cagaaccttc cccaagagag gccagacctg cgtggtgcac
     3361 tacaccggca tgctggagga cggcaagaag ttcgacagca gcagagacag aaacaagccc
     3421 ttcaagttca tgctgggcaa gcaggaggtg atcagaggct gggaggaggg cgtggcccag
     3481 atgagcgtgg gccagagagc caagctgacc atcagccccg actacgccta cggcgccacc
     3541 ggccaccccg gcatcatccc tcctcacgcc accctggtgt tcgacgtgga gctgctgaag
     3601 ctggagggtg gcggtggctc gggtggcggt ggctcgggtg gcggtggctc gtgcttcgac
     3661 tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcgc ataccacgat
     3721 ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg
     3781 gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg
     3841 aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac
     3901 accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc
     3961 aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg
     4021 atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag
     4081 ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc
     4141 atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc
     4201 gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac
     4261 agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg
     4321 aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg
     4381 cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac
     4441 gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg
     4501 ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg
     4561 aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag
     4621 ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc
     4681 atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac
     4741 tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc
     4801 accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc
     4861 gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc
     4921 ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac
     4981 gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac
     5041 ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag
     5101 atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag
     5161 ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa
     5221 aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc
     5281 gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc
     5341 cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa
     5401 ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag
     5461 aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc
     5521 aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct
     5581 gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg
     5641 tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag
     5701 ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc
     5761 tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag
     5821 caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc
     5881 aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac
     5941 accagcacca aagaggtgct ggatgccacc ctgatccacc agagcatcac cggcctgtac
     6001 gagacacgga tcgacctgtc tcagctggga ggcgacaaaa ggccggcggc cacgaaaaag
     6061 gccggccagg caaaaaagaa aaagggatcc ggcgcaacaa acttctctct gctgaaacaa
     6121 gccggagatg tcgaagagaa tcctggaccg ctagatttag ctagcttaat cctcgacaag
     6181 aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac
     6241 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
     6301 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg
     6361 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
     6421 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
     6481 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
     6541 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
     6601 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
     6661 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
     6721 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
     6781 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa
     6841 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc
     6901 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
     6961 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
     7021 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
     7081 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
     7141 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
     7201 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt
     7261 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
     7321 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctacg gaagcagcgg
     7381 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
     7441 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
     7501 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
     7561 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
     7621 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
     7681 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag
     7741 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
     7801 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
     7861 ctgaaagagg actacttcaa gaaaatcgag ggtggcggtg gctcgggtgg cggtggctcg
     7921 ggtggcggtg gctcggagat gtggcacgag ggcctggagg aggccagcag actgtacttc
     7981 ggcgagagaa acgtgaaggg catgttcgag gtgctggagc ccctgcacgc catgatggag
     8041 agaggccccc agaccctgaa ggagaccagc ttcaaccagg cctacggcag agacctgatg
     8101 gaggcccagg agtggtgcag aaagtacatg aagagcggca acgtgaagga cctgacccag
     8161 gcctgggacc tgtactacca cgtgttcaga agaatcagca agcagggatc cggcgcaaca
     8221 aacttctctc tgctgaaaca agccggagat gtcgaagaga atcctggacc gaccgagtac
     8281 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     8341 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     8401 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     8461 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     8521 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     8581 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     8641 ctggccaccg tcggagtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     8701 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     8761 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     8821 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaac gcgttaagtc
     8881 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     8941 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     9001 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     9061 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     9121 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     9181 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     9241 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     9301 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     9361 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     9421 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcgtcgactt
     9481 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     9541 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
     9601 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     9661 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     9721 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaggg
     9781 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     9841 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     9901 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     9961 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
    10021 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
    10081 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
    10141 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
    10201 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
    10261 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
    10321 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
    10381 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
    10441 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
    10501 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
    10561 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
    10621 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
    10681 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
    10741 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
    10801 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
    10861 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
    10921 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
    10981 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
    11041 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
    11101 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
    11161 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
    11221 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
    11281 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
    11341 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
    11401 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
    11461 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
    11521 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
    11581 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
    11641 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
    11701 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
    11761 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
    11821 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
    11881 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
    11941 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
    12001 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
    12061 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
    12121 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
    12181 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
    12241 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
    12301 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
    12361 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
    12421 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
    12481 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
    12541 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
    12601 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
    12661 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
    12721 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
    12781 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
    12841 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
    12901 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
    12961 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
    13021 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
    13081 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
    13141 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
    13201 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
    13261 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
    13321 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
    13381 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
    13441 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
    13501 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
    13561 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
    13621 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
    13681 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
    13741 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ac
//
