LOCUS       LPUtopia-7_CoV-2        9911 bp DNA     circular SYN 19-SEP-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9911)
  AUTHORS   Gabor Balazsi
  TITLE     Direct Submission
  JOURNAL   Exported Sep 20, 2024 from SnapGene 7.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1..40)
                     /label=pUtopia-7-CoV-2 target-2.HiR
     primer_bind     1..40
                     /label=pUtopia-7-CoV-2 target-1.HiF
     primer_bind     complement(1..20)
                     /label=pUtopia-7-CoV1 target-1.HiR
     promoter        complement(20)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_feature    21..789
                     /locus_tag="RdRp region"
                     /label=RdRp region
     misc_feature    81..102
                     /label=CrCOV 20
     misc_feature    790..2042
                     /locus_tag="N gene"
                     /label=N gene
     misc_feature    834..855
                     /label=CrCOV 21
     misc_feature    980..1001
                     /label=CrCOV 24
     primer_bind     2064..2086
                     /label=1167_seq
     enhancer        2073..2664
                     /locus_tag="WPRE"
     primer_bind     2662..2701
                     /label=pUtopia-7-CoV-1-2 target-2.HiF
     primer_bind     complement(2662..2701)
                     /label=pUtopia-7-CoV-1-2 target-1.HiR
     promoter        complement(2682..2684)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2684..2703)
                     /label=RIME-1.4
     primer_bind     2684..2703
                     /label=RIME-1.2
     polyA_signal    2686..2836
     primer_bind     2704..2725
                     /label=bGH-Blst.F
     primer_bind     2704..2725
                     /label=bGH-Blst(2).F
     primer_bind     2704..2722
                     /label=bGH-Blst(4).F
     primer_bind     2704..2722
                     /label=bGH-Blst(3).F
     primer_bind     2709..2727
                     /label=pU3-NeoR.F
     polyA_signal    2710..2836
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2710..2730
                     /label=pUtopia53.HiR
     primer_bind     2731..2754
                     /label=TG1.6.9
     primer_bind     complement(2767..2781)
                     /label=AAVS1.DxR
                     /note="AAVS1 diagnostic PCR, forward. PCR product 394bp for
                     wild-type (NC_000019.10). Products on potentially 
                     unintended templates:>NC_000001.11 Homo sapiens chromosome 
                     1, GRCh38.p7 Primary Assemblyproduct length = 2209Features 
                     associated with this product:  T-cell surface glycoprotein 
                     CD3 zeta chain isoform 2 prec...  T-cell surface 
                     glycoprotein CD3 zeta chain isoform 1 prec... product 
                     length = 3051Features associated with this product: group 
                     IIF secretory phospholipase A2 isoform X1 group IIF 
                     secretory phospholipase A2>NC_000004.12 Homo sapiens 
                     chromosome 4, GRCh38.p7 Primary Assembly product length = 
                     3333 Features associated with this product: calcium 
                     uniporter regulatory subunit MCUb, mitochondrial calcium 
                     uniporter regulatory subunit MCUb, mitochondrial 
                     >NC_000007.14 Homo sapiens chromosome 7, GRCh38.p7 Primary 
                     Assembly  product length = 1258 Features flanking this 
                     product: 104529 bp at 5' side: histone-lysine 
                     N-methyltransferase EZH2 isoform X22 47538 bp at 3' side: 
                     protein disulfide-isomerase A4 precursor"
     polyA_signal    2837..2934
                     /label=bGH poly(A)
     primer_bind     complement(2914..2934)
                     /label=bGH-pA-r
     primer_bind     complement(2918..2934)
                     /label=RFP-AAVS1.HiR
     primer_bind     2923..2942
                     /label=protrap_Fluo-AAVS1_ext.HiF v2
     primer_bind     complement(2923..2942)
                     /label=pUt7.0M-fluo-red.HiF
     primer_bind     complement(2923..2942)
                     /label=protrap_Fluo-AAVS1_RFP.HiR
     primer_bind     2923..2942
                     /label=protrap_Fluo-AAVS1_ext.HiR
     primer_bind     3047..3076
                     /label=Put7.0_Intermediate_Ext_HiF
     primer_bind     3055..3074
                     /label=protrap_Fluo-AAVS1_ext.HiR
     primer_bind     3063..3094
                     /label=FRT-Ori-OL-f
     rep_origin      complement(3095..3683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3182..3201)
                     /label=pU3_pciI_seq_R
     primer_bind     complement(3245..3274)
                     /label=HiFiOriSplit.Rev
     primer_bind     3245..3274
                     /label=HifiOriSplit.Fwd
     primer_bind     complement(3473..3502)
                     /label=Ori-Split.Rev
     primer_bind     3473..3502
                     /label=Ori-Split.fwd
     primer_bind     3675..3696
                     /label=nvn.1.18
     primer_bind     complement(3684..3708)
                     /label=Ori-OL-r
     primer_bind     complement(3690..3708)
                     /label=Blst-FRT-Ori.R
     primer_bind     3832..3853
                     /label=nvn.1.18
     primer_bind     complement(3832..3851)
                     /label=Int_HiR
     primer_bind     3832..3851
                     /label=Ext_HiF
     CDS             complement(3854..4714)
                     /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     4217..4236
                     /label=AmpR split fwd
     primer_bind     complement(4217..4236)
                     /label=AmpR-split.rev
                     /note="Primer to split AmpR for HiFi assembly"
     promoter        complement(4715..4819)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4751..4770
                     /label=AmpPromoter-split.rev
     primer_bind     complement(4751..4770)
                     /label=AmpPromoter-split.fwd
     primer_bind     complement(4822..4841)
                     /label=nvn.1.17
     rep_origin      4845..5300
                     /direction=RIGHT
                     /label=F1 ori
     CDS             complement(5302..5370)
                     /label=LacZ alpha
     primer_bind     5442..5458
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5826..6056
                     /label=core insulin
     primer_bind     5868..5877
                     /label=TG1.6.9
     primer_bind     complement(6046..6064)
                     /label=protrap_Fluo-AAVS1_ext.HiF v2
     primer_bind     6046..6064
                     /label=protrap_Fluo-AAVS1_RFP.HiR
     primer_bind     6070..6088
                     /label=protrap_Fluo-AAVS1_int.HiF v2
     primer_bind     complement(6070..6088)
                     /label=protrap_Fluo-AAVS1_RFP.HiFa
     misc_feature    6113..6916
                     /label=AAVS1-HA-L
     primer_bind     complement(6664..6674)
                     /label=KRASB-HSV.F
     primer_bind     6714..6733
                     /label=AAVS1.DxR
                     /note="AAVS1 diagnostic PCR, forward. PCR product 394bp for
                     wild-type (NC_000019.10). Products on potentially 
                     unintended templates:>NC_000001.11 Homo sapiens chromosome 
                     1, GRCh38.p7 Primary Assemblyproduct length = 2209Features 
                     associated with this product:  T-cell surface glycoprotein 
                     CD3 zeta chain isoform 2 prec...  T-cell surface 
                     glycoprotein CD3 zeta chain isoform 1 prec... product 
                     length = 3051Features associated with this product: group 
                     IIF secretory phospholipase A2 isoform X1 group IIF 
                     secretory phospholipase A2>NC_000004.12 Homo sapiens 
                     chromosome 4, GRCh38.p7 Primary Assembly product length = 
                     3333 Features associated with this product: calcium 
                     uniporter regulatory subunit MCUb, mitochondrial calcium 
                     uniporter regulatory subunit MCUb, mitochondrial 
                     >NC_000007.14 Homo sapiens chromosome 7, GRCh38.p7 Primary 
                     Assembly  product length = 1258 Features flanking this 
                     product: 104529 bp at 5' side: histone-lysine 
                     N-methyltransferase EZH2 isoform X22 47538 bp at 3' side: 
                     protein disulfide-isomerase A4 precursor"
     primer_bind     6748..6769
                     /label=AAVS1.DxRv2
     primer_bind     6808..6827
                     /label=pU3_AAVS1_seq_F
     primer_bind     6833..6856
                     /label=pU53_AAVS1_F-1.1
     primer_bind     complement(6885..6906)
                     /label=protrap_Fluo-AAVS1_ext.HiF
     primer_bind     complement(6899..6916)
                     /label=AAVS1-HA-Left-C-Term-Seq-r
     primer_bind     6899..6916
                     /label=AAVS1-HA-Left-C-Term-Seq-f
     primer_bind     6921..6940
                     /label=protrap_Fluo-AAVS1_int.HiF
     primer_bind     complement(6922..6941)
                     /label=Ext_HiR
     primer_bind     6922..6941
                     /label=FRT_HiF
     misc_binding    6945..6992
     protein_bind    6945..6992
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_recomb     6945..6992
                     /label=FRT Prime
     primer_bind     6945..6992
                     /label=mLin-BACH1.F
     primer_bind     6945..6992
                     /label=mLin-BACH1-SGC.F
     primer_bind     6946..6972
                     /label=FRT-Ori-OL-f
     primer_bind     complement(6999..7017)
                     /label=pUtopia53.HiF_v2
     protein_bind    7013..7046
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     complement(7106..7125)
                     /label=AAVS1-pUt-head.seq
     misc_feature    7122..7330
                     /label=Geneious type: promoter uukaryotic
                     /note="Geneious type: promoter uukaryotic"
     primer_bind     complement(7177..7196)
                     /label=AAVS1-pUt-head.seq
     primer_bind     complement(7315..7333)
                     /label=Vector.FOR
     CDS             7331..8134
     CDS             7331..8134
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     7331..7350
                     /label=RIME-1.3
     primer_bind     complement(7332..7359)
                     /label=pUtopia 5.3 head internal primerv2
     primer_bind     7332..7359
                     /label=pUtopia 5.3 head internal primer
     primer_bind     complement(7345..7364)
                     /label=pUt7.0M-NeoR-split.HiR
     primer_bind     7345..7364
                     /label=pUt7.0M-NeoR-split.HiF
     primer_bind     7349..7368
                     /label=NeoR KanR Fwd
     primer_bind     complement(7394..7413)
                     /label=NeoR-KanR rev
     primer_bind     7463..7484
                     /label=KasI-NeoR-f
     primer_bind     7467..7476
                     /label=KRAS4B.HiR
     primer_bind     complement(7473..7492)
                     /label=Put7.0_Intermediate_Ext_HiR
     primer_bind     7473..7492
                     /label=Put7.0_Intermediate_Int_HiF
     primer_bind     complement(8105..8131)
                     /label=Primer 1
     primer_bind     complement(8113..8131)
                     /label=TGKA_pu42_rev
     primer_bind     complement(8119..8138)
                     /label=RIME-1.4
     primer_bind     8119..8138
                     /label=RIME-1.2
     primer_bind     8131..8154
                     /label=Vector.REV
     primer_bind     8133..8153
                     /label=TGKA_pu42_fwd
     primer_bind     8136..8170
                     /label=TGKA_pu42_fwd
     polyA_signal    8141..8189
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(8170..8189)
                     /label=HSV_TK_rev
                     /note="3'Tm=55.8 3'Ta(annealing temp)=55.8"
     primer_bind     complement(8171..8190)
                     /label=RIME-1.4-2
     primer_bind     complement(8171..8189)
                     /label=HSV_TK-FRT-Rev
     primer_bind     complement(8172..8192)
                     /label=HSV-TK-FRT'_rev
     protein_bind    8200..8233
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     complement(8379..8403)
                     /label=pU53 amp pu42 vector REV
     primer_bind     8447..8462
                     /label=pu52 amp pu42 FWD
     primer_bind     8452..8468
                     /label=pu53 amp pu42 vector FOR
     primer_bind     complement(8453..8470)
                     /label=pU3-NeoR.R
     primer_bind     8465..8498
                     /label=CMV-SgrDI-mut-f
     primer_bind     complement(8465..8483)
                     /label=pU3_NeoR_seq_R
     misc_feature    8467..9121
                     /label=Geneious type: promoter uukaryotic
                     /note="Geneious type: promoter uukaryotic"
     primer_bind     8473..8490
                     /label=mLin-BACH1.F
     primer_bind     8473..8490
                     /label=mLin-BACH1-SGC.F
     primer_bind     8485..8552
                     /label=MluI-CMV
     primer_bind     complement(8488..8507)
                     /label=GFP_Linearizer_FRT.HiR
     primer_bind     8490..8528
                     /label=CMV-D2i-SgrDI-f
     primer_bind     8490..8516
                     /label=NsiI-D2ir-f
     enhancer        8493..8872
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     8493..8533
                     /label=SalI-CMV-D2ir-f
     primer_bind     8493..8518
                     /label=CMV-enhancer-upstream-seq
     primer_bind     8796..8815
                     /label=Put7.0_Intermediate_RFP_HiF
     primer_bind     complement(8817..8839)
                     /label=protrap_Fluo-AAVS1_int.HiF v2
     primer_bind     8817..8839
                     /label=protrap_Fluo-AAVS1_RFP.HiFa
     primer_bind     8820..8839
                     /label=pUt7.0M-fluo-red.HiR
     primer_bind     8820..8839
                     /label=protrap_Fluo-AAVS1_RFP.HiF
     primer_bind     complement(8829..8857)
                     /label=pAAVS1_rev(3)
     primer_bind     complement(8829..8857)
                     /label=pAAVS1-Rev(2)
     primer_bind     8838..8890
                     /label=FRT-CMV_Promoter-f
     primer_bind     complement(8838..8872)
                     /label=HR(AAVS1)-RFP.HiR
     promoter        8873..9076
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8873..8893
                     /label=CMV_Promoter_fwd
                     /note="3'Tm=56.1 3'Ta(annealing temp)=55.0"
     primer_bind     8887..8906
                     /label=CMV-seq.fwd
                     /note="Sequencing primer from CMV"
     primer_bind     complement(9052..9076)
                     /label=CMV_Promoter_rev
                     /note="3'Tm=55.0 3'Ta(annealing temp)=55.0"
     TATA_signal     9062..9068
     promoter        9121..9139
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9171..9186)
                     /label=KRAS4B.HiR
     CDS             9184..9903
     CDS             join(9184..9186,9187..9189,9190..9903)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     9184..9205
                     /label=hEGFP-f
     primer_bind     9184..9204
                     /label=2A-NeonGreen.F
     primer_bind     complement(9184..9202)
                     /label=hEGFP-N-Term-Seq-r
     primer_bind     9184..9201
                     /label=HiFi-KRAS-GFP.fwd
     primer_bind     9184..9198
                     /label=pUt-NF-GFP_NVN_no-Bach1_HiFi-F
     primer_bind     9185..9204
                     /label=TG1.10.5
     primer_bind     9283..9302
                     /label=EGFP sgRNA target
     primer_bind     9296..9314
                     /label=EGFP37.fwd
     primer_bind     complement(9296..9313)
                     /label=EGFP43.rev
     primer_bind     9491..9511
                     /label=TG1.10.3
     primer_bind     complement(9516..9533)
                     /label=hEGFP-Midstream-Middle-Seq-r
     primer_bind     9829..9847
                     /label=hEGFP-C-Term-Seq-f
     primer_bind     9833..9852
                     /label=pUtopia 5.3 tail internal primerv3
     primer_bind     9876..9903
                     /label=pUtopia 5.3 tail internal primerv2
     primer_bind     complement(9876..9903)
                     /label=pUtopia 5.3 tail internal primer
     primer_bind     complement(9879..9900)
                     /label=pUt-NF-GFP_NVN_Mutagenesis-R
     primer_bind     complement(9879..9900)
                     /label=mLin-Vec.R
     primer_bind     complement(9879..9900)
                     /label=T2a-hEGFP-OL-r
     primer_bind     complement(9880..9908)
                     /label=hEGFP-AgeI-BGH-pA-1stOL-r
     primer_bind     complement(9880..9899)
                     /label=EGFP Fwd SL
     primer_bind     complement(9881..9900)
                     /label=G-Linker-OL-r
     primer_bind     complement(9889..9903)
                     /label=BamHI-hEGFP-r
     primer_bind     complement(9891..9908)
                     /label=AgeI-hEGFP-r
ORIGIN
        1 cgactctaga gggccctatt atggtcatgt gtggcggttc actatatgtt aaaccaggtg
       61 gaacctcatc aggagatgcc acaactgctt atgctaatag tgtttttaac atttgtcaag
      121 ctgtcacggc caatgttaat gcacttttat ctactgatgg taacaaaatt gccgataagt
      181 atgtccgcaa tttacaacac agactttatg agtgtctcta tagaaataga gatgttgaca
      241 cagactttgt gaatgagttt tacgcatatt tgcgtaaaca tttctcaatg atgatactct
      301 ctgacgatgc tgttgtgtgt ttcaatagca cttatgcatc tcaaggtcta gtggctagca
      361 taaagaactt taagtcagtt ctttattatc aaaacaatgt ttttatgtct gaagcaaaat
      421 gttggactga gactgacctt actaaaggac ctcatgaatt ttgctctcaa catacaatgc
      481 tagttaaaca gggtgatgat tatgtgtacc ttccttaccc agatccatca agaatcctag
      541 gggccggctg ttttgtagat gatatcgtaa aaacagatgg tacacttatg attgaacggt
      601 tcgtgtcttt agctatagat gcttacccac ttactaaaca tcctaatcag gagtatgctg
      661 atgtctttca tttgtactta caatacataa gaaagctaca tgatgagtta acaggacaca
      721 tgttagacat gtattctgtt atgcttacta atgataacac ttcaaggtat tgggaacctg
      781 agttttatgt ctgataatgg ccccaaaatc agcgaaatgc accccgcatt acgtttggtg
      841 gaccctcaga ttcaactggc agtaaccaga atggagaacg cagtggggcg cgatcaaaac
      901 aacgtcggcc ccaaggttta cccaataata ctgcgtcttg gttcaccgct ctcactcaac
      961 atggcaagga agaccttaaa ttccctcgag gacaaggcgt tccaattaac accaatagca
     1021 gtccagatga ccaaattggc tactaccgaa gagctaccag acgaattcgt ggtggtgacg
     1081 gtaaaatgaa agatctcagt ccaagatggt atttctacta cctaggaact gggccagaag
     1141 ctggacttcc ctatggtgct aacaaagacg gcatcatatg ggttgcaact gagggagcct
     1201 tgaatacacc aaaagatcac attggcaccc gcaatcctgc taacaatgct gcaatcgtgc
     1261 tacaacttcc tcaaggaaca acattgccaa aaggcttcta cgcagaaggg agcagaggcg
     1321 gcagtcaagc ctcttctcgt tcctcatcac gtagtcgcaa cagttcaaga aattcaactc
     1381 caggcagcag taggggaact tctcctgcta gaatggctgg caatggcggt gatgctgctc
     1441 ttgctttgct gctgcttgac agattgaacc agcttgagag caaaatgtct ggtaaaggcc
     1501 aacaacaaca aggccaaact gtcactaaga aatctgctgc tgaggcttct aagaagcctc
     1561 ggcaaaaacg tactgccact aaagcataca atgtaacaca agctttcggc agacgtggtc
     1621 cagaacaaac ccaaggaaat tttggggacc aggaactaat cagacaagga actgattaca
     1681 aacattggcc gcaaattgca caatttgccc ccagcgcttc agcgttcttc ggaatgtcgc
     1741 gcattggcat ggaagtcaca ccttcgggaa cgtggttgac ctacacaggt gccatcaaat
     1801 tggatgacaa agatccaaat ttcaaagatc aagtcatttt gctgaataag catattgacg
     1861 catacaaaac attcccacca acagagccta aaaaggacaa aaagaagaag gctgatgaaa
     1921 ctcaagcctt accgcagaga cagaagaaac agcaaactgt gactcttctt cctgctgcag
     1981 atttggatga tttctccaaa caattgcaac aatccatgag cagtgctgac tcaactcagg
     2041 ccggcatgca agcttgatat caagcttatc gataatcaac ctctggatta caaaatttgt
     2101 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     2161 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2221 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     2281 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     2341 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     2401 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     2461 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     2521 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     2581 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     2641 tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gaatgctaga gctcgctgat
     2701 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2761 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2821 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2881 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggagatct
     2941 gcggccgcga aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc
     3001 cacagtcccc gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaaagg
     3061 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3121 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3181 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3241 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3301 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3361 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3421 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3481 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3541 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3601 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3661 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3721 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3781 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3841 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3901 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3961 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4021 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4081 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4141 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4201 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4261 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4321 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4381 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4441 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4501 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4561 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4621 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4681 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4741 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4801 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
     4861 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4921 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     4981 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     5041 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     5101 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     5161 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     5221 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     5281 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
     5341 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     5401 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     5461 gcgcgcctcg ttcattcacg tttttgaacc cgtggaggac gggcagactc gcggtgcaaa
     5521 tgtgttttac agcgtgatgg agcagatgaa gatgctcgac acgctgcaga acacgcagct
     5581 agattaaccc tagaaagata atcatattgt gacgtacgtt aaagataatc atgtgtaaaa
     5641 ttgacgcatg tgttttatcg gtctgtatat cgaggtttat ttattaattt gaatagatat
     5701 taagttttat tatatttaca cttacatact aataataaat tcaacaaaca atttatttat
     5761 gtttatttat ttattaaaaa aaacaaaaac tcaaaatttc ttctataaag taacaaaact
     5821 tttatgaggg acagcccccc ccaaagcccc cagggatgta attacgtccc tcccccgcta
     5881 gggggcagca gcgagccgcc cggggctccg ctccggtccg gcgctccccc cgcatccccg
     5941 agccggcagc gtgcggggac agcccgggca cggggaaggt ggcacgggat cgctttcctc
     6001 tgaacgcttc tcgctgctct ttgagcctgc agacacctgg ggggatacgg ggaaaaggcc
     6061 tccacggcca ctagtaacgg ccgccagtgt gctggaattc gcccttggta cctgctttct
     6121 ctgaccagca ttctctcccc tgggcctgtg ccgctttctg tctgcagctt gtggcctggg
     6181 tcacctctac ggctggccca gatccttccc tgccgcctcc ttcaggttcc gtcttcctcc
     6241 actccctctt ccccttgctc tctgctgtgt tgctgcccaa ggatgctctt tccggagcac
     6301 ttccttctcg gcgctgcacc acgtgatgtc ctctgagcgg atcctccccg tgtctgggtc
     6361 ctctccgggc atctctcctc cctcacccaa ccccatgccg tcttcactcg ctgggttccc
     6421 ttttccttct ccttctgggg cctgtgccat ctctcgtttc ttaggatggc cttctccgac
     6481 ggatgtctcc cttgcgtccc gcctcccctt cttgtaggcc tgcatcatca ccgtttttct
     6541 ggacaacccc aaagtacccc gtctccctgg ctttagccac ctctccatcc tcttgctttc
     6601 tttgcctgga caccccgttc tcctgtggat tcgggtcacc tctcactcct ttcatttggg
     6661 cagctcccct acccccctta cctctctagt ctgtgctagc tcttccagcc ccctgtcatg
     6721 gcatcttcca ggggtccgag agctcagcta gtcttcttcc tccaacccgg gcccctatgt
     6781 ccacttcagg acagcatgtt tgctgcctcc agggatcctg tgtccccgag ctgggaccac
     6841 cttatattcc cagggccggt taatgtggct ctggttctgg gtacttttat ctgtcccctc
     6901 caccccacag tggggcacgc gtaagggcga attctgcaga tatcgaagtt cctattccga
     6961 agttcctatt cttcaaatag tataggaact tcgtcgagat cgatggcgcg ccataacttc
     7021 gtatagcata cattatacga agttatgaat ttcgagcagt gtggttttca agaggaagca
     7081 aaaagcctct ccacccaggc ctggaatgtt tccacccaat gtcgagcagt gtggttttgc
     7141 aagaggaagc aaaaagcctc tccacccagg cctggaatgt ttccacccaa tgtcgagcaa
     7201 accccgccca gcgtcttgtc attggcgaat tcgaacacgc agatgcagtc ggggcggcgc
     7261 ggtcccaggt ccacttcgca tattaaggtg acgcgtgtgg cctcgaacac cgagcgaccc
     7321 tgcagccaat atgggatcgg ccattgaaca agatggattg cacgcaggtt ctccggccgc
     7381 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     7441 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     7501 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     7561 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     7621 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     7681 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     7741 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     7801 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     7861 caaggcgcgc atgcccgacg gcgatgatct cgtcgtgacc catggcgatg cctgcttgcc
     7921 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     7981 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     8041 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     8101 cgccttctat cgccttcttg acgagttctt ctgaggggat cggcaataaa aagacagaat
     8161 aaaacgcacg ggtgttgggt cgtttgttcg gatcaattca taacttcgta tagcatacat
     8221 tatacgaagt tatggcgcgc cttaattaag gatccgtcga cggatcggga gatctcccga
     8281 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     8341 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     8401 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     8461 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
     8521 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     8581 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     8641 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     8701 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     8761 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     8821 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     8881 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     8941 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     9001 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     9061 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
     9121 taatacgact cactataggg agacccaagc ttggtaccga gctcggatcc accggtcgcc
     9181 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     9241 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     9301 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     9361 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     9421 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     9481 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     9541 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     9601 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     9661 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     9721 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     9781 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     9841 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     9901 taaagcggcc g
//
