LOCUS       sequence-392559-       15536 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 15536)
  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 15536)
  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-ABE8e-NG
FEATURES             Location/Qualifiers
     source          1..15536
                     /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             5144..9244
                     /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             9287..9307
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9308..9331
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9341..9397
                     /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             9398..9994
                     /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     9891..9911
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10010..10598
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10063..10083)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10481..10492)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10670..10903
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10929..10946)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10935..11159
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11205..11633
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11292..11311)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11502..11523
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11642..11662)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11647..11976
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11827..11962
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11889..11908
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12024..12071
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12090..12464
                     /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    12594..12715
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12631..12650)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12685..12704
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12764..12780)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12764..12780)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12777..12799)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12788..12804
                     /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(12812..12818,12819..12836,12837..12842))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12857..12878
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12995..13012)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13166..13754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13246..13265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13925..14716,14717..14785))
                     /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     14548..14567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14786..14890)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14965..14984)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        15156..15535
                     /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 cggaaagtct ctgaggtgga
     4561 gttttcccac gagtactgga tgagacatgc cctgaccctg gccaagaggg cacgggatga
     4621 gagggaggtg cctgtgggag ccgtgctggt gctgaacaat agagtgatcg gcgagggctg
     4681 gaacagagcc atcggcctgc acgacccaac agcccatgcc gaaattatgg ccctgagaca
     4741 gggcggcctg gtcatgcaga actacagact gattgacgcc accctgtacg tgacattcga
     4801 gccttgcgtg atgtgcgccg gcgccatgat ccactctagg atcggccgcg tggtgtttgg
     4861 cgtgaggaac tcaaaaagag gcgccgcagg ctccctgatg aacgtgctga actaccccgg
     4921 catgaatcac cgcgtcgaaa ttaccgaggg aatcctggca gatgaatgtg ccgccctgct
     4981 gtgcgatttc tatcggatgc ctagacaggt gttcaatgct cagaagaagg cccagagctc
     5041 catcaactcc ggaggatcta gcggaggctc ctctggctct gagacacctg gcacaagcga
     5101 gagcgcaaca cctgaaagca gcgggggcag cagcgggggg tcagacaaga agtacagcat
     5161 cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt
     5221 gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct
     5281 gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac
     5341 cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag
     5401 caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt
     5461 ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc
     5521 ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga
     5581 caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca
     5641 cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca
     5701 gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga
     5761 cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc
     5821 ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg
     5881 cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag
     5941 caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc
     6001 cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag
     6061 agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga
     6121 gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta
     6181 caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc
     6241 cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga
     6301 ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa
     6361 cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga
     6421 agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg
     6481 catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag
     6541 aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc
     6601 cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt
     6661 gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt
     6721 gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc
     6781 catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga
     6841 ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt
     6901 caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct
     6961 ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga
     7021 ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt
     7081 gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat
     7141 caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg
     7201 cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga
     7261 catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct
     7321 ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct
     7381 cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa
     7441 ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg
     7501 catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca
     7561 gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga
     7621 actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct
     7681 gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag
     7741 cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct
     7801 gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg
     7861 cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat
     7921 cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga
     7981 caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg
     8041 gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc
     8101 ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga
     8161 gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca
     8221 ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa
     8281 gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg
     8341 cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct
     8401 gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa
     8461 agagtctatc aggcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga
     8521 ccctaagaag tacggcggct tcgtcagccc caccgtggcc tattctgtgc tggtggtggc
     8581 caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac
     8641 catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta
     8701 caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga
     8761 aaacggccgg aagagaatgc tggcctctgc cagattcctg cagaagggaa acgaactggc
     8821 cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg
     8881 ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga
     8941 cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct
     9001 ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga
     9061 gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctagggcct tcaagtactt
     9121 tgacaccacc atcgaccgga aggtgtacag gagcaccaaa gaggtgctgg acgccaccct
     9181 gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg
     9241 tgactctggc ggctcaaaaa gaaccgccga cggcagcgaa ttcgagccca agaagaagag
     9301 gaaagtcgat tacaaagacg atgacgataa gggatccggc gcaacaaact tctctctgct
     9361 gaaacaagcc ggagatgtcg aagagaatcc tggaccgacc gagtacaagc ccacggtgcg
     9421 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     9481 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     9541 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     9601 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     9661 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     9721 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     9781 agtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     9841 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     9901 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     9961 cacctggtgc atgacccgca agcccggtgc ctgaacgcgt taagtcgaca atcaacctct
    10021 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
    10081 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
    10141 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
    10201 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
    10261 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
    10321 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
    10381 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
    10441 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
    10501 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
    10561 tcagacgagt cggatctccc tttgggccgc ctccccgcgt cgactttaag accaatgact
    10621 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
    10681 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
    10741 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
    10801 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
    10861 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagggcccg tttaaacccg
    10921 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
    10981 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
    11041 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
    11101 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
    11161 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
    11221 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
    11281 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
    11341 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
    11401 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
    11461 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
    11521 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
    11581 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
    11641 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
    11701 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
    11761 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
    11821 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
    11881 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
    11941 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
    12001 attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag
    12061 tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc
    12121 tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc
    12181 gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca
    12241 gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc
    12301 tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg
    12361 ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc
    12421 cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt
    12481 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
    12541 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
    12601 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
    12661 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
    12721 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
    12781 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
    12841 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
    12901 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
    12961 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
    13021 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
    13081 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
    13141 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
    13201 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
    13261 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
    13321 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
    13381 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
    13441 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
    13501 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
    13561 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
    13621 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
    13681 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
    13741 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
    13801 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
    13861 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
    13921 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
    13981 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
    14041 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
    14101 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
    14161 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
    14221 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
    14281 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
    14341 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
    14401 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
    14461 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
    14521 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
    14581 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
    14641 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
    14701 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
    14761 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
    14821 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
    14881 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
    14941 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
    15001 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
    15061 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
    15121 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
    15181 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
    15241 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
    15301 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
    15361 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
    15421 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
    15481 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgg
//
