LOCUS       sequence-273068-       13400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a tomato-targeting sgRNA and Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13400)
  AUTHORS   Hollstein PE, Eichner LJ, Brun SN, Kamireddy A, Svensson RU, Vera 
            LI, Ross DS, Rymoff TJ, Hutchins A, Galvez HM, Williams AE, 
            Shokhirev MN, Screaton RA, Berdeaux R, Shaw RJ
  TITLE     The AMPK-Related Kinases SIK1 and SIK3 Mediate Key Tumor-Suppressive
            Effects of LKB1 in NSCLC.
  JOURNAL   Cancer Discov. 2019 Nov;9(11):1606-1627. doi: 
            10.1158/2159-8290.CD-18-1261. Epub 2019 Jul 26.
  PUBMED    31350328
REFERENCE   2  (bases 1 to 13400)
  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: 
            "10.1158/2159-8290.CD-18-1261"; journalName: "Cancer Discov"; date: 
            "2019-11"; volume: "9"; issue: "11"; pages: "1606-1627"
COMMENT     Sequence Label: pLenti-Hygro-sgTom
FEATURES             Location/Qualifiers
     source          1..13400
                     /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        2259..2334
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2396..2607
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2626..2635
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2632..6735
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2632..6735
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6736..6783
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6784..6807
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6817..6873
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     misc_feature    7870..8458
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7923..7943)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8341..8352)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8530..8763
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8789..8806)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8795..9019
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9065..9493
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9152..9171)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9362..9383
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9502..9522)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9507..9836
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9687..9822
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9749..9768
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9884..9931
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9950..10324
                     /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    10454..10575
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10491..10510)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10545..10564
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10624..10640)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10624..10640)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10637..10659)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10648..10664
                     /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(10672..10678,10679..10696,10697..10702))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10717..10738
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10855..10872)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11026..11614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11106..11125)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11785..12576,12577..12645))
                     /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     12408..12427
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12646..12750)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12825..12844)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        13020..13399
                     /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 ccacgagttc gagatcgagt tttagagcta gaaatagcaa
     2281 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
     2341 gaattcgcta gctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca
     2401 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgatccggt
     2461 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
     2521 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
     2581 cgcaacgggt ttgccgccag aacacaggac cggttctaga gcgctgccac catggacaag
     2641 aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac
     2701 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
     2761 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg
     2821 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
     2881 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
     2941 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
     3001 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
     3061 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
     3121 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
     3181 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
     3241 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa
     3301 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc
     3361 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
     3421 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
     3481 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
     3541 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
     3601 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
     3661 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt
     3721 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
     3781 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg
     3841 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
     3901 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
     3961 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
     4021 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
     4081 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
     4141 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag
     4201 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
     4261 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
     4321 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
     4381 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
     4441 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
     4501 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
     4561 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
     4621 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
     4681 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
     4741 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
     4801 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
     4861 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
     4921 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
     4981 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
     5041 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
     5101 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct
     5161 cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac
     5221 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
     5281 cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc
     5341 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
     5401 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
     5461 gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
     5521 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
     5581 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
     5641 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
     5701 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
     5761 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
     5821 gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg
     5881 cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
     5941 ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag
     6001 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
     6061 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
     6121 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
     6181 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
     6241 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga
     6301 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
     6361 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag
     6421 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
     6481 gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga
     6541 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
     6601 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
     6661 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct
     6721 cagctgggag gcgacaagcg acctgccgcc acaaagaagg ctggacaggc taagaagaag
     6781 aaagattaca aagacgatga cgataaggga tccggcgcaa caaacttctc tctgctgaaa
     6841 caagccggag atgtcgaaga gaatcctgga ccgaaaaagc ctgaactcac cgcgacgtct
     6901 gtcgagaagt ttctgatcga aaagttcgac agcgtgtccg acctgatgca gctctcggag
     6961 ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta
     7021 aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc
     7081 gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc
     7141 atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct
     7201 gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg
     7261 agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc
     7321 atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc
     7381 agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa
     7441 gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc
     7501 ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc
     7561 aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag
     7621 cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt
     7681 cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag
     7741 ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc
     7801 cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tacttcgccg atagacgcgt
     7861 taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     7921 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     7981 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     8041 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     8101 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     8161 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     8221 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     8281 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     8341 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     8401 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt
     8461 cgactttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     8521 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
     8581 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     8641 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     8701 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     8761 gcagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     8821 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     8881 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     8941 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     9001 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     9061 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     9121 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     9181 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     9241 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     9301 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     9361 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     9421 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     9481 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     9541 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     9601 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     9661 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     9721 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     9781 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     9841 tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc
     9901 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt
     9961 gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac
    10021 cgaccggctc gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga
    10081 cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc
    10141 ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac
    10201 gaacttccgg gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg
    10261 ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga
    10321 ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
    10381 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
    10441 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
    10501 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
    10561 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
    10621 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
    10681 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
    10741 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
    10801 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
    10861 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
    10921 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
    10981 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
    11041 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
    11101 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
    11161 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
    11221 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
    11281 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
    11341 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
    11401 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
    11461 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
    11521 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
    11581 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
    11641 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
    11701 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
    11761 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
    11821 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
    11881 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
    11941 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
    12001 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
    12061 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
    12121 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
    12181 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
    12241 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
    12301 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
    12361 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
    12421 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
    12481 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
    12541 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
    12601 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
    12661 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
    12721 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
    12781 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
    12841 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
    12901 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
    12961 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgcgcgttg
    13021 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
    13081 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
    13141 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
    13201 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
    13261 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
    13321 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
    13381 agtcatcgct attaccatgg
//
