LOCUS       sequence-462177-       10183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For packaging EGFP 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 10183)
  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 10183)
  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.4 EGFP-IRES-Luc-PS9
FEATURES             Location/Qualifiers
     source          1..10183
                     /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..1620)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKG"
     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"
     misc_feature    1731..2193
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1787..1804)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2014..2033
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      2214..2223
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2220..3872
                     /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    5067..5655
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5120..5140)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5538..5549)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5724..5741)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5730..5954
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6000..6428
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6087..6106)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6297..6318
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6437..6457)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6442..6771
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6622..6757
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6684..6703
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6838..7632
                     /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(6892..6911)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7502..7521
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7806..7927
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7843..7862)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7897..7916
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7976..7992)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7976..7992)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7989..8011)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8000..8016
                     /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(8024..8030,8031..8048,8049..8054))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8069..8090
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8207..8224)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8378..8966)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8458..8477)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9137..9928,9929..9997))
                     /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     9760..9779
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9998..10102)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(10177..10183,1..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 caagggataa
     1621 aatcaagaat tcgcttatgc taacgttact ggccgaagcc gcttggaata aggccggtgt
     1681 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     1741 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     1801 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
     1861 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     1921 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     1981 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     2041 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc
     2101 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     2161 gacgtggttt tcctttgaaa aacacgatga taaaacacac acacaccggt cgcgccgcca
     2221 tggaagatgc caaaaacatt aagaagggcc cagcgccatt ctacccactc gaagacggga
     2281 ccgccggcga gcagctgcac aaagccatga agcgctacgc cctggtgccc ggcaccatcg
     2341 cctttaccga cgcacatatc gaggtggaca ttacctacgc cgagtacttc gagatgagcg
     2401 ttcggctggc agaagctatg aagcgctatg ggctgaatac aaaccatcgg atcgtggtgt
     2461 gcagcgagaa tagcttgcag ttcttcatgc ccgtgttggg tgccctgttc atcggtgtgg
     2521 ctgtggcccc agctaacgac atctacaacg agcgcgagct gctgaacagc atgggcatca
     2581 gccagcccac cgtcgtattc gtgagcaaga aagggctgca aaagatcctc aacgtgcaaa
     2641 agaagctacc gatcatacaa aagatcatca tcatggatag caagaccgac taccagggct
     2701 tccaaagcat gtacaccttc gtgacttccc atttgccacc cggcttcaac gagtacgact
     2761 tcgtgcccga gagcttcgac cgggacaaaa ccatcgccct gatcatgaac agtagtggca
     2821 gtaccggatt gcccaagggc gtagccctac cgcaccgcac cgcttgtgtc cgattcagtc
     2881 atgcccgcga ccccatcttc ggcaaccaga tcatccccga caccgctatc ctcagcgtgg
     2941 tgccatttca ccacggcttc ggcatgttca ccacgctggg ctacttgatc tgcggctttc
     3001 gggtcgtgct catgtaccgc ttcgaggagg agctattctt gcgcagcttg caagactata
     3061 agattcaatc tgccctgctg gtgcccacac tatttagctt cttcgctaag agcactctca
     3121 tcgacaagta cgacctaagc aacttgcacg agatcgccag cggcggggcg ccgctcagca
     3181 aggaggtagg tgaggccgtg gccaaacgct tccacctacc aggcatccgc cagggctacg
     3241 gcctgacaga aacaaccagc gccattctga tcacccccga aggggacgac aagcctggcg
     3301 cagtaggcaa ggtggtgccc ttcttcgagg ctaaggtggt ggacttggac accggtaaga
     3361 cactgggtgt gaaccagcgc ggcgagctgt gcgtccgtgg ccccatgatc atgagcggct
     3421 acgttaacaa ccccgaggct acaaacgctc tcatcgacaa ggacggctgg ctgcacagcg
     3481 gcgacatcgc ctactgggac gaggacgagc acttcttcat cgtggaccgg ctgaagagcc
     3541 tgatcaaata caagggctac caggtagccc cagccgaact ggagagcatc ctgctgcaac
     3601 accccaacat cttcgacgcc ggggtcgccg gcctgcccga cgacgatgcc ggcgagctgc
     3661 ccgccgcagt cgtcgtgctg gaacacggta aaaccatgac cgagaaggag atcgtggact
     3721 atgtggccag ccaggttaca accgccaaga agctgcgcgg tggtgttgtg ttcgtggacg
     3781 aggtgcctaa aggactgacc ggcaagttgg acgcccgcaa gatccgcgag attctcatta
     3841 aggccaagaa gggcggcaag atcgccgtgt aagaattcgc ttatgcgtgc ggccgcctcc
     3901 tttaagaagg agatatagat ctctgtaggt cccaaacaag ctagtcttaa tggagtcaca
     3961 ttaattggag aagccgtaaa aacacagttc aattattata agaaagttga tggtgttgtc
     4021 caacaattac ctgaaactta ctttactcag agtagaaatt tacaagaatt taaacccagg
     4081 agtcaaatgg aaattgattt cttagaatta gctatggatg aattcattga acggtataaa
     4141 ttagaaggct atgccttcga acatatcgtt tatggagatt ttagtcatag tcagttaggt
     4201 ggtttacatc tactgattgg actagctaaa cgttttaagg aatcaccttt tgaattagaa
     4261 gattttattc ctatggacag tacagttaaa aactatttca taacagatgc gcaaacaggt
     4321 tcatctaagt gtgtgtgttc tgttattgat ttattacttg atgattttgt tgaaataata
     4381 aaatcccaag atttatctgt agtttctaag gttgtcaaag tgactattga ctatacagaa
     4441 atttcattta tgctttggtg taaagatggc catgtagaaa cattttaccc aaaattacaa
     4501 tctagtcaag cgtggcaacc gggtgttgct atgcctaatc tttacaaaat gcaaagaatg
     4561 ctattagaaa agtgtgacct tcaaaattat ggtgatagtg caacattacc taaaggcata
     4621 atgatgaatg tcgcaaaata tactcaactg tgtcaatatt taaacacatt aacattagct
     4681 gtaccctata atatgagagt tatacatttt ggtgctggtt ctgataaagg agttgcacca
     4741 ggtacagctg ttttaagaca gtggttgcct acgggtacgc tgcttgtcga ttcagatctt
     4801 aatgactttg tctctgatgc agattcaact ttgattggtg attgtgcaac tgtacataca
     4861 gctaataaat gggatctcat tattagtgat atgtacgacc ctaagactaa aaatgttaca
     4921 aaagaaaatg actctaaaga gggttttttc acttacattt gtgggtttat acaacaaaag
     4981 ctagctcttg gaggttccgt ggctataaag atagatccca actccataag tgtgtcgaca
     5041 tataaggatc cactcgacga ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     5101 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     5161 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     5221 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     5281 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     5341 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     5401 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     5461 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     5521 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     5581 cggctctgcg gcctcttccg catcttcgcc ttcgccctca gacgagtcgg atctcccttt
     5641 gggccgcctc cccgcctgga attaattcga gctcggtacc ttgattctcg agtctagagg
     5701 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     5761 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     5821 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     5881 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     5941 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     6001 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     6061 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     6121 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     6181 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     6241 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     6301 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     6361 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     6421 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     6481 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     6541 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     6601 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     6661 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     6721 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     6781 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     6841 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     6901 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     6961 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     7021 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     7081 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     7141 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     7201 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     7261 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     7321 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     7381 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     7441 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     7501 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     7561 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     7621 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     7681 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     7741 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     7801 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     7861 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     7921 tatcttatca tgtctgtata ccctccacct ctagctagag cttggcgtaa tcatggtcat
     7981 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     8041 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     8101 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     8161 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     8221 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     8281 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     8341 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     8401 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     8461 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     8521 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     8581 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     8641 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     8701 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     8761 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     8821 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     8881 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     8941 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     9001 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     9061 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     9121 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     9181 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     9241 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     9301 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     9361 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     9421 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     9481 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     9541 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     9601 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     9661 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     9721 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     9781 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     9841 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     9901 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     9961 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
    10021 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
    10081 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac ctccacggat
    10141 cgggagatct cccgatcccc tatggtgcac tctcagtaca atc
//
