Luc-T20

LOCUS      Luc-T20               9866 bp ds-DNA     circular SYN 17-NOV-2021
DEFINITION  Transient mammalian expression of Luc2 (firefly luciferase) along 
            with SARS-CoV-2 sequence 20080-22222 (T20) within the 3'UTR. 
            Resulting transcript is packaged into SARS-CoV-2 virus-like 
            particles..
ACCESSION   .
VERSION     .
KEYWORDS    Luc-T20
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9866)
  AUTHORS   Syed AM, Taha TY, Tabata T, Chen IP, Ciling A, Khalid MM, Sreekumar 
            B, Chen PY, Hayashi JM, Soczek KM, Ott M, Doudna JA
  TITLE     Rapid assessment of SARS-CoV-2 evolved variants using virus-like 
            particles.
  JOURNAL   Science. 2021 Nov 4:eabl6184. doi: 10.1126/science.abl6184.
  PUBMED    34735219
REFERENCE   2  (bases 1 to 9866)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 17, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..9866
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        137..516
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        517..720
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     671..691
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     765..784
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        765..783
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      803..812
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             809..2461
                     /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    4702..5290
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4755..4775)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5173..5184)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5359..5376)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5365..5589
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5635..6063
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5722..5741)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5932..5953
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6072..6092)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6077..6406
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6257..6392
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6319..6338
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6473..7267
                     /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(6527..6546)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7137..7156
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7441..7562
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7478..7497)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7532..7551
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7611..7627)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7611..7627)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7624..7646)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7635..7651
                     /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(7659..7689)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7704..7725
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7842..7859)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8013..8601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8093..8112)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8772..9632)
                     /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     9395..9414
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9633..9737)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9812..9831)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
       61 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
      121 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
      181 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      241 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      301 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      361 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      421 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      481 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      541 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      601 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      661 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      721 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat
      781 agggagaccc aagctggcta gcgccgccat ggaagatgcc aaaaacatta agaagggccc
      841 agcgccattc tacccactcg aagacgggac cgccggcgag cagctgcaca aagccatgaa
      901 gcgctacgcc ctggtgcccg gcaccatcgc ctttaccgac gcacatatcg aggtggacat
      961 tacctacgcc gagtacttcg agatgagcgt tcggctggca gaagctatga agcgctatgg
     1021 gctgaataca aaccatcgga tcgtggtgtg cagcgagaat agcttgcagt tcttcatgcc
     1081 cgtgttgggt gccctgttca tcggtgtggc tgtggcccca gctaacgaca tctacaacga
     1141 gcgcgagctg ctgaacagca tgggcatcag ccagcccacc gtcgtattcg tgagcaagaa
     1201 agggctgcaa aagatcctca acgtgcaaaa gaagctaccg atcatacaaa agatcatcat
     1261 catggatagc aagaccgact accagggctt ccaaagcatg tacaccttcg tgacttccca
     1321 tttgccaccc ggcttcaacg agtacgactt cgtgcccgag agcttcgacc gggacaaaac
     1381 catcgccctg atcatgaaca gtagtggcag taccggattg cccaagggcg tagccctacc
     1441 gcaccgcacc gcttgtgtcc gattcagtca tgcccgcgac cccatcttcg gcaaccagat
     1501 catccccgac accgctatcc tcagcgtggt gccatttcac cacggcttcg gcatgttcac
     1561 cacgctgggc tacttgatct gcggctttcg ggtcgtgctc atgtaccgct tcgaggagga
     1621 gctattcttg cgcagcttgc aagactataa gattcaatct gccctgctgg tgcccacact
     1681 atttagcttc ttcgctaaga gcactctcat cgacaagtac gacctaagca acttgcacga
     1741 gatcgccagc ggcggggcgc cgctcagcaa ggaggtaggt gaggccgtgg ccaaacgctt
     1801 ccacctacca ggcatccgcc agggctacgg cctgacagaa acaaccagcg ccattctgat
     1861 cacccccgaa ggggacgaca agcctggcgc agtaggcaag gtggtgccct tcttcgaggc
     1921 taaggtggtg gacttggaca ccggtaagac actgggtgtg aaccagcgcg gcgagctgtg
     1981 cgtccgtggc cccatgatca tgagcggcta cgttaacaac cccgaggcta caaacgctct
     2041 catcgacaag gacggctggc tgcacagcgg cgacatcgcc tactgggacg aggacgagca
     2101 cttcttcatc gtggaccggc tgaagagcct gatcaaatac aagggctacc aggtagcccc
     2161 agccgaactg gagagcatcc tgctgcaaca ccccaacatc ttcgacgccg gggtcgccgg
     2221 cctgcccgac gacgatgccg gcgagctgcc cgccgcagtc gtcgtgctgg aacacggtaa
     2281 aaccatgacc gagaaggaga tcgtggacta tgtggccagc caggttacaa ccgccaagaa
     2341 gctgcgcggt ggtgttgtgt tcgtggacga ggtgcctaaa ggactgaccg gcaagttgga
     2401 cgcccgcaag atccgcgaga ttctcattaa ggccaagaag ggcggcaaga tcgccgtgta
     2461 agaattcgct tatgcggccg cctcctttaa gaaggagata tagatctctg taggtcccaa
     2521 acaagctagt cttaatggag tcacattaat tggagaagcc gtaaaaacac agttcaatta
     2581 ttataagaaa gttgatggtg ttgtccaaca attacctgaa acttacttta ctcagagtag
     2641 aaatttacaa gaatttaaac ccaggagtca aatggaaatt gatttcttag aattagctat
     2701 ggatgaattc attgaacggt ataaattaga aggctatgcc ttcgaacata tcgtttatgg
     2761 agattttagt catagtcagt taggtggttt acatctactg attggactag ctaaacgttt
     2821 taaggaatca ccttttgaat tagaagattt tattcctatg gacagtacag ttaaaaacta
     2881 tttcataaca gatgcgcaaa caggttcatc taagtgtgtg tgttctgtta ttgatttatt
     2941 acttgatgat tttgttgaaa taataaaatc ccaagattta tctgtagttt ctaaggttgt
     3001 caaagtgact attgactata cagaaatttc atttatgctt tggtgtaaag atggccatgt
     3061 agaaacattt tacccaaaat tacaatctag tcaagcgtgg caaccgggtg ttgctatgcc
     3121 taatctttac aaaatgcaaa gaatgctatt agaaaagtgt gaccttcaaa attatggtga
     3181 tagtgcaaca ttacctaaag gcataatgat gaatgtcgca aaatatactc aactgtgtca
     3241 atatttaaac acattaacat tagctgtacc ctataatatg agagttatac attttggtgc
     3301 tggttctgat aaaggagttg caccaggtac agctgtttta agacagtggt tgcctacggg
     3361 tacgctgctt gtcgattcag atcttaatga ctttgtctct gatgcagatt caactttgat
     3421 tggtgattgt gcaactgtac atacagctaa taaatgggat ctcattatta gtgatatgta
     3481 cgaccctaag actaaaaatg ttacaaaaga aaatgactct aaagagggtt ttttcactta
     3541 catttgtggg tttatacaac aaaagctagc tcttggaggt tccgtggcta taaagataac
     3601 agaacattct tggaatgctg atctttataa gctcatggga cacttcgcat ggtggacagc
     3661 ctttgttact aatgtgaatg cgtcatcatc tgaagcattt ttaattggat gtaattatct
     3721 tggcaaacca cgcgaacaaa tagatggtta tgtcatgcat gcaaattaca tattttggag
     3781 gaatacaaat ccaattcagt tgtcttccta ttctttattt gacatgagta aatttcccct
     3841 taaattaagg ggtactgctg ttatgtcttt aaaagaaggt caaatcaatg atatgatttt
     3901 atctcttctt agtaaaggta gacttataat tagagaaaac aacagagttg ttatttctag
     3961 tgatgttctt gttaacaact aaacgaacaa tgtttgtttt tcttgtttta ttgccactag
     4021 tctctagtca gtgtgttaat cttacaacca gaactcaatt accccctgca tacactaatt
     4081 ctttcacacg tggtgtttat taccctgaca aagttttcag atcctcagtt ttacattcaa
     4141 ctcaggactt gttcttacct ttcttttcca atgttacttg gttccatgct atacatgtct
     4201 ctgggaccaa gggtactaag aggtttgata accctgtcct accatttaat gatggtgttt
     4261 attttgcttc cactgagaag tctaacataa taagaggctg gatttttggt actactttag
     4321 attcgaagac ccagtcccta cttattgtta ataacgctac taatgttgtt attaaagtct
     4381 gtgaatttca attttgtaat gatccatttt tgggtgttta ttaccacaaa aacaacaaaa
     4441 gttggatgga aagtgagttc agagtttatt ctagtgcgaa taattgcact tttgaatatg
     4501 tctctcagcc ttttcttatg gaccttgaag gaaaacaggg taatttcaaa aatcttaggg
     4561 aatttgtgtt taagaatatt gatggttatt ttaaaatata ttctaagcac acgcctatta
     4621 atttagtgcg tgatctccct cagggtttga tcccaactcc ataagtgtgt cgacatataa
     4681 ggatccactc gacgactcga caatcaacct ctggattaca aaatttgtga aagattgact
     4741 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     4801 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     4861 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     4921 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     4981 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     5041 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     5101 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     5161 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     5221 ctgcggcctc ttccgcatct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     5281 gcctccccgc ctggaattaa ttcgagctcg gtaccttgat tctcgagtct agagggcccg
     5341 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     5401 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     5461 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     5521 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     5581 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     5641 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5701 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5761 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5821 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     5881 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5941 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     6001 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     6061 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     6121 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     6181 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     6241 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     6301 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     6361 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     6421 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     6481 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     6541 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     6601 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     6661 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     6721 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     6781 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     6841 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     6901 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     6961 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     7021 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     7081 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     7141 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     7201 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     7261 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     7321 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     7381 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     7441 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     7501 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     7561 tatcatgtct gtataccctc cacctctagc tagagcttgg cgtaatcatg gtcatagctg
     7621 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     7681 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     7741 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     7801 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     7861 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     7921 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     7981 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     8041 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     8101 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     8161 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     8221 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     8281 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     8341 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     8401 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     8461 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     8521 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     8581 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     8641 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     8701 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     8761 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     8821 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     8881 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     8941 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     9001 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     9061 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     9121 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     9181 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     9241 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     9301 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     9361 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     9421 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     9481 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     9541 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     9601 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     9661 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     9721 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacctcca cggatcggga
     9781 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     9841 gccagtatct gctccctgct tgtgtg
//