LOCUS       sequence-442644-       14043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Competition assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14043)
  AUTHORS   Uijttewaal ECH, Lee J, Sell AC, Botay N, Vainorius G, Novatchkova M,
            Baar J, Yang J, Potzler T, van der Leij S, Lowden C, Sinner J, 
            Elewaut A, Gavrilovic M, Obenauf A, Schramek D, Elling U
  TITLE     CRISPR-StAR enables high-resolution genetic screening in complex in 
            vivo models.
  JOURNAL   Nat Biotechnol. 2024 Dec 16. doi: 10.1038/s41587-024-02512-9.
  PUBMED    39681701
REFERENCE   2  (bases 1 to 14043)
  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: "Nat 
            Biotechnol. 2024 Dec 16. doi: 10.1038/s41587-024-02512-9."
COMMENT     Sequence Label: pLenti-TripleColour-StAR
FEATURES             Location/Qualifiers
     source          1..14043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(190..981,982..1050))
                     /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     813..832
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1051..1155)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1230..1249)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1425..1804
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1806..2004
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1959..1979
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2022..2202
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2249..2374
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2867..3100
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3285..3329
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    3627..3744
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3872..4120
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3872..3892
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4043..4062
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    complement(4164..4197)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4525..5704
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4758..5695
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5653..5673
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             5739..6437
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     protein_bind    6438..6471
                     /label=lox5171
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTGTAC) (Shaw et al., 2021). lox5171 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             6474..6521
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6522..6578
                     /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             6579..7523
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 703 nm (Shcherbakova et al., 2016)"
                     /label=miRFP703
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSLVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPIHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     CDS             7524..7577
                     /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             7572..8372
                     /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="GPMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWAGRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7632..7651)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8242..8261
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    complement(8382..8415)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        8499..8998
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8534..8557)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     8931..8950
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             9012..9713
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MLSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(9145..9163)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     9413..9432
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     9607..9630
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    9723..9756
                     /label=lox5171
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTGTAC) (Shaw et al., 2021). lox5171 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     misc_feature    9918..10506
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9971..9991)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10389..10400)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10509..10525)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10510..10526)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             11031..11211
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11237..11254)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11243..11467
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11513..11941
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11600..11619)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11810..11831
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11950..11970)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11955..12284
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12135..12270
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12197..12216
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12332..12379
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12398..12772
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    12902..13023
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12939..12958)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12993..13012
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13072..13088)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13072..13088)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13085..13107)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13096..13112
                     /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(13120..13126,13127..13144,13145..13150))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13165..13186
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13303..13320)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(13474..14043,1..19))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13554..13573)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
       61 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
      121 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
      181 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
      241 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
      301 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
      361 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
      421 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
      481 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
      541 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
      601 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
      661 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
      721 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
      781 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
      841 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
      901 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
      961 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     1021 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     1081 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     1141 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     1201 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     1261 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     1321 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
     1381 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgcg cgttgacatt gattattgac
     1441 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     1501 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     1561 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     1621 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     1681 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     1741 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     1801 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     1861 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     1921 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     1981 acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc
     2041 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     2101 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     2161 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     2221 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
     2281 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
     2341 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     2401 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     2461 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
     2521 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     2581 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     2641 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
     2701 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     2761 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     2821 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     2881 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     2941 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     3001 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     3061 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     3121 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     3181 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     3241 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     3301 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     3361 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     3421 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     3481 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
     3541 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc
     3601 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     3661 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     3721 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acaggctagc ctctaggcgc
     3781 cggaattgaa gatctccgcg gttaattaaa atgatacggc gaccaccgag atctgtcttg
     3841 atctaggagc tactcatcaa cggtcacatg tgagggccta tttcccatga ttccttcata
     3901 tttgcatata cgatacaagg ctgttagaga gataattgga attaatttga ctgtaaacac
     3961 aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt
     4021 tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga
     4081 tttcttggct ttatatatct tgtggaaagg acgaaacacc cgagacgtct accgcacgtc
     4141 gtctcagttt aagagctatg ctgataactt cgtatagcat acattatacg aagttatttt
     4201 ttttttttgc tcgagtagaa gttaacggtc atcattgaat ctggaggaga gctaagttaa
     4261 taaggtcggt gatggttggt tagatttatg tcgtagtcac atccacgcgg ccgctgtatt
     4321 cacgactgcg tgtctgaact atcatctggg actgcttaat cggatgtgtc tcatccaact
     4381 gagttggggg ctactgagag cagaaatagc aagtaactca ttaggtaaat aaggtagtcc
     4441 gtttagtgcc gtagccaatt ctcagacatt taatggctct aaacgtaaca tttacaaatt
     4501 ttcgaagctc cggtgcccgt cagtggctcc ggtgcccgtc agtgggcaga gcgcacatcg
     4561 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt cggctccggt gcctagagaa
     4621 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     4681 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     4741 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
     4801 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
     4861 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
     4921 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
     4981 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
     5041 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
     5101 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
     5161 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
     5221 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
     5281 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
     5341 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
     5401 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     5461 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     5521 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     5581 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     5641 cttggttcat tctcaagcct cagacagtgg ttcaaagttt tttcttccat ttcaggtgtc
     5701 gtgacgtacg gccaccatga gcggcctctg aggccaccat gagcgagctg attaaggaga
     5761 acatgcacat gaagctgtac atggagggca ccgtggacaa ccatcacttc aagtgcacat
     5821 ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag gtggtcgagg
     5881 gcggccctct ccccttcgcc ttcgacatcc tggctactag cttcctctac ggcagcaaga
     5941 ccttcatcaa ccacacccag ggcatccccg acttcttcaa gcagtccttc cctgagggct
     6001 tcacatggga gagagtcacc acatacgagg acgggggcgt gctgaccgct acccaggaca
     6061 ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcacat
     6121 ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggccttcacc gaaacgctgt
     6181 accccgctga cggcggcctg gaaggcagaa acgacatggc cctgaagctc gtgggcggga
     6241 gccatctgat cgcaaacatc aagaccacat atagatccaa gaaacccgct aagaacctca
     6301 agatgcctgg cgtctactat gtggactaca gactggaaag aatcaaggag gccaacaacg
     6361 agacctacgt cgagcagcac gaggtggcag tggccagata ctgcgacctc cctagcaaac
     6421 tggggcacaa gcttaatata acttcgtata atgtgtacta tacgaagtta tctaagcgac
     6481 ctgccgccac aaagaaggct ggacaggcta agaagaagaa agccaccaac ttcagcctgc
     6541 tgaagcaggc cggcgacgtg gaggagaacc ccggccccat ggtagcaggt catgcctctg
     6601 gcagccccgc attcgggacc gcctctcatt cgaattgcga acatgaagag atccacctcg
     6661 ccggctcgat ccagccgcat ggcgcgcttc tggtcgtcag cgaacatgat catcgcgtca
     6721 tccaggccag cgccaacgcc gcggaatttc tgaatctcgg aagcgtactc ggcgttccgc
     6781 tcgccgagat cgacggcgat ctgttgatca agatcctgcc gcatctcgat cccaccgccg
     6841 aaggcatgcc ggtcgcggtg cgctgccgga tcggcaatcc ctctacggag tactgcggtc
     6901 tgatgcatcg gcctccggaa ggcgggctga tcatcgaact cgaacgtgcc ggcccgtcga
     6961 tcgatctgtc aggcacgctg gcgccggcgc tggagcggat ccgcacggcg ggttcactgc
     7021 gcgcgctgtg cgatgacacc gtgctgctgt ttcagcagtg caccggctac gaccgggtga
     7081 tggtgtatcg tttcgatgag caaggccacg gcctggtatt ctccgagtgc catgtgcctg
     7141 ggctcgaatc ctatttcggc aaccgctatc cgtcgtcgct ggtcccgcag atggcgcggc
     7201 agctgtacgt gcggcagcgc gtccgcgtgc tggtcgacgt cacctatcag ccggtgccgc
     7261 tggagccgcg gctgtcgccg ctgaccgggc gcgatctcga catgtcgggc tgcttcctgc
     7321 gctcgatgtc gccgatccat ctgcagttcc tgaaggacat gggcgtgcgc gccaccctgg
     7381 cggtgtcgct ggtggtcggc ggcaagctgt ggggcctggt tgtctgtcac cattatctgc
     7441 cgcgcttcat ccgtttcgag ctgcgggcga tctgcaaacg gctcgccgaa aggatcgcga
     7501 cgcggatcac cgcgcttgag agcgagggca gaggcagtct gctgacatgc ggtgacgtgg
     7561 aagagaatcc cggccctatg attgaacaag atggattgca cgcaggttct ccggccgctt
     7621 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     7681 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     7741 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     7801 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     7861 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     7921 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     7981 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     8041 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     8101 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     8161 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     8221 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     8281 aatgggctgg ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     8341 ccttctatcg ccttcttgac gagttcttct gaacctgcag gataacttcg tatagcatac
     8401 attatacgaa gttatcagca tagcaagttt aaataaggct agtccgttat caacttgaaa
     8461 aagtggcacc gagtcggtgc ttttttgttt tagttaacgg gtaggggagg cgcttttccc
     8521 aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct acacaagtgg
     8581 cctctggcct cgcacacatt ccacatccac cggtaggcgc caaccggctc cgttctttgg
     8641 tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc ccgccccgca
     8701 gctcgcgtcg tgcaggacgt gacaaatgga agtagcacct ctcactagtc tcgtgcagat
     8761 ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc aatagcagct
     8821 ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg gcgggctcag
     8881 gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac
     8941 gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac
     9001 ctgcagccaa tatgttgagc aagggcgagg aggtcatcaa agagttcatg cgcttcaagg
     9061 tgcgcatgga gggctccatg aacggccatg agttcgagat cgagggcgag ggcgagggcc
     9121 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct
     9181 tcgcctggga catcctgtcc ccccagttca tgtacggctc caaggcgtac gtgaagcacc
     9241 ccgccgacat ccccgattac aagaagctgt ccttccccga gggcttcaag tgggagcgcg
     9301 tgatgaactt cgaggacggc ggtctggtga ccgtgaccca ggactcctcc ctccaggacg
     9361 gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt cccccccgac ggccccgtaa
     9421 tgcagaagaa gacaatgggc tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg
     9481 tgctgaaggg cgagatccac caggccctga agctgaagga cggcggccac tacctggtgg
     9541 agttcaagac catctacatg gccaagaagc ccgtgcaact gcccggctac tactacgtgg
     9601 acaccaagct ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc
     9661 gctccgaggg ccgccaccac ctgttcctgt acggcatgga cgagctgtac aagtgaggat
     9721 ccataacttc gtataatgtg tactatacga agttatagtc tagaggtacc cgttaagatc
     9781 ggaagagcac acgtctgaac tccagtcaca aaattatctc gtatgccgtc ttatgcttgt
     9841 taattaacaa ttgacgcgta attctaccgg ctagcgagag cacacacgaa aggtaattcg
     9901 atatcaagct tatcggtaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     9961 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
    10021 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
    10081 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
    10141 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
    10201 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
    10261 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
    10321 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
    10381 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
    10441 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
    10501 ccccgcatcg ataccgtcga cctcgagacc tagaaaaaca tggagcaatc acaagtagca
    10561 atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg
    10621 gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc
    10681 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
    10741 aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat tggcagaact
    10801 acacaccagg gccagggatc agatatccac tgacctttgg atggtgctac aagctagtac
    10861 cagttgagca agagaaggta gaagaagcca atgaaggaga gaacacccgc ttgttacacc
    10921 ctgtgagcct gcatgggatg gatgacccgg agagagaagt attagagtgg aggtttgaca
    10981 gccgcctagc atttcatcac atggcccgag agctgcatcc ggactgtact gggtctctct
    11041 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
    11101 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
    11161 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agggcccgtt
    11221 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
    11281 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
    11341 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
    11401 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
    11461 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
    11521 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
    11581 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
    11641 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
    11701 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
    11761 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
    11821 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
    11881 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
    11941 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
    12001 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
    12061 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
    12121 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
    12181 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
    12241 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
    12301 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat
    12361 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
    12421 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
    12481 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
    12541 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
    12601 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
    12661 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
    12721 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct
    12781 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
    12841 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
    12901 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
    12961 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
    13021 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
    13081 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
    13141 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
    13201 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
    13261 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
    13321 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
    13381 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
    13441 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
    13501 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
    13561 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
    13621 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
    13681 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
    13741 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
    13801 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
    13861 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
    13921 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
    13981 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
    14041 gcg
//
