LOCUS       sequence-460048-       10954 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For packaging EGFP, dTomato and firefly luciferase mRNA into 
            SARS-CoV-2 virus-like particles (VLPs).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10954)
  AUTHORS   Yang Q, Davidson BA, Pajic P, Chen X, Gokcumen O, Gao M, Neelamegham
            S
  TITLE     Tuning the tropism and infectivity of SARS-CoV-2 virus-like 
            particles for mRNA delivery.
  JOURNAL   Nucleic Acids Res. 2025 Feb 27;53(5):gkaf133. doi: 
            10.1093/nar/gkaf133.
  PUBMED    40037714
REFERENCE   2  (bases 1 to 10954)
  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"; journalName: "Nucleic 
            Acids Res."; date: "2025-02-27"; volume: "53(5):gkaf133. doi"; 
            pages: " 10.1093/nar/gkaf133"
COMMENT     Sequence Label: Const.5 EGFP-P2A-dTomato-IRES-Luc-PS9
FEATURES             Location/Qualifiers
     source          1..10954
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        185..564
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        565..768
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     719..739
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     813..832
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        813..831
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      892..901
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(898..900,901..903,904..1614)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(898..900,901..903,904..1614)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(943..964)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1204..1223)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1551..1572
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1630..1686
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1693..2394
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(1826..1844)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2094..2113
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2288..2311
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    2502..2964
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2558..2575)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2785..2804
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      2985..2994
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2991..4643
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     misc_feature    5838..6426
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5891..5911)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6309..6320)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6495..6512)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6501..6725
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6771..7199
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6858..6877)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7068..7089
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7208..7228)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7213..7542
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7393..7528
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7455..7474
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             7609..8403
                     /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(7663..7682)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8273..8292
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8577..8698
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8614..8633)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8668..8687
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8747..8763)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8747..8763)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8760..8782)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8771..8787
                     /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(8795..8801,8802..8819,8820..8825))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8840..8861
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8978..8995)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9149..9737)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9229..9248)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9908..10699,10700..10768))
                     /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     10531..10550
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10769..10873)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(10948..10954,1..13))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
       61 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
      121 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
      181 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
      241 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      301 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      361 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      421 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      481 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      541 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      601 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      661 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      721 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
      781 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
      841 gctggctagc ttgttagacg aagcttgggc tgcaggtccg atccaccggt cgccaccatg
      901 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      961 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     1021 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1081 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     1141 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     1201 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1261 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1321 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     1381 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     1441 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1501 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1561 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggaagc
     1621 ggcagcggcg ccaccaactt cagcctgctg aagcaggccg gcgacgtgga ggagaacccc
     1681 ggccccgtta acatggtgag caagggcgag gaggtcatca aagagttcat gcgcttcaag
     1741 gtgcgcatgg agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc
     1801 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc
     1861 ttcgcctggg acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac
     1921 cccgccgaca tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc
     1981 gtgatgaact tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac
     2041 ggcacgctga tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta
     2101 atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc
     2161 gtgctgaagg gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg
     2221 gagttcaaga ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg
     2281 gacaccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag
     2341 cgctccgagg gccgccacca cctgttcctg tacggcatgg acgagctgta caagtaagaa
     2401 ttcgcttatg ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
     2461 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
     2521 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
     2581 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
     2641 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
     2701 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
     2761 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
     2821 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
     2881 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     2941 ttcctttgaa aaacacgatg ataaaacaca cacacaccgg tcgcgccgcc atggaagatg
     3001 ccaaaaacat taagaagggc ccagcgccat tctacccact cgaagacggg accgccggcg
     3061 agcagctgca caaagccatg aagcgctacg ccctggtgcc cggcaccatc gcctttaccg
     3121 acgcacatat cgaggtggac attacctacg ccgagtactt cgagatgagc gttcggctgg
     3181 cagaagctat gaagcgctat gggctgaata caaaccatcg gatcgtggtg tgcagcgaga
     3241 atagcttgca gttcttcatg cccgtgttgg gtgccctgtt catcggtgtg gctgtggccc
     3301 cagctaacga catctacaac gagcgcgagc tgctgaacag catgggcatc agccagccca
     3361 ccgtcgtatt cgtgagcaag aaagggctgc aaaagatcct caacgtgcaa aagaagctac
     3421 cgatcataca aaagatcatc atcatggata gcaagaccga ctaccagggc ttccaaagca
     3481 tgtacacctt cgtgacttcc catttgccac ccggcttcaa cgagtacgac ttcgtgcccg
     3541 agagcttcga ccgggacaaa accatcgccc tgatcatgaa cagtagtggc agtaccggat
     3601 tgcccaaggg cgtagcccta ccgcaccgca ccgcttgtgt ccgattcagt catgcccgcg
     3661 accccatctt cggcaaccag atcatccccg acaccgctat cctcagcgtg gtgccatttc
     3721 accacggctt cggcatgttc accacgctgg gctacttgat ctgcggcttt cgggtcgtgc
     3781 tcatgtaccg cttcgaggag gagctattct tgcgcagctt gcaagactat aagattcaat
     3841 ctgccctgct ggtgcccaca ctatttagct tcttcgctaa gagcactctc atcgacaagt
     3901 acgacctaag caacttgcac gagatcgcca gcggcggggc gccgctcagc aaggaggtag
     3961 gtgaggccgt ggccaaacgc ttccacctac caggcatccg ccagggctac ggcctgacag
     4021 aaacaaccag cgccattctg atcacccccg aaggggacga caagcctggc gcagtaggca
     4081 aggtggtgcc cttcttcgag gctaaggtgg tggacttgga caccggtaag acactgggtg
     4141 tgaaccagcg cggcgagctg tgcgtccgtg gccccatgat catgagcggc tacgttaaca
     4201 accccgaggc tacaaacgct ctcatcgaca aggacggctg gctgcacagc ggcgacatcg
     4261 cctactggga cgaggacgag cacttcttca tcgtggaccg gctgaagagc ctgatcaaat
     4321 acaagggcta ccaggtagcc ccagccgaac tggagagcat cctgctgcaa caccccaaca
     4381 tcttcgacgc cggggtcgcc ggcctgcccg acgacgatgc cggcgagctg cccgccgcag
     4441 tcgtcgtgct ggaacacggt aaaaccatga ccgagaagga gatcgtggac tatgtggcca
     4501 gccaggttac aaccgccaag aagctgcgcg gtggtgttgt gttcgtggac gaggtgccta
     4561 aaggactgac cggcaagttg gacgcccgca agatccgcga gattctcatt aaggccaaga
     4621 agggcggcaa gatcgccgtg taagaattcg cttatgcgtg cggccgcctc ctttaagaag
     4681 gagatataga tctctgtagg tcccaaacaa gctagtctta atggagtcac attaattgga
     4741 gaagccgtaa aaacacagtt caattattat aagaaagttg atggtgttgt ccaacaatta
     4801 cctgaaactt actttactca gagtagaaat ttacaagaat ttaaacccag gagtcaaatg
     4861 gaaattgatt tcttagaatt agctatggat gaattcattg aacggtataa attagaaggc
     4921 tatgccttcg aacatatcgt ttatggagat tttagtcata gtcagttagg tggtttacat
     4981 ctactgattg gactagctaa acgttttaag gaatcacctt ttgaattaga agattttatt
     5041 cctatggaca gtacagttaa aaactatttc ataacagatg cgcaaacagg ttcatctaag
     5101 tgtgtgtgtt ctgttattga tttattactt gatgattttg ttgaaataat aaaatcccaa
     5161 gatttatctg tagtttctaa ggttgtcaaa gtgactattg actatacaga aatttcattt
     5221 atgctttggt gtaaagatgg ccatgtagaa acattttacc caaaattaca atctagtcaa
     5281 gcgtggcaac cgggtgttgc tatgcctaat ctttacaaaa tgcaaagaat gctattagaa
     5341 aagtgtgacc ttcaaaatta tggtgatagt gcaacattac ctaaaggcat aatgatgaat
     5401 gtcgcaaaat atactcaact gtgtcaatat ttaaacacat taacattagc tgtaccctat
     5461 aatatgagag ttatacattt tggtgctggt tctgataaag gagttgcacc aggtacagct
     5521 gttttaagac agtggttgcc tacgggtacg ctgcttgtcg attcagatct taatgacttt
     5581 gtctctgatg cagattcaac tttgattggt gattgtgcaa ctgtacatac agctaataaa
     5641 tgggatctca ttattagtga tatgtacgac cctaagacta aaaatgttac aaaagaaaat
     5701 gactctaaag agggtttttt cacttacatt tgtgggttta tacaacaaaa gctagctctt
     5761 ggaggttccg tggctataaa gatagatccc aactccataa gtgtgtcgac atataaggat
     5821 ccactcgacg actcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     5881 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     5941 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     6001 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     6061 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     6121 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     6181 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     6241 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     6301 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     6361 ggcctcttcc gcatcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     6421 ccccgcctgg aattaattcg agctcggtac cttgattctc gagtctagag ggcccgttta
     6481 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     6541 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     6601 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     6661 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     6721 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     6781 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     6841 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     6901 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     6961 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     7021 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     7081 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     7141 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     7201 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     7261 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     7321 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     7381 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     7441 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     7501 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     7561 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     7621 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     7681 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     7741 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     7801 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     7861 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     7921 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     7981 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     8041 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     8101 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     8161 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     8221 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     8281 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     8341 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     8401 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     8461 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     8521 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     8581 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     8641 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     8701 atgtctgtat accctccacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     8761 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     8821 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     8881 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     8941 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     9001 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     9061 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     9121 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     9181 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     9241 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     9301 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     9361 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     9421 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     9481 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     9541 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     9601 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     9661 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     9721 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     9781 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     9841 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     9901 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     9961 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
    10021 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
    10081 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
    10141 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
    10201 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
    10261 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
    10321 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
    10381 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
    10441 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
    10501 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
    10561 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
    10621 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
    10681 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
    10741 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
    10801 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
    10861 taggggttcc gcgcacattt ccccgaaaag tgccacctga cctccacgga tcgggagatc
    10921 tcccgatccc ctatggtgca ctctcagtac aatc
//
