LOCUS       sequence-493694-        9787 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IVT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9787)
  AUTHORS   Datlinger P, Pankevich EV, Arnold CD, Pranckevicius N, Lin J, 
            Romanovskaia D, Schaefer M, Piras F, Orts AC, Nemc A, Biesaga PN, 
            Chan M, Neuwirth T, Artemov AV, Li W, Ladstatter S, Krausgruber T, 
            Bock C
  TITLE     Systematic discovery of CRISPR-boosted CAR T cell immunotherapies.
  JOURNAL   Nature. 2025 Oct;646(8086):963-972. doi: 10.1038/s41586-025-09507-9.
            Epub 2025 Sep 24.
  PUBMED    40993398
REFERENCE   2  (bases 1 to 9787)
  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/s41586-025-09507-9"; journalName: "Nature"; date: 
            "2025-10"; volume: "646"; issue: "8086"; pages: "963-972"
COMMENT     Sequence Label: pEP0004_pIVTRup(BsmBI)_AncBE4max
FEATURES             Location/Qualifiers
     source          1..9787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(277..293)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(277..293)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(290..312)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    301..317
                     /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(325..331,332..349,350..355))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    370..391
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(508..525)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(679..1267)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(759..778)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1438..2229,2230..2298))
                     /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     2061..2080
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(2316..3110)
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2427..2446)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3037..3056
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      complement(3454..3882)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3564..3585)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3776..3795
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4008..4030
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4022..4039
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4023..4039
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4046..4064
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        4136..4154
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4236..4256
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(5037..5060,5061..5063,5064..9137)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5037..9137
                     /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
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             9168..9416
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             9447..9695
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             9738..9758
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 tctggccatg cccttcttct ctcccttgca cctgtacctc ttggtctttg aataaagcct
       61 gagtaggaag tgagggtcta gaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
      121 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaatgaga cgcgatcaat aaccgacact
      181 cctgcgtcgc ctacgcgcta cgatctgcag aattccagca cactggcggc cgttactagt
      241 ggatccgagc tcggtaccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
      301 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
      361 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
      421 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
      481 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
      541 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      601 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      661 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      721 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      781 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      841 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      901 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      961 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1021 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1081 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1141 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1201 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1261 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1321 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1381 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1441 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1501 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1561 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1621 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1681 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1741 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1801 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1861 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1921 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1981 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2041 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2101 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2161 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2221 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2281 gaaatgttga atactcatac tcttcctttt tcaattcaga agaactcgtc aagaaggcga
     2341 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
     2401 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
     2461 cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
     2521 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
     2581 cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
     2641 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
     2701 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     2761 gccatgatgg atactttctc ggcaggagca aggtgggatg acaggagatc ctgccccggc
     2821 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
     2881 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg cagttcattc
     2941 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
     3001 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     3061 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
     3121 cctcatcctg tctcttgatc agatcttgat cccctgcgcc atcagatcct tggcggcaag
     3181 aaagccatcc agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc
     3241 aattccggtt cgcttgctgt ccataaaacc gcccagtcta gctatcgcca tgtaagccca
     3301 ctgcaagcta cctgctttct ctttgcgctt gcgttttccc ttgtccagat agcccagtag
     3361 ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctacgt gttccgcttc
     3421 ctttagcagc ccttgcgccc tgaattttgt taaaattcgc gttaaatttt tgttaaatca
     3481 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3541 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3601 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3661 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3721 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3781 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     3841 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct
     3901 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     3961 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     4021 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tgggccctct
     4081 agatgcatgc tcgagcggcc gccagtgtga tggatttgga ccctcgtaca gaagctaata
     4141 cgactcacta tagggaaata agagagaaaa gaagagtaag aagaaatata agagccacca
     4201 tgaaacggac agccgacgga agcgagttcg agtcaccaaa gaagaagcgg aaagtcagca
     4261 gtgaaaccgg accagtggca gtggacccaa ccctgaggag acggattgag ccccatgaat
     4321 ttgaagtgtt ctttgaccca agggagctga ggaaggagac atgcctgctg tacgagatca
     4381 agtggggcac aagccacaag atctggcgcc acagctccaa gaacaccaca aagcacgtgg
     4441 aagtgaattt catcgagaag tttacctccg agcggcactt ctgcccctct accagctgtt
     4501 ccatcacatg gtttctgtct tggagccctt gcggcgagtg ttccaaggcc atcaccgagt
     4561 tcctgtctca gcaccctaac gtgaccctgg tcatctacgt ggcccggctg tatcaccaca
     4621 tggaccagca gaacaggcag ggcctgcgcg atctggtgaa ttctggcgtg accatccaga
     4681 tcatgacagc cccagagtac gactattgct ggcggaactt cgtgaattat ccacctggca
     4741 aggaggcaca ctggccaaga tacccacccc tgtggatgaa gctgtatgca ctggagctgc
     4801 acgcaggaat cctgggcctg cctccatgtc tgaatatcct gcggagaaag cagccccagc
     4861 tgacattttt caccattgct ctgcagtctt gtcactatca gcggctgcct cctcatattc
     4921 tgtgggctac aggcctgaag tctggaggat ctagcggagg atcctctggc agcgagacac
     4981 caggaacaag cgagtcagca acaccagaga gcagtggcgg cagcagcggc ggcagcgaca
     5041 agaagtacag catcggcctg gccatcggca ccaactctgt gggctgggcc gtgatcaccg
     5101 acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca
     5161 tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc
     5221 ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc
     5281 aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag
     5341 agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg
     5401 tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg
     5461 tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca
     5521 agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca
     5581 agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg
     5641 ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg
     5701 aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg
     5761 ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca
     5821 aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg
     5881 gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga
     5941 gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca
     6001 agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc
     6061 tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca
     6121 ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga
     6181 tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc
     6241 ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg cacgccattc
     6301 tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga
     6361 tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg
     6421 cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg
     6481 acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc
     6541 ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg
     6601 agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg
     6661 agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc
     6721 agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg
     6781 tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg
     6841 acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc
     6901 tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt
     6961 tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga
     7021 gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc
     7081 tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga
     7141 cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc
     7201 acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg
     7261 tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa
     7321 tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc
     7381 ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa
     7441 acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt
     7501 acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc
     7561 ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga
     7621 accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact
     7681 ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg
     7741 ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg
     7801 aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt
     7861 acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg
     7921 tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc
     7981 acgcccacga cgcctaccta aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta
     8041 agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg
     8101 ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca
     8161 tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga
     8221 tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg
     8281 tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag
     8341 gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa
     8401 agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg
     8461 tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc
     8521 tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg
     8581 aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc
     8641 tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg
     8701 gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg
     8761 agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca
     8821 agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg
     8881 ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca
     8941 gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg
     9001 ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc
     9061 tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt
     9121 ctcagctggg aggtgacagc ggcgggagcg gcgggagcgg ggggagcact aatctgagcg
     9181 acatcattga gaaggagact gggaaacagc tggtcattca ggagtccatc ctgatgctgc
     9241 ctgaggaggt ggaggaagtg atcggcaaca agccagagtc tgacatcctg gtgcacaccg
     9301 cctacgacga gtccacagat gagaatgtga tgctgctgac ctctgacgcc cccgagtata
     9361 agccttgggc cctggtcatc caggattcta acggcgagaa taagatcaag atgctgagcg
     9421 gaggatccgg aggatctgga ggcagcacca acctgtctga catcatcgag aaggagacag
     9481 gcaagcagct ggtcatccag gagagcatcc tgatgctgcc cgaagaagtc gaagaagtga
     9541 tcggaaacaa gcctgagagc gatatcctgg tccataccgc ctacgacgag agtaccgacg
     9601 aaaatgtgat gctgctgaca tccgacgccc cagagtataa gccctgggct ctggtcatcc
     9661 aggattccaa cggagagaac aaaatcaaaa tgctgtctgg cggctcaaaa agaaccgccg
     9721 acggcagcga attcgagccc aagaagaaga ggaaagtctg aatcgctgcc ttctgcgggg
     9781 cttgcct
//
