LOCUS       sequence-392558-       16001 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 16001)
  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 16001)
  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-BE4-NG
FEATURES             Location/Qualifiers
     source          1..16001
                     /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             4550..5233
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG
                     RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS
                     RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE
                     AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL
                     WATGLK"
     CDS             5330..9430
                     /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             9461..9709
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             9752..9772
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9773..9796
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9806..9862
                     /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             9863..10459
                     /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     10356..10376
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10475..11063
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10528..10548)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10946..10957)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11135..11368
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(11394..11411)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11400..11624
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11670..12098
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11757..11776)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11967..11988
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12107..12127)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12112..12441
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12292..12427
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12354..12373
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12489..12536
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12555..12929
                     /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    13059..13180
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13096..13115)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13150..13169
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13229..13245)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13229..13245)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13242..13264)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13253..13269
                     /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(13277..13283,13284..13301,13302..13307))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13322..13343
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13460..13477)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13631..14219)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13711..13730)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14390..15181,15182..15250))
                     /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     15013..15032
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15251..15355)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(15430..15449)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        15621..16000
                     /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 gttcagagac
     4561 tgggcctgtc gccgtcgatc caaccctgcg ccgccggatt gaacctcacg agtttgaagt
     4621 gttctttgac ccccgggagc tgagaaagga gacatgcctg ctgtacgaga tcaactgggg
     4681 aggcaggcac tccatctgga ggcacacctc tcagaacaca aataagcacg tggaggtgaa
     4741 cttcatcgag aagtttacca cagagcggta cttctgcccc aataccagat gtagcatcac
     4801 atggtttctg agctggtccc cttgcggaga gtgtagcagg gccatcaccg agttcctgtc
     4861 cagatatcca cacgtgacac tgtttatcta catcgccagg ctgtatcacc acgcagaccc
     4921 aaggaatagg cagggcctgc gcgatctgat cagctccggc gtgaccatcc agatcatgac
     4981 agagcaggag tccggctact gctggcggaa cttcgtgaat tattctccta gcaacgaggc
     5041 ccactggcct aggtacccac acctgtgggt gcgcctgtac gtgctggagc tgtattgcat
     5101 catcctgggc ctgccccctt gtctgaatat cctgcggaga aagcagcccc agctgacctt
     5161 ctttacaatc gccctgcagt cttgtcacta tcagaggctg ccaccccaca tcctgtgggc
     5221 cacaggcctg aagtctggcg gatctagcgg aggatcctct ggcagcgaga caccaggaac
     5281 aagcgagtca gcaacaccag agagcagtgg cggcagcagc ggcggcagcg acaagaagta
     5341 cagcatcggc ctggccatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta
     5401 caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa
     5461 gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa
     5521 gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat
     5581 cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt
     5641 cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga
     5701 ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag
     5761 caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg
     5821 gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt
     5881 catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg
     5941 cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct
     6001 gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag
     6061 cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca
     6121 gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca
     6181 gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat
     6241 cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata
     6301 cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga
     6361 gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg
     6421 cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg
     6481 caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt
     6541 cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg
     6601 gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac
     6661 cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat
     6721 gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg
     6781 cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga
     6841 gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac
     6901 caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa
     6961 aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa
     7021 agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga
     7081 tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga
     7141 cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact
     7201 gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga
     7261 caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa
     7321 gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc
     7381 cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa
     7441 agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc
     7501 caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga
     7561 cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag
     7621 agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga
     7681 agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca
     7741 gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga
     7801 ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag
     7861 ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg
     7921 caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca
     7981 gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag
     8041 aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg
     8101 gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga
     8161 gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga
     8221 tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca
     8281 cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga
     8341 aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag
     8401 cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt
     8461 tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac
     8521 aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa
     8581 agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt
     8641 cagcaaagag tctatccggc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga
     8701 ctgggaccct aagaagtacg gcggcttcgt gagccccacc gtggcctatt ctgtgctggt
     8761 ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg
     8821 gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa
     8881 gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga
     8941 gctggaaaac ggccggaaga gaatgctggc ctctgcccgg ttcctgcaga agggaaacga
     9001 actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct
     9061 gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta
     9121 cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc
     9181 taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca
     9241 ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctc gggccttcaa
     9301 gtactttgac accaccatcg accggaaggt gtaccggagc accaaagagg tgctggacgc
     9361 caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct
     9421 gggaggtgac agcggcggga gcggcgggag cggggggagc accaacctgt ctgacatcat
     9481 cgagaaggag acaggcaagc agctggtcat ccaggagagc atcctgatgc tgcccgaaga
     9541 agtcgaagaa gtgatcggaa acaagcctga gagcgatatc ctggtccata ccgcctacga
     9601 cgagagtacc gacgaaaatg tgatgctgct gacatccgac gccccagagt ataagccctg
     9661 ggctctggtc atccaggatt ccaacggaga gaacaaaatc aaaatgctgt ctggcggctc
     9721 aaaaagaacc gccgacggca gcgaattcga gcccaagaag aagaggaaag tcgattacaa
     9781 agacgatgac gataagggat ccggcgcaac aaacttctct ctgctgaaac aagccggaga
     9841 tgtcgaagag aatcctggac cgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     9901 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     9961 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
    10021 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
    10081 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
    10141 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
    10201 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggagtct cgcccgacca
    10261 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
    10321 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
    10381 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
    10441 ccgcaagccc ggtgcctgaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg
    10501 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
    10561 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
    10621 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
    10681 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
    10741 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
    10801 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
    10861 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
    10921 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
    10981 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
    11041 ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta
    11101 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
    11161 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
    11221 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
    11281 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
    11341 ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat cagcctcgac
    11401 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
    11461 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
    11521 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
    11581 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
    11641 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
    11701 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
    11761 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
    11821 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
    11881 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
    11941 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
    12001 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
    12061 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
    12121 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
    12181 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
    12241 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
    12301 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
    12361 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
    12421 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
    12481 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
    12541 tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
    12601 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
    12661 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
    12721 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
    12781 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
    12841 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
    12901 tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg
    12961 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
    13021 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
    13081 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
    13141 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
    13201 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
    13261 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
    13321 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
    13381 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
    13441 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
    13501 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
    13561 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
    13621 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
    13681 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
    13741 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
    13801 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
    13861 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
    13921 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
    13981 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
    14041 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
    14101 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
    14161 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
    14221 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
    14281 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
    14341 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
    14401 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
    14461 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
    14521 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
    14581 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
    14641 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
    14701 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
    14761 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
    14821 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
    14881 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
    14941 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
    15001 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
    15061 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
    15121 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
    15181 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
    15241 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
    15301 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
    15361 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg
    15421 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
    15481 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
    15541 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
    15601 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
    15661 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
    15721 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
    15781 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
    15841 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
    15901 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
    15961 ttggcagtac atctacgtat tagtcatcgc tattaccatg g
//
