LOCUS       sequence-396570-       16220 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus vector for base editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16220)
  AUTHORS   Li H, Ma T, Remsberg JR, Won SJ, DeMeester KE, Njomen E, Ogasawara 
            D, Zhao KT, Huang TP, Lu B, Simon GM, Melillo B, Schreiber SL, 
            Lykke-Andersen J, Liu DR, Cravatt BF
  TITLE     Assigning functionality to cysteines by base editing of cancer 
            dependency genes.
  JOURNAL   Nat Chem Biol. 2023 Nov;19(11):1320-1330. doi: 
            10.1038/s41589-023-01428-w. Epub 2023 Oct 2.
  PUBMED    37783940
REFERENCE   2  (bases 1 to 16220)
  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"; doi: 
            "10.1038/s41589-023-01428-w"; journalName: "Nat Chem Biol"; date: 
            "2023-11"; volume: "19"; issue: "11"; pages: "1320-1330"
COMMENT     Sequence Label: All-in-one lenti-evoCDA-NG
FEATURES             Location/Qualifiers
     source          1..16220
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..199
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     154..174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             217..397
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    444..569
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1062..1295
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1480..1524
                     /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    1822..1939
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1990..2238
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1990..2010
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2161..2180
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4120..4195
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4257..4468
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             4529..4549
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5270..9370
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SIRPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARFLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPRAFKYFDTTIDRKVYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             9401..9649
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             9680..9928
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             9971..9991
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9992..10015
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             10025..10081
                     /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             10082..10678
                     /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     10575..10595
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10694..11282
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10747..10767)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11165..11176)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11354..11587
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(11613..11630)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11619..11843
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11889..12317
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11976..11995)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12186..12207
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12326..12346)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12331..12660
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12511..12646
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12573..12592
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12708..12755
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12774..13148
                     /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    13278..13399
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13315..13334)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13369..13388
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13448..13464)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13448..13464)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13461..13483)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13472..13488
                     /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(13496..13502,13503..13520,13521..13526))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13541..13562
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13679..13696)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13850..14438)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13930..13949)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14609..15400,15401..15469))
                     /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     15232..15251
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15470..15574)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(15649..15668)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        15840..16219
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
      241 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
      301 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
      361 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
      421 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
      481 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
      541 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
      601 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
      661 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
      721 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
      781 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
      841 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
      901 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
      961 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1021 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1081 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1141 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1201 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1261 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1321 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1381 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1441 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1501 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1561 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1621 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1681 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1741 agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca
     1801 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     1861 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     1921 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggttaat
     1981 taaggtaccg agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct
     2041 gttagagaga taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg
     2101 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg
     2161 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg
     2221 tggaaaggac gaaacaccgg agacggttgt aaatgagcac acaaaataca catgctaaaa
     2281 tattatattc tatgaccttt ataaaatcaa ccaaaatctt ctttttaata actttagtat
     2341 caataattag aatttttatg ttcctttttg caaactttta ataaaaatga gcaaaataaa
     2401 aaaacgctag ttttagtaac tcgcgttgtt ttcttcacct ttaataatag ctactccacc
     2461 acttgttcct aagcggtcag ctcctgcttc aatcattttt tgagcatctt caaatgttct
     2521 aactccacca gctgctttaa ctaaagcatt gtctttaaca actgacttca ttagtttaac
     2581 atcttcaaat gttgcacctg attttgaaaa tcctgttgat gttttaacaa attctaatcc
     2641 agcttcaaca gctatttcac aagctttcat gatttcttct tttgttaata aacaattttc
     2701 cataatacat ttaacaacat gtgatccagc tgcttttttt acagctttca tgtcttctaa
     2761 aactaattca taatttttgt cttttaatgc accaatattt aataccatat caatttctgt
     2821 tgcaccatct ttaattgctt cagaaacttc gaatgctttt gtagctgttg tgcatgcacc
     2881 tagaggaaaa cctacaacat ttgttattcc tacatttgtg ccttttaata attctttaca
     2941 atagcttgtt caatatgaat taacacaaac tgttgcaaaa tcaaattcaa ttgcttcatc
     3001 acataattgt ttaatttcag ctttcgtagc atcttgtttt aataatgtgt gatctatata
     3061 tttgtttagt ttcatttttt ctcctatata ttcattttta attttaattc tttaataatt
     3121 tcgtctactt taactttagc gttttgaaca gattcaccaa cacctataaa ataaattttt
     3181 agtttaggtt cagttccact tgggcgaaca gcaaatcatg acttatcttc taaataaaat
     3241 tttagtaagt cttgtcctgg catattatac attccatcga tgtagtcttc aacattaaca
     3301 actttaagtc cagcaatttg agttaagggt gttgctctca atgatttcat taatggttca
     3361 atttttaatt tcttttcttc tggtttaaaa ttcaagttta aagtgaaagt gtaatatgca
     3421 cccatttctt taaataaatc ttctaaatag tctactaatg ttttattttg ttttttataa
     3481 aatcaagcag cctctgctat taatatagaa gcttgtattc catctttatc tctagctgag
     3541 tcatcaatta catatccata actttcttca taagcaaaaa caaaatttaa tccgttatct
     3601 tcttctttag caatttctct acccattcat ttaaatccag ttaaagtttt tacaatatta
     3661 actccatatt tttcatgagc gattctatca cccaaatcac ttgttacaaa acttgaatat
     3721 agagccggat tttttggaat gctatttaag cgttttagat ttgataattt tcaatcaatt
     3781 aaaattggtc ctgtttgatt tccatctaat cttacaaaat gaccatcatg ttttattgcc
     3841 attccaaatc tgtcagcatc tgggtcattc ataataataa tatctgcatc atgtttaata
     3901 ccatattcaa gcggtatttt tcatgcagga tcaaattctg gatttggatt tacaacattt
     3961 ttaaatgttt catcttcaaa tgcatgctct tcaacctcaa taacgttata tcctgattca
     4021 cgtaatattt ttggggtaaa tttagttcct gttccattaa ctgcgctaaa aataattttt
     4081 aaatcttttt tagcttcttg ctcttttttg tacgtctctg ttttagagct agaaatagca
     4141 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
     4201 tgaattcgct agctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc
     4261 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgatccgg
     4321 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     4381 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     4441 tcgcaacggg tttgccgcca gaacacagga ccggttctag agcgctgcca ccatgaaacg
     4501 gacagccgac ggaagcgagt tcgagtcacc aaagaagaag cggaaagtca gtaccgacgc
     4561 cgagtacgtg cggatccacg agaagctgga tatctataca ttcaagaagc agtttagcaa
     4621 caataagaag tccgtgtctc acagatgcta cgtgctgttc gagctgaagc ggagaggaga
     4681 gaggcgcgcc tgtttttggg gctatgccgt gaacaagcca cagtctggaa ccgagagggg
     4741 aatccacgca gagatcttca gcatcaggaa ggtggaggag tacctgcgcg acaaccccgg
     4801 ccagtttaca atcaattggt atagctcctg gagcccttgc gccgattgtg ccgagaagat
     4861 cctggagtgg tacaaccagg agctgagggg caatggccac accctgaaga tctgggtgtg
     4921 caagctgtac tatgagaaga acgccaggaa tcagatcggc ctgtggaacc tgcgcgacaa
     4981 tggcgtgggc ctgaacgtga tggtgtccga gcactatcag tgctgtcgca agatctttat
     5041 ccagtctagc cacaatcagc tgaacgagaa tcggtggctg gagaagacac tgaagagagc
     5101 cgagaagcgg agaagcgagc tgtccatcat gtttcaggtg aagatcctgc acaccacaaa
     5161 gtctcccgcc gtgtctggcg gatctagcgg aggatcctct ggcagcgaga caccaggaac
     5221 aagcgagtca gcaacaccag agagcagtgg cggcagcagc ggcggcagcg acaagaagta
     5281 cagcatcggc ctggccatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta
     5341 caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa
     5401 gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa
     5461 gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat
     5521 cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt
     5581 cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga
     5641 ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag
     5701 caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg
     5761 gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt
     5821 catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg
     5881 cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct
     5941 gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag
     6001 cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca
     6061 gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca
     6121 gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat
     6181 cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata
     6241 cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga
     6301 gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg
     6361 cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg
     6421 caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt
     6481 cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg
     6541 gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac
     6601 cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat
     6661 gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg
     6721 cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga
     6781 gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac
     6841 caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa
     6901 aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa
     6961 agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga
     7021 tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga
     7081 cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact
     7141 gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga
     7201 caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa
     7261 gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc
     7321 cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa
     7381 agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc
     7441 caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga
     7501 cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag
     7561 agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga
     7621 agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca
     7681 gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga
     7741 ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag
     7801 ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg
     7861 caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca
     7921 gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag
     7981 aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg
     8041 gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga
     8101 gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga
     8161 tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca
     8221 cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga
     8281 aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag
     8341 cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt
     8401 tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac
     8461 aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa
     8521 agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt
     8581 cagcaaagag tctatccggc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga
     8641 ctgggaccct aagaagtacg gcggcttcgt gagccccacc gtggcctatt ctgtgctggt
     8701 ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg
     8761 gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa
     8821 gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga
     8881 gctggaaaac ggccggaaga gaatgctggc ctctgcccgg ttcctgcaga agggaaacga
     8941 actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct
     9001 gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta
     9061 cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc
     9121 taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca
     9181 ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctc gggccttcaa
     9241 gtactttgac accaccatcg accggaaggt gtaccggagc accaaagagg tgctggacgc
     9301 caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct
     9361 gggaggtgac agcggcggga gcggcgggag cggggggagc actaatctga gcgacatcat
     9421 tgagaaggag actgggaaac agctggtcat tcaggagtcc atcctgatgc tgcctgagga
     9481 ggtggaggaa gtgatcggca acaagccaga gtctgacatc ctggtgcaca ccgcctacga
     9541 cgagtccaca gatgagaatg tgatgctgct gacctctgac gcccccgagt ataagccttg
     9601 ggccctggtc atccaggatt ctaacggcga gaataagatc aagatgctga gcggaggatc
     9661 cggaggatct ggaggcagca ccaacctgtc tgacatcatc gagaaggaga caggcaagca
     9721 gctggtcatc caggagagca tcctgatgct gcccgaagaa gtcgaagaag tgatcggaaa
     9781 caagcctgag agcgatatcc tggtccatac cgcctacgac gagagtaccg acgaaaatgt
     9841 gatgctgctg acatccgacg ccccagagta taagccctgg gctctggtca tccaggattc
     9901 caacggagag aacaaaatca aaatgctgtc tggcggctca aaaagaaccg ccgacggcag
     9961 cgaattcgag cccaagaaga agaggaaagt cgattacaaa gacgatgacg ataagggatc
    10021 cggcgcaaca aacttctctc tgctgaaaca agccggagat gtcgaagaga atcctggacc
    10081 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
    10141 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
    10201 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
    10261 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
    10321 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
    10381 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
    10441 cgcgtggttc ctggccaccg tcggagtctc gcccgaccac cagggcaagg gtctgggcag
    10501 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
    10561 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
    10621 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaac
    10681 gcgttaagtc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
    10741 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
    10801 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
    10861 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
    10921 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
    10981 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
    11041 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
    11101 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
    11161 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
    11221 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
    11281 gcgtcgactt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
    11341 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
    11401 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
    11461 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
    11521 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
    11581 tctagcaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
    11641 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
    11701 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
    11761 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
    11821 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
    11881 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
    11941 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
    12001 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
    12061 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
    12121 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
    12181 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
    12241 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
    12301 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
    12361 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
    12421 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
    12481 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
    12541 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
    12601 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
    12661 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat
    12721 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca
    12781 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
    12841 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
    12901 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
    12961 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
    13021 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
    13081 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
    13141 aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
    13201 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
    13261 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
    13321 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
    13381 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
    13441 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
    13501 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
    13561 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
    13621 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
    13681 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
    13741 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
    13801 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
    13861 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
    13921 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
    13981 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
    14041 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
    14101 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
    14161 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
    14221 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
    14281 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
    14341 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
    14401 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
    14461 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
    14521 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
    14581 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
    14641 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
    14701 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
    14761 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
    14821 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
    14881 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
    14941 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
    15001 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
    15061 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
    15121 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
    15181 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
    15241 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
    15301 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
    15361 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
    15421 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
    15481 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
    15541 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
    15601 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
    15661 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
    15721 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
    15781 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
    15841 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
    15901 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
    15961 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
    16021 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
    16081 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
    16141 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
    16201 agtcatcgct attaccatgg
//
