LOCUS       sequence-462178-       11728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For packaging EGFP, dTomato, TagBFP 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 11728)
  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 11728)
  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.6 EGFP-P2A-dTomato-T2A-TagBFP-IRES-Luc-PS9
FEATURES             Location/Qualifiers
     source          1..11728
                     /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"
     CDS             2413..2466
                     /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"
     CDS             2467..3168
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    3276..3738
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3332..3349)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3559..3578
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      3759..3768
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3765..5417
                     /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    6612..7200
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6665..6685)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7083..7094)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7269..7286)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7275..7499
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7545..7973
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7632..7651)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7842..7863
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7982..8002)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7987..8316
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8167..8302
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8229..8248
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             8383..9177
                     /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(8437..8456)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9047..9066
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    9351..9472
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9388..9407)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9442..9461
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9521..9537)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9521..9537)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9534..9556)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9545..9561
                     /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(9569..9575,9576..9593,9594..9599))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9614..9635
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9752..9769)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9923..10511)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10003..10022)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10682..11473,11474..11542))
                     /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     11305..11324
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11543..11647)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(11722..11728,1..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 caagggcggc
     2401 gggtccggag gagagggcag aggaagtctc ctaacatgcg gtgacgtgga ggagaatcct
     2461 ggcccaatga gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc
     2521 gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
     2581 cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
     2641 gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg catccccgac
     2701 ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaggac
     2761 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
     2821 gtcaagatca gaggggtgaa cttcacatcc aacggccctg tgatgcagaa gaaaacactc
     2881 ggctgggagg ccttcaccga gacgctgtac cccgctgacg gcggcctgga aggcagaaac
     2941 gacatggccc tgaagctcgt gggcgggagc catctgatcg caaacatcaa gaccacatat
     3001 agatccaaga aacccgctaa gaacctcaag atgcctggcg tctactatgt ggactacaga
     3061 ctggaaagaa tcaaggaggc caacaacgag acctacgtcg agcagcacga ggtggcagtg
     3121 gccagatact gcgacctccc tagcaaactg gggcacaagc ttaattgatg cgaattcgct
     3181 tatgctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     3241 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     3301 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
     3361 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     3421 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     3481 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     3541 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     3601 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt
     3661 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     3721 tgaaaaacac gatgataaaa cacacacaca ccggtcgcgc cgccatggaa gatgccaaaa
     3781 acattaagaa gggcccagcg ccattctacc cactcgaaga cgggaccgcc ggcgagcagc
     3841 tgcacaaagc catgaagcgc tacgccctgg tgcccggcac catcgccttt accgacgcac
     3901 atatcgaggt ggacattacc tacgccgagt acttcgagat gagcgttcgg ctggcagaag
     3961 ctatgaagcg ctatgggctg aatacaaacc atcggatcgt ggtgtgcagc gagaatagct
     4021 tgcagttctt catgcccgtg ttgggtgccc tgttcatcgg tgtggctgtg gccccagcta
     4081 acgacatcta caacgagcgc gagctgctga acagcatggg catcagccag cccaccgtcg
     4141 tattcgtgag caagaaaggg ctgcaaaaga tcctcaacgt gcaaaagaag ctaccgatca
     4201 tacaaaagat catcatcatg gatagcaaga ccgactacca gggcttccaa agcatgtaca
     4261 ccttcgtgac ttcccatttg ccacccggct tcaacgagta cgacttcgtg cccgagagct
     4321 tcgaccggga caaaaccatc gccctgatca tgaacagtag tggcagtacc ggattgccca
     4381 agggcgtagc cctaccgcac cgcaccgctt gtgtccgatt cagtcatgcc cgcgacccca
     4441 tcttcggcaa ccagatcatc cccgacaccg ctatcctcag cgtggtgcca tttcaccacg
     4501 gcttcggcat gttcaccacg ctgggctact tgatctgcgg ctttcgggtc gtgctcatgt
     4561 accgcttcga ggaggagcta ttcttgcgca gcttgcaaga ctataagatt caatctgccc
     4621 tgctggtgcc cacactattt agcttcttcg ctaagagcac tctcatcgac aagtacgacc
     4681 taagcaactt gcacgagatc gccagcggcg gggcgccgct cagcaaggag gtaggtgagg
     4741 ccgtggccaa acgcttccac ctaccaggca tccgccaggg ctacggcctg acagaaacaa
     4801 ccagcgccat tctgatcacc cccgaagggg acgacaagcc tggcgcagta ggcaaggtgg
     4861 tgcccttctt cgaggctaag gtggtggact tggacaccgg taagacactg ggtgtgaacc
     4921 agcgcggcga gctgtgcgtc cgtggcccca tgatcatgag cggctacgtt aacaaccccg
     4981 aggctacaaa cgctctcatc gacaaggacg gctggctgca cagcggcgac atcgcctact
     5041 gggacgagga cgagcacttc ttcatcgtgg accggctgaa gagcctgatc aaatacaagg
     5101 gctaccaggt agccccagcc gaactggaga gcatcctgct gcaacacccc aacatcttcg
     5161 acgccggggt cgccggcctg cccgacgacg atgccggcga gctgcccgcc gcagtcgtcg
     5221 tgctggaaca cggtaaaacc atgaccgaga aggagatcgt ggactatgtg gccagccagg
     5281 ttacaaccgc caagaagctg cgcggtggtg ttgtgttcgt ggacgaggtg cctaaaggac
     5341 tgaccggcaa gttggacgcc cgcaagatcc gcgagattct cattaaggcc aagaagggcg
     5401 gcaagatcgc cgtgtaagaa ttcgcttatg cgtgcggccg cctcctttaa gaaggagata
     5461 tagatctctg taggtcccaa acaagctagt cttaatggag tcacattaat tggagaagcc
     5521 gtaaaaacac agttcaatta ttataagaaa gttgatggtg ttgtccaaca attacctgaa
     5581 acttacttta ctcagagtag aaatttacaa gaatttaaac ccaggagtca aatggaaatt
     5641 gatttcttag aattagctat ggatgaattc attgaacggt ataaattaga aggctatgcc
     5701 ttcgaacata tcgtttatgg agattttagt catagtcagt taggtggttt acatctactg
     5761 attggactag ctaaacgttt taaggaatca ccttttgaat tagaagattt tattcctatg
     5821 gacagtacag ttaaaaacta tttcataaca gatgcgcaaa caggttcatc taagtgtgtg
     5881 tgttctgtta ttgatttatt acttgatgat tttgttgaaa taataaaatc ccaagattta
     5941 tctgtagttt ctaaggttgt caaagtgact attgactata cagaaatttc atttatgctt
     6001 tggtgtaaag atggccatgt agaaacattt tacccaaaat tacaatctag tcaagcgtgg
     6061 caaccgggtg ttgctatgcc taatctttac aaaatgcaaa gaatgctatt agaaaagtgt
     6121 gaccttcaaa attatggtga tagtgcaaca ttacctaaag gcataatgat gaatgtcgca
     6181 aaatatactc aactgtgtca atatttaaac acattaacat tagctgtacc ctataatatg
     6241 agagttatac attttggtgc tggttctgat aaaggagttg caccaggtac agctgtttta
     6301 agacagtggt tgcctacggg tacgctgctt gtcgattcag atcttaatga ctttgtctct
     6361 gatgcagatt caactttgat tggtgattgt gcaactgtac atacagctaa taaatgggat
     6421 ctcattatta gtgatatgta cgaccctaag actaaaaatg ttacaaaaga aaatgactct
     6481 aaagagggtt ttttcactta catttgtggg tttatacaac aaaagctagc tcttggaggt
     6541 tccgtggcta taaagataga tcccaactcc ataagtgtgt cgacatataa ggatccactc
     6601 gacgactcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     6661 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     6721 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     6781 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     6841 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     6901 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     6961 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     7021 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     7081 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     7141 ttccgcatct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     7201 ctggaattaa ttcgagctcg gtaccttgat tctcgagtct agagggcccg tttaaacccg
     7261 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     7321 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     7381 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     7441 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     7501 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     7561 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     7621 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     7681 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     7741 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     7801 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     7861 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     7921 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     7981 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     8041 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     8101 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     8161 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     8221 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     8281 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     8341 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     8401 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     8461 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     8521 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     8581 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     8641 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     8701 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     8761 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     8821 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     8881 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     8941 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     9001 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     9061 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     9121 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     9181 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     9241 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     9301 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     9361 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     9421 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     9481 gtataccctc cacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     9541 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     9601 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     9661 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     9721 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     9781 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     9841 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     9901 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     9961 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
    10021 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
    10081 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
    10141 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
    10201 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
    10261 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
    10321 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
    10381 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
    10441 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
    10501 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
    10561 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
    10621 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
    10681 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
    10741 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
    10801 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
    10861 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
    10921 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
    10981 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
    11041 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
    11101 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
    11161 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
    11221 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
    11281 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
    11341 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
    11401 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
    11461 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
    11521 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
    11581 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
    11641 ttccgcgcac atttccccga aaagtgccac ctgacctcca cggatcggga gatctcccga
    11701 tcccctatgg tgcactctca gtacaatc
//
