LOCUS       sequence-338714-       16976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  doxycycline inducible BE4max YE1 NG expression for human CLYBL locus
            knock-in.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16976)
  AUTHORS   Coelho MA, Cooper S, Strauss ME, Karakoc E, Bhosle S, Goncalves E, 
            Picco G, Burgold T, Cattaneo CM, Veninga V, Consonni S, Dincer C, 
            Vieira SF, Gibson F, Barthorpe S, Hardy C, Rein J, Thomas M, Marioni
            J, Voest EE, Bassett A, Garnett MJ
  TITLE     Base editing screens map mutations affecting interferon-gamma 
            signaling in cancer.
  JOURNAL   Cancer Cell. 2023 Feb 13;41(2):288-303.e6. doi: 
            10.1016/j.ccell.2022.12.009. Epub 2023 Jan 19.
  PUBMED    36669486
REFERENCE   2  (bases 1 to 16976)
  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: 
            "10.1016/j.ccell.2022.12.009"; journalName: "Cancer Cell"; date: 
            "2023-02-13- 13"; volume: "41"; issue: "2"; pages: "288-303.e6"
COMMENT     Sequence Label: iBE4max YE1 NG CLYBL
FEATURES             Location/Qualifiers
     source          1..16976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     266..285
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2052..2429
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    2059..2077
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2095..2113
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2131..2149
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2167..2185
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2203..2221
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2239..2257
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2275..2293
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2338..2362
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2481..2501
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3282..7382
                     /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             7413..7661
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7692..7940
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7983..8003
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    8037..8158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8074..8093)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8128..8147
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8207..8223)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8207..8223)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8220..8242)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8231..8247
                     /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(8255..8261,8262..8279,8280..8285))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8300..8321
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8438..8455)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     8747..8766
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    8765..8820
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     8866..8885
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(8976..9722)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(9754..9773)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          9781..10798
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(9835..9857)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(10800..11077)
                     /label=chicken beta-actin promoter
     enhancer        11079..11458
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    11464..11688
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     11677..11694
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    11710..11743
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      12521..12530
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(12579..12605)
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             complement(12621..12647)
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             complement(12654..12707)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     LTR             13176..13444
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(13457..13668)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     polyA_signal    13704..13928
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(13933..14733)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="GPRIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(14044..14063)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     14654..14673
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(14728..14781)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    14805..14830
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    14831..14864
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             join(15722..15790,15791..16582)
                     /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     complement(15940..15959)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(16753..16976,1..365)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
       61 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      121 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      181 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      241 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      301 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      361 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc gaattcgccc ttagagcaga
      421 gccaggaacc cctgtaggga aggggcagga gagccagggg catgagatgg tggacgagga
      481 agggggacag ggaagcctga gcgcctctcc tgggcttgcc aaggactcaa acccagaagc
      541 ccagagcagg gccttaggga agcgggacgg ctctgttgga gagtccagta ttgaattagt
      601 gatagataca cagaaaaaaa gcaaattgaa taggtgaatg gataaacaaa ctgtggaaca
      661 tccagacaat gaaatattca ccaataaaaa gaaatgagct atcaagccac attggctatg
      721 cccttcacat ttaaggagga accttaaatg catattgcca aattttaaaa gctagtctga
      781 aaatgctaca gactgtatga ttccaattat atgacatttt agaaaaggca aaactataga
      841 gatcagtggt tgccagcggc tcatggaggc aggatgaatt ggtggaacac agggatgctt
      901 tcagggtaat gaaaatatct tgtgtgatcc tgtaataatg gatacgtggc ataatgcatt
      961 tgttaaaacc tgtaaactag ccaggtatag tggctttttt gcacctatag tctcagctac
     1021 ttggtaggct gagacaggaa gatcacttga gcccaggact tcaagaccag cctgggtaat
     1081 atagtaagac cccatctcta ttttttaaag aaattaagaa agaaaagaaa cccatcaact
     1141 gtataacaca aagaatcgta atgtaaacta cagtcctgag taatgaaaca tttttaaaga
     1201 aataaataaa ttcaaggcaa ttattaactt caggaaaaac agtaaagtaa acataagtat
     1261 acaaatcaaa catggctcag ttgtgaaaat atttacaagg tcactttaag taacattgaa
     1321 tgttgatata atcccaaaat atatttatgt tggaaggatg aggagattta attaagggag
     1381 gtggtccctg cagttacgcc aatgataacc cccgccagaa aaatcttagt agccttccct
     1441 ttttgttttc cgtgccccaa ctcggcggat tgactcggcc ccttccggaa acacccgaat
     1501 caacttctag tcaaattatt gttcacgccg caatgaccca cccctggccc gcgtctgtgg
     1561 aactgacccc tggtgtacag gagagttcgc tgctgaaagt ggtcccaaag gggtactagt
     1621 ttttaagctc ccaactcccc ctcccccagc gtctggagga ttccacaccc tcgcaccgca
     1681 ggggcgagga agtgggcgga gtccggtttt ggcgccagcc gctgaggctg ccaagcagaa
     1741 aagccaccgc tgaggagact ccggtcactg tcctcgcccc gcctccccct tccctcccct
     1801 tggggaccac cgggcgccac gccgcgaacg gtaagtgccg cggtcgtcgg cgcctccgcc
     1861 ctccccctag ggccccaatt cccagcgggc gcggcgcgcg gcccctcccc ccgccgggcg
     1921 cgcgcccgct gccccgccct tcgtggccgc ccggcgtggg cggtgccacc cctccccccg
     1981 gcggccccgc gcgcagctcc cggctccctc ccccttcgga tgtggcttga gctgtaggcg
     2041 cggagggtac cagtttactc cctatcagtg atagagaacg tatgaagagt ttactcccta
     2101 tcagtgatag agaacgtatg cagactttac tccctatcag tgatagagaa cgtataagga
     2161 gtttactccc tatcagtgat agagaacgta tgaccagttt actccctatc agtgatagag
     2221 aacgtatcta cagtttactc cctatcagtg atagagaacg tatatccagt ttactcccta
     2281 tcagtgatag agaacgtata agctttaggc gtgtacggtg ggcgcctata aaagcagagc
     2341 tcgtttagtg aaccgtcaga tcgcctggag caattccaca acacttttgt cttataccaa
     2401 ctttccgtac cacttcctac cctcgtaaag gatccgccgc caccatgaaa cggacagccg
     2461 acggaagcga gttcgagtca ccaaagaaga agcggaaagt ctcctcagag actgggcctg
     2521 tcgccgtcga tccaaccctg cgccgccgga ttgaacctca cgagtttgaa gtgttctttg
     2581 acccccggga gctgagaaag gagacatgcc tgctgtacga gatcaactgg ggaggcaggc
     2641 actccatctg gaggcacacc tctcagaaca caaataagca cgtggaggtg aacttcatcg
     2701 agaagtttac cacagagcgg tacttctgcc ccaataccag atgtagcatc acatggtttc
     2761 tgagctattc cccttgcgga gagtgtagca gggccatcac cgagttcctg tccagatatc
     2821 cacacgtgac actgtttatc tacatcgcca ggctgtatca ccacgcagac ccagagaata
     2881 ggcagggcct gcgcgatctg atcagctccg gcgtgaccat ccagatcatg acagagcagg
     2941 agtccggcta ctgctggcgg aacttcgtga attattctcc tagcaacgag gcccactggc
     3001 ctaggtaccc acacctgtgg gtgcgcctgt acgtgctgga gctgtattgc atcatcctgg
     3061 gcctgccccc ttgtctgaat atcctgcgga gaaagcagcc ccagctgacc ttctttacaa
     3121 tcgccctgca gtcttgtcac tatcagaggc tgccacccca catcctgtgg gccacaggcc
     3181 tgaagtctgg aggatctagc ggaggatcct ctggcagcga gacaccagga acaagcgagt
     3241 cagcaacacc agagagcagt ggcggcagca gcggcggcag cgacaagaag tacagcatcg
     3301 gcctggccat cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc
     3361 ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga
     3421 tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg
     3481 ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca
     3541 acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg
     3601 aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct
     3661 accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca
     3721 aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact
     3781 tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc
     3841 tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg
     3901 ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc
     3961 agctgcccgg cgagaagaag aatggcctgt tcggaaacct gattgccctg agcctgggcc
     4021 tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca
     4081 aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg
     4141 acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag
     4201 tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc
     4261 accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca
     4321 aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca
     4381 gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg
     4441 aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg
     4501 gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag
     4561 atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca
     4621 tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa
     4681 agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgcttccg
     4741 cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc
     4801 tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga
     4861 aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca
     4921 tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact
     4981 acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca
     5041 acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg
     5101 acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg
     5161 acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga
     5221 tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca
     5281 acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct
     5341 tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca
     5401 tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg
     5461 ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg
     5521 tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc
     5581 agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca
     5641 tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga
     5701 acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac
     5761 tggacatcaa ccggctgtcc gactacgatg tggaccatat cgtgcctcag agctttctga
     5821 aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg ggcaagagcg
     5881 acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga
     5941 acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc
     6001 tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca
     6061 caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca
     6121 agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga
     6181 aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct
     6241 acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt
     6301 tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg
     6361 aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga
     6421 ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg
     6481 aaaccgggga gatcgtgtgg gataagggcc gggattttgc caccgtgcgg aaagtgctga
     6541 gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag
     6601 agtctatcag gcccaagagg aacagcgata agctgatcgc cagaaagaag gactgggacc
     6661 ctaagaagta cggcggcttc gtcagcccca ccgtggccta ttctgtgctg gtggtggcca
     6721 aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca
     6781 tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca
     6841 aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa
     6901 acggccggaa gagaatgctg gcctctgcca gattcctgca gaagggaaac gaactggccc
     6961 tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct
     7021 cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaagcac tacctggacg
     7081 agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg
     7141 acaaagtgct gtccgcctac aacaagcacc gggataagcc catcagagag caggccgaga
     7201 atatcatcca cctgtttacc ctgaccaatc tgggagcccc tagggccttc aagtactttg
     7261 acaccaccat cgaccggaag gtgtacagga gcaccaaaga ggtgctggac gccaccctga
     7321 tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggtg
     7381 acagcggcgg gagcggcggg agcgggggga gcactaatct gagcgacatc attgagaagg
     7441 agactgggaa acagctggtc attcaggagt ccatcctgat gctgcctgag gaggtggagg
     7501 aagtgatcgg caacaagcca gagtctgaca tcctggtgca caccgcctac gacgagtcca
     7561 cagatgagaa tgtgatgctg ctgacctctg acgcccccga gtataagcct tgggccctgg
     7621 tcatccagga ttctaacggc gagaataaga tcaagatgct gagcggagga tccggaggat
     7681 ctggaggcag caccaacctg tctgacatca tcgagaagga gacaggcaag cagctggtca
     7741 tccaggagag catcctgatg ctgcccgaag aagtcgaaga agtgatcgga aacaagcctg
     7801 agagcgatat cctggtccat accgcctacg acgagagtac cgacgaaaat gtgatgctgc
     7861 tgacatccga cgccccagag tataagccct gggctctggt catccaggat tccaacggag
     7921 agaacaaaat caaaatgctg tctggcggct caaaaagaac cgccgacggc agcgaattcg
     7981 agcccaagaa gaagaggaaa gtctaagttt aaactcgcga atcgatcgcc caccccaact
     8041 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     8101 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     8161 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     8221 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     8281 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     8341 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     8401 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     8461 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggacggccgc acatgtaagg
     8521 aaaattttag ggatgttaaa gaaaaaaata acacaaaaca aaatataaaa aaaatctaac
     8581 ctcaagtcaa ggcttttcta tggaataagg aatggacagc agggggctgt ttcatatact
     8641 gatgacctct ttatagccaa cctttgttca tggcagccag catatgggca tatgttgcca
     8701 aactctaaac caaatactca ttctgatgtt ttaaatgatt tgccctccca tatgtccttc
     8761 cgagtgagag acacaaaaaa ttccaacaca ctattgcaat gaaaataaat ttcctttatt
     8821 agccagaagt cagatgctca aggggcttca tgatgtcccc ataatttttg gcagagggaa
     8881 aaagatctca gtggtatttg tgagccaggg cattggccac accagccacc accttctgat
     8941 aggcagcctg cacctgagga gtgaattcac gcgttttagc ctggcagcat atcgagatcg
     9001 aagtcatcga gagcgtcagc agggagcata tccagatcaa aatcatccag ggcatcggcg
     9061 ggcagcatgt ccaggtcgaa gtcgtccagg gcgtcggtgg gtccgccgct ctcgcacttc
     9121 agctgcttct cgaggccgca gatgatcagt tccaggccga acaggaaggc gggctcggcg
     9181 ccctgtctgt cgaacagctc gatggcctgc ttcagcaggg ggggcatgct gtcggtggtg
     9241 ggtgtctctc tttcctcttt ggcgacctgg tgttcctgtt cttccagcac gcagcccagg
     9301 gtgaagtggc ccacggcgct cagggcgtac agggcgtttt ccaggctgaa gccctgctgg
     9361 cacaggaagg ccagctggtt ttccagtgtc tcgtactgct tctcggtggg tctggtgccc
     9421 aggtgcactt tggcgccgtc ccggtggctc agcagggcgc atctgtagct cttggcgttg
     9481 ttccgcagga agtcctgcca gctctcgcct tccagagggc agctgtgggt gtggtgccgg
     9541 tccagcatct cgatgggcag ggcgtccagc agggcccgct tgttcttcac gtgccagtac
     9601 agggtgggct gttccacgcc cagtttctgg gccagcttcc gggtggtcag gccctcgatg
     9661 cccacgccgt tcagcagttc cagggcgctg ttgatcactt tgctcttgtc cagccggctc
     9721 atgtcgacat tctttgccaa aatgatgaga cagcacaata accagcacgt tgcccaggag
     9781 ctgtaggaaa aagaagaagg catgaacatg gttagcagag gctctagagc cgccggtcac
     9841 acgccagaag ccgaaccccg ccctgccccg tcccccccga aggcagccgt ccccccgcgg
     9901 acagccccga ggctggagag ggagaagggg acggcggcgc ggcgacgcac gaaggccctc
     9961 cccgcccatt tccttcctgc cggcgccgca ccgcttcgcc ccgcgcccgc tagagggggt
    10021 gcggcggcgc ctcccagatt tcggctccgc acagatttgg gacaaaggaa gtccctgcgc
    10081 cctctcgcac gattaccata aaaggcaatg gctgcggctc gccgcgcctc gacagccgcc
    10141 ggcgctccgg gggccgccgc gcccctcccc cgagccctcc ccggcccgag gcggccccgc
    10201 cccgcccggc acccccacct gccgccaccc cccgcccggc acggcgagcc ccgcgccacg
    10261 ccccgtacgg agccccgcac ccgaagccgg gccgtgctca gcaactcggg gaggggggtg
    10321 cagggggggt tgcagcccga ccgacgcgcc cacaccccct gctcaccccc ccacgcacac
    10381 accccgcacg cagcctttgt tcccctcgca gccccccccg caccgcgggg caccgccccc
    10441 ggccgcgctc ccctcgcgca cactgcggag cgcacaaagc cccgcgccgc gcccgcagcg
    10501 ctcacagccg ccgggcagcg cggagccgca cgcggcgctc cccacgcaca cacacacgca
    10561 cgcacccccc gagccgctcc ccccgcacaa agggccctcc cggagcccct caaggctttc
    10621 acgcagccac agaaaagaaa caagccgtca ttaaaccaag cgctaattac agcccggagg
    10681 agaagggccg tcccgcccgc tcacctgtgg gagtaacgcg gtcagtcaga gccggggcgg
    10741 gcggcgcgag gcggcggcgg agcggggcac ggggcgaagg cagcgtcgca gcgactcccc
    10801 gcccgccgcg cgcttcgctt tttatagggc cgccgccgcc gccgcctcgc cataaaagga
    10861 aactttcgga gcgcgccgct ctgattggct gccgccgcac ctctccgcct cgccccgccc
    10921 cgcccctcgc cccgccccgc cccgcctggc gcgcgccccc cccccccccc cgcccccatc
    10981 gctgcacaaa ataattaaaa aataaataaa tacaaaattg ggggtgggga ggggggggag
    11041 atggggagag tgaagcagaa cgtggggctc acctcgacca tggtaatagc gatgactaat
    11101 acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag
    11161 gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga
    11221 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
    11281 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
    11341 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
    11401 ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcga
    11461 gacccataga gcccaccgca tccccagcat gcctgctatt gtcttcccaa tcctccccct
    11521 tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga caatgcgatg
    11581 caatttcctc attttattag gaaaggacag tgggagtggc accttccagg gtcaaggaag
    11641 gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagtc gaggctgatc
    11701 agcgggttta taacttcgta taatgtatgc tatacgaagt tatggtacct tactcgtcca
    11761 tgccgccggt ggagtggcgg ccctcggcgc gttcgtactg ttccacgatg gtgttacttg
    11821 tacagctcgt ccattccgcc tgtgctgtgt ctgccttcgg ctctctcgta ctgttccacg
    11881 atggtgtagt cctcgttgtg ggacacgatg tccagcttga tatccacgat gtaggcgcca
    11941 ggcagctgca cgggtttctt ggccttgtag gtggttttca cttcggcggc atagtggccg
    12001 ccatccttca gcttcagccg cttcttgatc tcggacttca gggcgccatc ctcagggtac
    12061 attctttcct cgctggcttc ccagcccatg gtctttttct gcatcacggg gccatcagag
    12121 ggaaagttgg tgccccgcag cttcactttg tagatgaaca cgccgtcctg cagagagcta
    12181 tcctggttca cgtggatgat gccgccgtcc tcgaagttca tcactctctc ccatctgaag
    12241 ccctcgggga agctcagctt gaagtagtcg ggaatgtcgg cggggtgctt gatgtacacc
    12301 ttgctgccgt acataaactg agggctcaga atgtcccagg caaaaggcag agggccgcct
    12361 ttggtcactt tcagcttggc ggtctgaaaa gcctcgtagg gtctgccctc gccttcgcct
    12421 tcaatctcga actcgtggcc gttcacgctg ccttccatgt gcaccttgaa ccgcatgaat
    12481 tctttgatga tggccatgtt gttctcttcg cccttggaca ccatggtggc gaccggtagc
    12541 gtggcgaccg gggtggcggt ggctccccct gaccctccat ccaacttaac gcgctttgca
    12601 gcaggacctc cagaaccgcc gtcgagtttg acccgtttag cggcagggac cggtgggcca
    12661 ggattctcct cgacgtcacc gcatgttagc agacttcctc tgccctctcc cgatcccgat
    12721 ccatttcggg tatatttgag tggaatgagt tcttcaatcg tagttttgac taacttgcca
    12781 ttcatttcta ttaacacaaa acaatctggt gcatagtctg aaatcaactc cctacacata
    12841 ccacaaggac ttaccactcg aatacttcta tctacttcgt cagaataagg gtgtctaaca
    12901 gctacaatcg tgtcaaaatc cttttgtcca ttcgaaactg cactaccaat cgcaatggct
    12961 tctgcacaaa cagttactcg tcctatatac gcttcaatat gtactgccga aatgatttct
    13021 cctgttttcg tacgaattgc cgctcccaca tgatgtttat tatcctcata aagcattgta
    13081 atcttctctg tcgctacttc tactaattct agatcctgct gggagatgtt gaaggtcttc
    13141 atggtggcgc tagcgtaggc gccggtcaca gcttggatct gtaacggcgc agaacagaaa
    13201 acgaaacaaa gacgtagagt tgagcaagca gggtcaggca aagcgtggag agccggctga
    13261 gtctaggtag gctccaaggg agcgccggac aaaggcccgg tctcgacctg agctttaaac
    13321 ttacctagac ggcggacgca gttcaggagg caccacaggc gggaggcggc agaacgcgac
    13381 tcaaccggcg tggatggcgg cctcaggtag ggcggcgggc gcgtgaagga gagatgcgag
    13441 cccctcgaag cttcagctgt gttctggcgg caaacccgtt gcgaaaaaga acgttcacgg
    13501 cgactactgc acttatatac ggttctcccc caccctcggg aaaaaggcgg agccagtaca
    13561 cgacatcact ttcccagttt accccgcgcc accttctcta ggcaccggtt caattgccga
    13621 cccctccccc caacttctcg gggactgtgg gcgatgtgcg ctctgcccac tgacgggcac
    13681 cggagcgatc gcagatcatg catccataga gcccaccgca tccccagcat gcctgctatt
    13741 gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca
    13801 cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc
    13861 accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga
    13921 aggcacaggt ttttagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc
    13981 gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc
    14041 agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc
    14101 acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc
    14161 gccatgggtc acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag
    14221 ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc
    14281 ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt
    14341 agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc
    14401 aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc
    14461 ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag
    14521 ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt
    14581 gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc
    14641 gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc
    14701 tgcgtgcaat ccatcttgtt caatcctagg gccgggattc tcctccacgt caccgcatgt
    14761 tagaagactt cctctgccct ctccgctgcc agatctctcg aggccctgtg ggaggaagag
    14821 aagaggtcag ataacttcgt ataatgtatg ctatacgaag ttatgcggcc gctgggagaa
    14881 gggtaagaga gctcaatcct catcttacat agaaggaagt cagtagataa tatcttacat
    14941 ttaaaagtca agaaataaca gtctgaccat actatctaga aatatggaag ttaacagtaa
    15001 ggaagttaaa acagttgaaa gtagtttcgt ctagaaagaa agacttgggg aagaacaaag
    15061 aaggctaatt ttcattatga gctttattgt aacagaaaag agaccactaa ctcagctagg
    15121 ttttcccaat tcctcaacac tttggggcac agtgtttagt cagagctgtg atcactctct
    15181 gagcacatgg attgtatctt aattttcatc tcattgaaaa tatgtaccat ttgtccaagt
    15241 ggaaactggg ctgaagcatg actgtgacca ccaccaaaga aatggctaga ctcgctaagg
    15301 tctgctgggg tgacccgtga ttgtgttgtg cttgatcttg ttgagcatgg ctcttttaag
    15361 tcaccatacc tgcccctgcc aatgcaccaa acaccaactt aaagggactt tcagcttttg
    15421 taagtcagtc tcagccttac ttctgaacca tttctttgac ttctttcatg actctgatcc
    15481 tgtcccagtt ttaagcctcc cagccaccag tcatgctgtt ttgctctgct atttacattc
    15541 ttctctacct actttggact tgatggtatg cctactatat tagttattta ttgctgtgta
    15601 acaaaccact ctgaaacata gtggcttaca accacaacca ttttatttta ttatgaaact
    15661 gtgggtcaga atgtatattg gcccattggc atgattctcc tgctattgaa aaaggaagag
    15721 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
    15781 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
    15841 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
    15901 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
    15961 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
    16021 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
    16081 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
    16141 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
    16201 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
    16261 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
    16321 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
    16381 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
    16441 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
    16501 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
    16561 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
    16621 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
    16681 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
    16741 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
    16801 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
    16861 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
    16921 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcc
//
